High Expression of PYK2 is Associated with Poor Prognosis and Cancer Progression in Early- Stage Cervical Carcinoma

High Expression of PYK2 is Associated with Poor Prognosis and Cancer Progression in Early- Stage Cervical Carcinoma | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article High Expression of PYK2 is Associated with Poor Prognosis and Cancer Progression in Early- Stage Cervical Carcinoma Can Zhang, Xinghua Zhu, Yong Li, Jia Shao, Haibo Xu, Lei Chen, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-696303/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Proline-rich tyrosine kinase-2 (PYK2), also known as calcium dependent tyrosine kinase (CADTK), regulates different signal transduction cascades that control cell proliferation, migration and invasion. However, the role of PYK2 in cervical cancer remains to be elucidated. Methods: PYK2 expression was detected in tissue microarray and cervical cancer cell lines. Statistical analysis was performed to evaluate its clinicopathological significance. Small interfering RNA (SiRNA) was employed to suppress endogenous PYK2 expression in cervical cancer cells to observe the biological function. Results: PYK2 expression was up-regulated in cervical cancer specimens compared with paired adjacent normal cervical tissue samples. Statistical analyses indicated that PYK2 expression might be an independent prognostic indicator for patients with early-stage cervical cancer. A nomogram model was constructed based on PYK2 expression and other clinicopathological risk factors, and it performed well in predicting patients survival. In cellular studies, down-regulation of PYK2 remarkably inhibited cellular proliferation, migration and invasion. Conclusion: PYK2 expression possessed the potential to serve as a novel prognostic marker in cervical cancer patients. Surgery Oncology PYK2 early-stage cervical carcinoma prognosis nomogram biomaker Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Cervical cancer is the most frequent gynecological cancer and the fourth leading cause of globally cancer-related motility among women, especially in developing countries[ 1 ].As known, persistent infection of high-risk oncogenic types of human papillomavirus (HPV) is the primary cause of cervical cancer. However, only a small percentage of patients infected develop into invasive cancer, indicating the carcinogenic effects of other factors[ 2 ]. Although clinical factors, such as stage, squamous cell carcinoma antigen (SCC), lymph node metastasis, may serve as prognostic markers, more clinical trials are needed to confirm and validate the clinical sensitivity[ 3 – 5 ]. Therefore, it is crucial to obtain a better understanding of the underlying molecular mechanism of cervical cancer development and progression, and novel biomarkers are required to investigate, which would provide reliable predictors as well as potential targets for cervical cancer. Proline-rich tyrosine kinase-2 (PYK2) is a non-receptor tyrosine kinase, which belongs to the focal adhesion kinase (FAK) family and regulates cell adhesion, morphology, migration, and invasion via different signal transduction cascades[ 6 – 9 ]. PYK2 consists of three domains associated with tumor progression and can be activated by multiple growth factors, neuropeptides, cytokines, hormones, and chemokines[ 10 – 15 ]. Overexpression of PYK2 has been reported to promote progression and lead to poor prognosis in many kinds of malignant tumors such as breast cancer, liver cancer, lung cancer and pancreatic cancer[ 7 , 16 – 22 ]. PYK2 was only found to serve as an onco-suppressor gene in androgen-dependent prostate cancer cells and neuroblastoma cells so far[ 23 , 24 ]. However, the role of PYK2 in cervical cancer is still unclear. In our study, we evaluated the role of Pyk2 in early-stage cervical carcinoma and found that the high expression of Pyk2 is associated with poor clinical outcomes in cervical cancer patients. Furthermore, we gained insight into the important functions of this protein in cervical cancer lines. Materials And Methods Patients and clinical samples The study was composed of 268 samples from 134 cervical cancer patients, who treated with radical hysterectomy plus pelvic lymphadenectomy without neoadjuvant chemotherapy or preoperative radiotherapy at Affiliated Tumor Hospital of Nantong University from December 2007 to September 2014. Tissue microarray (TMA) was prepared with the 134 cervical cancer tissues and adjacent normal cervical tissue samples. The study was performed with the approval of the Ethics Committee of Affiliated Tumor Hospital of Nantong University and complied with the Helsinki Declaration. Cell culture and transfection HeLa was obtained from Medical College of Nantong University and SiHa was purchased from the Cell bank of the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). SiHa cells were cultured in minimum essential medium ( HyClone Laboratories, Logan, UT) and Hela cells were cultured in DMEM medium(Biological Industries, Israel), which were all supplemented with 10 % fetal bovine serum (Biological Industries, Israel) and 1 % antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin) in a 5% CO2-humidified atmosphere at 37℃. To silence endogenous PYK2 expression, the following Pyk2 small interfering RNA (siRNA) were synthesized by GenePharma (Shanghai, China) with the sequence of 5'-GAUGUGGUCCUGAAUCGUATT-3'. A non-silencing siRNA oligonucleotide that does not recognize any known mammalian gene homolog (GenePharma) was used as a negative control. SiRNA-knockdown were performed with LipoMax™ transfection reagent (SUDGEN) following the manufacturer’s procedure. The transfection efficiencies were tested by Western blot analysis. Immunohistochemistry (IHC) stainingand evaluation The expression of PYK2 was determined using immunohistochemistry (IHC) staining of tissue microarrays (TMAs). A standard immunostaining procedure was performed using Rabbit molyclonal anti-Pyk2 antibody (1:100 dilution; ab32571, Abcam, Cambridge, MA, USA) at 4℃ overnight. Two independent pathologists blind to the clinical or pathologic data assessed the IHC score. The staining intensity was scored according to 4 grades: 0 (No staining), 1 (weak), 2 (moderate), or 3 (intense). The percentage of tumor cells stained was scored as follows: 1, 1–25%; 2, 26–50%; 3, 51–75%; or 4, 76–100%. The overall IHC score was finally obtained by multiplying the intensity score and percentage scores (ranged from 0 to 12). The samples were considered to be high expression if the score was > 4, and defined as low expression if not. Western Blotting Samples from cell lysates or tissues lysates were prepared using cold RIPA buffer and separated by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide). Then proteins were transferred onto PVDF (polyvinylidene difluoride) membranes and incubated with primary antibodies including PYK2 (1:2000 dilution; ab32571, Abcam, Cambridge, MA, USA), anti-GAPDH(1:5000 dilution; Santa Cruz, CA, USA). Cell proliferation assay For CCK8 assay, HeLa and SiHa cells cells were seeded into 96-well plates at density of 3000 cells per well. At the specified time point, 10µl CCK8 solution was added and incubated for 2hours. Cell viability was determined using enzyme-labeled instrument to measure absorbance at 450 nm. Transwell assay A total of 2×10 4 HeLa and SiHa cells were seeded into the upper part of a transwell chamber with a pore size of 8 µm (BD, Franklin Lakes, NJ, USA). For the invasion assay, the upper chamber was coated with 60µl prechilled Matrigel. The bottom chambers were filled with 0.5 mL of medium supplemented with 10% FBS and incubated for 24h. The transmembraned cells were fixed with 100% methanol and stained with 0.1% crystal violet for 15min. Finally, the number of migrated and invading cells on the lower side of the membrane was counted under a light microscope. Wound healing assay Cells were inculated into 6-well plates and grown in complete medium until 90–100% confluency. A 5mm-diameter wound was made across the cells using a sterile plastic tip. The complete medium was replaced by serum-free medium then. The migration of the cells was observed under a microscope at 0h, 24h and 48h respectively. Statistical analysis The relationship between CPE expression and the clinicopathological characteristics was assessed using the Chi-squared test or Fisher’s exact test. Disease-free survival and overall survival were calculated using the Kaplan-Meier method and were compared using the log-rank test. Univariate and multivariate Cox proportional hazards regression models were performed to identify the independent prognostic factors. Results were reported as hazard ratio (HR) and 95% confidence interval (CI). A two-sided P < 0.05 was considered to indicate statistical significance. All statistical analysis were obtained by using IBM SPSS software (Statistical Package for the Social Sciences; Version 25.0). Nomograms and calibration plots for OS and DFS based on multivariable analysis were established with R software (version 3.6.1). Results PYK2 expression with clinicopathological features PYK2 was mainly localized in the cytoplasm of cervical cancer cells as shown in Fig. 1 . Negative or low PYK2 protein expression was detected in adjacent normal cervical tissue samples. Among the 134 cervical cancer specimens, 62 samples (46.3%) showed high PYK2 protein expression and low expression of PYK2 was observed in 72 tumor samples. The clinicopathologic analysis exhibited that PYK2 expression was strongly correlated with differentiation grade (P = 0.027). However, PYK2 expression did not correlate with other clinical features, including age, FIGO (International Federation of Gynecology and Obstetrics) stage, pathological type, tumor size, depth of invasion, lympho-vascular space invasion, lymph node metastasis and adjuvant therapy (Table 1 ). Table 1 Association between PYK2 expression and clinicopathological variables in cervical cancer patients Variables Total (n = 134) PYK2 P-value Low expression (%) High expression (%) Age ≤ 52 years 64 (47.8%) 31 (48.4%) 33 (51.6%) 0.240 > 52 years 70 (52.2%) 41 (58.6%) 29 (41.4%) FIGO stage IB 121 (90.3%) 65 (53.7%) 56 (46.3%) 0.993 IIA 13 (9.70%) 7(53.8%) 6 (46.2%) Pathological type SCC 115 (85.8%) 61 (53.0%) 54 (47.0%) 0.694 No-SCC 19 (14.2%) 11 (57.9%) 8 (42.1%) Tumor Size ≤ 2cm 33 (24.6%) 15(45.5%) 18 (54.5%) 0.272 > 2cm 101 (75.4%) 57 (56.4%) 44 (43.6%) Histologic grade I-II 110 (80.3%) 64 (58.2%) 46 (41.8%) 0.027* III 24 (17.9%) 8 (33.3%) 16 (66.7%) Depth of invasion < 2/3 99 (73.9%) 51 (51.5%) 48 (48.5%) 0.387 ≥ 2/3 35 (26.1%) 21 (60.0%) 14 (40.0%) LVSI No 113 (84.3%) 60 (53.1%) 53 (46.9%) 0.733 Yes 21(15.7%) 12 (57.1%) 9 (42.9%) LNM No 110 (82.1%) 57 (51.8%) 53(48.2%) 0.342 Yes 24 (17.9%) 15 (62.5%) 9 (37.5%) Adjuvant therapy No 49 (36.6%) 23(46.9%) 26 (53.1%) 0.278 Chemoradiotherapy 42 (31.3%) 21 (50.0%) 21 (50.0%) Chemotherapy 39 (29.1%) 26 (66.7%) 13 (33.3%) Radiotherapy 4 (3.0%) 2 (50.0%) 2 (50.0%) Notes: Chi-squared test was employed for categorical variables. * P < 0.05. Abbreviations: FIGO, International Federation of Gynecology and Obstetrics; SCC, squamous cell carcinoma; LVSI, lympho-vascular space invasion; LNM, lymph node metastasis Prognostic significance of PYK2 expression in early-stage cervical patient The survival time was markedly different between the two groups based on the log-rank test. Kaplan-Meier analysis demonstrated that patients in the high-PYK2 group exhibited significantly shorter OS (P = 0.001) and DFS (P = 0.004) than those in the low-PYK2 group (Fig. 2 ). Univariate and multivariate Cox proportional hazard models were performed to identify whether PYK2 protein expression could serve as a prominently independent prognostic factor in cervical cancer (Table 2 ). Several potential prognosis-related factors were tested, including age, FIGO stage, pathological type, tumor size, histologic grade, depth of invasion, lymphovascular space invasion, lymph node metastasis, adjuvant therapy, and PYK2 expression. The result showed that PYK2 expression was indicated as an independent prognostic factor for OS (P = 0.015) and DFS (P = 0.047) eventually. Furthermore, histologic grade and depth of invasion were also significantly associated with OS and DFS. Table 2 Univariate and multivariate of survival for cervical cancer patientsby Cox proportional hazards model Variables Overall Survival Disease-free Survival Univariate analysis a Multivariate analysis b Univariate analysis a Multivariate analysis b HR(95%CI) P-value HR(95%CI) P-value HR(95%CI) P-value HR(95%CI) P-value Age ≤ 52 years 1 0.919 1 0.729 > 52 years 0.950(0.357–2.532) 1.179(0.465–2.986) FIGO stage IB 1 0.205 1 0.273 IIA 2.254(0.641–7.920) 2.001(0.579–6.918) Pathological type SCC 1 0.142 1 0.242 No-SCC 2.337(0.753–7.256) 1.942(0.639–5.903) Tumor Size ≤ 2cm 1 0.998 1 0.785 > 2cm 0.999(0.322–3.098) 1.167(0.384–3.547) Histologic grade Ⅰ-Ⅱ 1 < 0.001* 1 0.001 1 < 0.001* 1 < 0.001* Ⅲ 7.004(2.602–18.853) 5.461(1.939–15.385) 7.018(2.764–17.815) 5.982(2.255–15.874) Depth of invasion < 2/3 1 0.006* 1 0.001* 1 0.017* 1 0.005* ≥ 2/3 4.006(1.490-10.766) 5.430(1.978–14.907) 3.076(1.220–7.754) 3.859(1.515–9.828) LVSI No 1 0.432 1 0.144 Yes 1.423(0.591–3429) 2.158(0.768–6.063) LNM No 1 0.027* 1 0.062 Yes 3.138(1.137–8.662) 2.544(0.953–6.790) Adjuvant therapy No 1 0.506 1 0.392 Chemoradiotherapy 0.374(0.078–1.804) 0.328(0.070–1.543) Chemotherapy 1.274(0.447–3.632) 1.287(0.483–3.430) Radiotherapy - - - - PYK2 expression Low 1 0.004* 1 0.015* 1 0.009* 1 0.047* High 8.643(1.964–38.037) 6.711(1.457–30.907) 4.371(1.438–13.283) 3.237(1.017–10.298) Abbreviations: HR, hazard rate;CI, confidence interval༛FIGO, International Federation of Gynecology and Obstetrics; SCC, squamous cell carcinoma; LVSI, lympho-vascular space invasion; LNM, lymph node metastasis. *P < 0.05. Abbreviations: OS, overall survival; DFS, disease-free survival Abbreviations: si, small interfering; NC, negative control. Prognostic nomogram for 5-year OS and DFS in patients with cervical cancer The nomogram was built on the basis of independent prognostic factors obtained from multivariate analysis of OS and DFS, including PYK2 expression, histologic grade and depth of invasion (Fig. 3 A,B). The calibration plots for the prognostic accuracy were established and certified high consistencies between nomogram-predicted values and the actual observed outcome(Fig. 3 C,D).The Harrell’s concordance index (C-index) were 0.849 and 0.799 for 5-year OS and DFS respectively, which were calculated to validate the result of nomogram. PYK2 downregulation inhibits cervical cancer cell proliferation, invasion and migration ability We performed cellular studies by knocking down PYK2 in Siha and Hela cells, respectively. The downregulation efficiency of PYK2-siRNA was verified using western blotting (Fig. 4 A). As shown in Fig. 4 B, the CCK8 assay demonstrated that PYK2 knockdown significantly decreased cell growth rate compared with cells transfected with negative control plasmid. Additionally, the wound healing assay and transwell assay revealed that co-transfection of Pyk2 siRNA attenuated cell migration and invasion ability in vitro(Fig. 4 C,D). Discussion In this study, we elucidated the clinical significance of PYK2 in early-stage cervical cancer patients for the first time. The high expression level of PYK2 was significantly associated with unfavorable outcomes. We built nomogram models for OS and DFS depend on three prognostic factors, which includes PYK2 expression. Furthermore, by using loss-of-function approaches, we found that PYK2 downregulation hampered the cellular capabilities of proliferation, migration and invasion in an obvious manner. Our study showed that differentiation grade was positively correlated with PYK2 expression, indicating a probability that tumor with higher expression of PYK2 was associated with a greater progressive and invasive risk. PYK2 is found to involved in the biological processes of tumorigenesis, tumor development and invasion via several processes[ 25 , 26 ]. In hepatocellular carcinoma, PYK2 can promote cancer progession by activiting PI3K/AKT signaling pathways[ 19 ]. It was reported that PYK2 can stabilize TAZ protein and negatively regulate the Hippo pathway to promote cell growth and prevent apoptotic cell death in triple-negative breast cancer[ 16 ]. Additionally, overexpressed Pyk2 is uncovered to modulate Wnt/b-catenin pathway by phosphorylating GSK3b Y216 , which initiates and reinforces intestinal tumorigenesis[ 27 ]. PYK2 was also found to take part in metastasis or recurrence in different tumors. As reported in breast cancer, a signalling network between EGFR, c-Met, PYK2 and STAT3 can potentiate EMT and contribute to cancer metastasis[ 7 ]. Moreover, miR-23b may regulate migration by targeting PYK2 via regulation of EMT in hepatocellular carcinoma[ 28 ]. Nevertheless, no significant correlation was found between PYK2 expression and lymph node metastasis in this study with a possibility that the samples included were all relatively early-stage patients with a small probability of lymph node metastasis. As a novel and high-value therapy for cancer, molecule inhibitors of PYK2 obtain continuous attention recent years. PF-00562271 is a widely-concerned PYK2 inhibitor and has been reported to produce robust anti-tumor effects indifferent cancer cells and xenograft tumor models, such as pancreatic cancer, prostate cancer, bone tumors and hepatocellular carcinoma[ 29 – 32 ]. As a second-generation inhibitor of PYK2, VS-6063 was rationally designed to have less potential for drug-drug interaction than PF-00562271[ 33 ]. Both PF-00562271 and VS-6063 were well-tolerated in patients in Phase 1 studies and need further investigation as promising therapeutic drugs[ 34 , 35 ]. We revealed the tumor-promoting role of PYK2 in cervical cancer and further exploration of PYK2 inhibitors are needed to confirm the effect of it on cervical cancer cells. Furthermore,we constructed a prognostic nomogram model to predict the survival rates by integrating PYK2 expression and other clinicopathological prognostic factors. The Harrell’s concordance index indicated a good prognostic ability of the model, which makes it available for clinical use. However, our study had several limitations. Firstly, the clinical data analysis were conducted on a limited cohort size. Secondly, the patients involved were all in the early-stage of cervical cancer with relatively favorable prognosis. Therefore, a larger cohort of patients with high-risk pathological factors are needed in the subsequent study. Thirdly, the underlying molecular mechanisms and signalling pathways of PYK2 involved in the development and progression of cervical cancer remain unknown, which needs further exploration. Conclusion In summary, our study identified that PYK2 was overexpressed in cervical cancer and is associated with its progression. PYK2may represent a prognostic indicator of cervical cancer, which can identify patients at high-risk and serve as a relevant biomarker for predicting the patient outcome. However, further investigation is needed to verify the possibility of applying PYK2 as a potential therapeutic target in clinical practice. Abbreviations PYK2 Proline-rich tyrosine kinase-2; CADTK:Calcium dependent tyrosine kinase; SiRNA:Small interfering RNA; HPV:Human papillomavirus; SCC:Squamous cell carcinoma antigen; FAK:Focal adhesion kinase; TMA:Tissue microarray; IHC:Immunohistochemistry; TMAs:Tissue microarrays; HR:Hazard ratio; CI:Confidence interval; FIGO:International Federation of Gynecology and Obstetrics; C-index:Concordance index Declarations Acknowledgements Not applicable. Author contributions CanZhang, Xinghua Zhu, Aiqin He and Hua Jin were all involved in the study concept and design, drafting of the manuscript, and manuscript revision. Yong Li, Jia Shao, Haibo Xu and Lei Chen contributed to data collection and analysis. Can Zhang and Youli Dan performed experiments. All authors commented on previous versions of the manuscript and approved the final version. Funding This research was supported by the Special Research Found of National Health and Family Planning Commission of Nantong (QA2019027). Availability of data and materials The data sets during and/or analyzed during the current study are available from the corresponding author on reasonable request. Ethics approval and consent to participate All procedures performed in studies involving human participants have obtained the approval of the Ethics Committee of Affiliated Tumor Hospital of Nantong University and have followed the principles of the Declaration of Helsinki. Consent for publication Not applicable. Competing interests The authors declare that they have no competing interests. Author details 1 Department of Gynecology Oncology, Affiliated Tumor Hospital of Nantong University, Nantong, China; 2 Department of Pathology, Affiliated Tumor Hospital of Nantong University, Nantong, China; 3 Medical College of Nantong University, Nantong, China; 4 Cancer Research Centre Nantong, Affiliated Tumor Hospital of Nantong University, Nantong, China. 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Shimizu T, Fukuoka K, Takeda M, Iwasa T, Yoshida T, Horobin J, Keegan M, Vaickus L, Chavan A, Padval M, Nakagawa K. A first-in-Asian phase 1 study to evaluate safety, pharmacokinetics and clinical activity of VS-6063, a focal adhesion kinase (FAK) inhibitor in Japanese patients with advanced solid tumors. Cancer Chemother Pharmacol. 2016;77:997–1003. Infante JR, Camidge DR, Mileshkin LR, Chen EX, Hicks RJ, Rischin D, Fingert H, Pierce KJ, Xu H, Roberts WG, et al. Safety, pharmacokinetic, and pharmacodynamic phase I dose-escalation trial of PF-00562271, an inhibitor of focal adhesion kinase, in advanced solid tumors. J Clin Oncol. 2012;30:1527–33. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-696303","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":39705658,"identity":"e989de53-036a-4d8f-a0e1-b8cb9065deb9","order_by":0,"name":"Can Zhang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA70lEQVRIiWNgGAWjYDACCSjNz97YYPDBwMaOeC2SPYcPFM4oSEsmXovBjbSEzzwfDjE2ENIhP7v52cMvNXZ5BgdyDDfbGBxgZmA/fHQDPi2Mc46ZG8scSy6WPHDG2DjH4A4fA09a2g18WpglEsykJRuYE/sO9pgBtTxjZpDgMcOrhU0i/RtQS31iw2Ee898WBocZGwhp4ZHIMZP82HA4ccIxtgRjBmK0SEjklEkzHDueOLOH+YBhj0FaMhshv8jPSN8m+aOmOrFf/mGDwY8/Nnb87IeP4dUCAsw8KL4jpBwEGH8Qo2oUjIJRMApGLgAAq1xMQYEgMUUAAAAASUVORK5CYII=","orcid":"","institution":"Affiliated Tumor Hospital of Nantong University: Nantong Tumor Hospital","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Can","middleName":"","lastName":"Zhang","suffix":""},{"id":39705659,"identity":"43828f6e-7869-45cc-9665-d254cd7ba228","order_by":1,"name":"Xinghua Zhu","email":"","orcid":"","institution":"Affiliated Tumor Hospital of Nantong University: Nantong Tumor Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xinghua","middleName":"","lastName":"Zhu","suffix":""},{"id":39705660,"identity":"18fea18c-d4c3-4c38-b366-f6668aaaed16","order_by":2,"name":"Yong Li","email":"","orcid":"","institution":"Affiliated Tumor Hospital of Nantong University: Nantong Tumor Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yong","middleName":"","lastName":"Li","suffix":""},{"id":39705661,"identity":"ce441ab3-0902-4260-8d2a-60d193631b67","order_by":3,"name":"Jia Shao","email":"","orcid":"","institution":"Affiliated Tumor Hospital of Nantong University: Nantong Tumor Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jia","middleName":"","lastName":"Shao","suffix":""},{"id":39705662,"identity":"c033dfa5-0938-45ee-9ddf-202d69f0ce61","order_by":4,"name":"Haibo Xu","email":"","orcid":"","institution":"Affiliated Tumor Hospital of Nantong University: Nantong Tumor Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Haibo","middleName":"","lastName":"Xu","suffix":""},{"id":39705663,"identity":"01c90890-5a1c-438a-9355-13997c25af40","order_by":5,"name":"Lei Chen","email":"","orcid":"","institution":"Affiliated Tumor Hospital of Nantong University: Nantong Tumor Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Lei","middleName":"","lastName":"Chen","suffix":""},{"id":39705664,"identity":"8e6c7bfb-da62-4d40-b062-420821b83baa","order_by":6,"name":"Youli Dan","email":"","orcid":"","institution":"Nantong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Youli","middleName":"","lastName":"Dan","suffix":""},{"id":39705665,"identity":"8d7742a7-b034-48ee-885b-2a65b5a52628","order_by":7,"name":"Aiqin He","email":"","orcid":"","institution":"Affiliated Tumor Hospital of Nantong University: Nantong Tumor Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Aiqin","middleName":"","lastName":"He","suffix":""},{"id":39705666,"identity":"e313ae0c-dfcd-4cbd-ae2f-c4fa2fd3044d","order_by":8,"name":"Hua Jin","email":"","orcid":"","institution":"Affiliated Tumor Hospital of Nantong University: Nantong Tumor Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hua","middleName":"","lastName":"Jin","suffix":""}],"badges":[],"createdAt":"2021-07-08 04:55:03","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-696303/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-696303/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":11533415,"identity":"e14f10c8-7d9f-4bc1-90af-6d2febd6cb1f","added_by":"auto","created_at":"2021-07-16 15:11:14","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":495046,"visible":true,"origin":"","legend":"Representative images from immunohistochemistry analysis of PYK2 expression incervical cancer tissues and adjacent noncarcinomaous tissues(ANT). Positive PYK2staining was observed mainly in cervical cancer cellcytoplasm (original magnification ×400). (A) Negative PYK2 staining in ANT. (B) Weak PYK2 staining in cervical cancer tissues. (C) Moderate PYK2 staining in cervical cancer tissues. (D) Strong PYK2 staining in cervical cancer tissues.","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-696303/v1/1c8bb817dfc8cec0a7d7b630.png"},{"id":11533412,"identity":"fc576251-d541-4d10-9727-39bb26d21af3","added_by":"auto","created_at":"2021-07-16 15:11:14","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":18542,"visible":true,"origin":"","legend":"Kaplan-Meier survival curves for overall survival(A) and disease-free survival(B) according to PYK2 expression levels. P-values were determined using the log-rank test.\n\n","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-696303/v1/b8146348636e72aba544b617.png"},{"id":11533414,"identity":"93b63067-eaa0-4f22-b5dd-d4a30fcffe5a","added_by":"auto","created_at":"2021-07-16 15:11:14","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":46929,"visible":true,"origin":"","legend":"Nomogram and calibration plot for OS and DFS prediction in patients with cervical cancer. (A-B) Three independent prognostic factors were integrated into the nomogram to predict 5-year OS survival. (C-D) The calibration plots for predicting OS and DFS at 5 years.\nAbbreviations: OS, overall survival; DFS, disease-free survival\n","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-696303/v1/0fd4f0ef40e5cd3b1d29a59f.png"},{"id":11533942,"identity":"ed62e379-a383-44ff-8763-20e5a1aea354","added_by":"auto","created_at":"2021-07-16 15:14:14","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":80201,"visible":true,"origin":"","legend":"Down-regulation of PYK2 repressed cellular proliferation,motility and invasion. (A) Western blot analysis of PYK2 expression in SiHa and HeLa cells transfected with siRNA. (B) CCK8 assay showed that PYK2 knockdown suppressed proliferation abilities. (C-D) The migration and invasion abilitieswere inhibited in PYK2-knockdown cells, which were evaluated by transwell assays, matrigel invasion assays and wound healing assays. Error bars represent the mean ± SD values ofthree independent experiments. *p \u003c 0.05\nAbbreviations: si, small interfering; NC, negative control.\n","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-696303/v1/55c32d87cb2ca0f6cfc15b8b.png"},{"id":13704597,"identity":"6ce5bfb6-0bad-4c8c-ab1d-ca70320f57e3","added_by":"auto","created_at":"2021-09-17 13:47:07","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3617600,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-696303/v1/aeb03cf3-9a7c-41a5-a094-ef7f7c4b91be.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eHigh Expression of PYK2 is Associated with Poor Prognosis and Cancer Progression in Early- Stage Cervical Carcinoma\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eCervical cancer is the most frequent gynecological cancer and the fourth leading cause of globally cancer-related motility among women, especially in developing countries[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].As known, persistent infection of high-risk oncogenic types of human papillomavirus (HPV) is the primary cause of cervical cancer. However, only a small percentage of patients infected develop into invasive cancer, indicating the carcinogenic effects of other factors[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Although clinical factors, such as stage, squamous cell carcinoma antigen (SCC), lymph node metastasis, may serve as prognostic markers, more clinical trials are needed to confirm and validate the clinical sensitivity[\u003cspan additionalcitationids=\"CR4\" citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Therefore, it is crucial to obtain a better understanding of the underlying molecular mechanism of cervical cancer development and progression, and novel biomarkers are required to investigate, which would provide reliable predictors as well as potential targets for cervical cancer.\u003c/p\u003e \u003cp\u003eProline-rich tyrosine kinase-2 (PYK2) is a non-receptor tyrosine kinase, which belongs to the focal adhesion kinase (FAK) family and regulates cell adhesion, morphology, migration, and invasion via different signal transduction cascades[\u003cspan additionalcitationids=\"CR7 CR8\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. PYK2 consists of three domains associated with tumor progression and can be activated by multiple growth factors, neuropeptides, cytokines, hormones, and chemokines[\u003cspan additionalcitationids=\"CR11 CR12 CR13 CR14\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Overexpression of PYK2 has been reported to promote progression and lead to poor prognosis in many kinds of malignant tumors such as breast cancer, liver cancer, lung cancer and pancreatic cancer[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan additionalcitationids=\"CR17 CR18 CR19 CR20 CR21\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. PYK2 was only found to serve as an onco-suppressor gene in androgen-dependent prostate cancer cells and neuroblastoma cells so far[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. However, the role of PYK2 in cervical cancer is still unclear.\u003c/p\u003e \u003cp\u003eIn our study, we evaluated the role of Pyk2 in early-stage cervical carcinoma and found that the high expression of Pyk2 is associated with poor clinical outcomes in cervical cancer patients. Furthermore, we gained insight into the important functions of this protein in cervical cancer lines.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePatients and clinical samples\u003c/h2\u003e \u003cp\u003eThe study was composed of 268 samples from 134 cervical cancer patients, who treated with radical hysterectomy plus pelvic lymphadenectomy without neoadjuvant chemotherapy or preoperative radiotherapy at Affiliated Tumor Hospital of Nantong University from December 2007 to September 2014. Tissue microarray (TMA) was prepared with the 134 cervical cancer tissues and adjacent normal cervical tissue samples. The study was performed with the approval of the Ethics Committee of Affiliated Tumor Hospital of Nantong University and complied with the Helsinki Declaration.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eCell culture and transfection\u003c/h2\u003e \u003cp\u003e HeLa was obtained from Medical College of Nantong University and SiHa was purchased from the Cell bank of the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). SiHa cells were cultured in minimum essential medium ( HyClone Laboratories, Logan, UT) and Hela cells were cultured in DMEM medium(Biological Industries, Israel), which were all supplemented with 10 % fetal bovine serum (Biological Industries, Israel) and 1 % antibiotics (100 U/mL penicillin and 100 mg/mL streptomycin) in a 5% CO2-humidified atmosphere at 37℃.\u003c/p\u003e \u003cp\u003eTo silence endogenous PYK2 expression, the following Pyk2 small interfering RNA (siRNA) were synthesized by GenePharma (Shanghai, China) with the sequence of 5'-GAUGUGGUCCUGAAUCGUATT-3'. A non-silencing siRNA oligonucleotide that does not recognize any known mammalian gene homolog (GenePharma) was used as a negative control. SiRNA-knockdown were performed with LipoMax\u0026trade; transfection reagent (SUDGEN) following the manufacturer\u0026rsquo;s procedure. The transfection efficiencies were tested by Western blot analysis.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eImmunohistochemistry (IHC) stainingand evaluation\u003c/h2\u003e \u003cp\u003eThe expression of PYK2 was determined using immunohistochemistry (IHC) staining of tissue microarrays (TMAs). A standard immunostaining procedure was performed using Rabbit molyclonal anti-Pyk2 antibody (1:100 dilution; ab32571, Abcam, Cambridge, MA, USA) at 4℃ overnight. Two independent pathologists blind to the clinical or pathologic data assessed the IHC score. The staining intensity was scored according to 4 grades: 0 (No staining), 1 (weak), 2 (moderate), or 3 (intense). The percentage of tumor cells stained was scored as follows: 1, 1\u0026ndash;25%; 2, 26\u0026ndash;50%; 3, 51\u0026ndash;75%; or 4, 76\u0026ndash;100%. The overall IHC score was finally obtained by multiplying the intensity score and percentage scores (ranged from 0 to 12). The samples were considered to be high expression if the score was \u0026gt;\u0026thinsp;4, and defined as low expression if not.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eWestern Blotting\u003c/h2\u003e \u003cp\u003eSamples from cell lysates or tissues lysates were prepared using cold RIPA buffer and separated by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide). Then proteins were transferred onto PVDF (polyvinylidene difluoride) membranes and incubated with primary antibodies including PYK2 (1:2000 dilution; ab32571, Abcam, Cambridge, MA, USA), anti-GAPDH(1:5000 dilution; Santa Cruz, CA, USA).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eCell proliferation assay\u003c/h2\u003e \u003cp\u003eFor CCK8 assay, HeLa and SiHa cells cells were seeded into 96-well plates at density of 3000 cells per well. At the specified time point, 10\u0026micro;l CCK8 solution was added and incubated for 2hours. Cell viability was determined using enzyme-labeled instrument to measure absorbance at 450 nm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eTranswell assay\u003c/h2\u003e \u003cp\u003eA total of 2\u0026times;10\u003csup\u003e4\u003c/sup\u003e HeLa and SiHa cells were seeded into the upper part of a transwell chamber with a pore size of 8 \u0026micro;m (BD, Franklin Lakes, NJ, USA). For the invasion assay, the upper chamber was coated with 60\u0026micro;l prechilled Matrigel. The bottom chambers were filled with 0.5 mL of medium supplemented with 10% FBS and incubated for 24h. The transmembraned cells were fixed with 100% methanol and stained with 0.1% crystal violet for 15min. Finally, the number of migrated and invading cells on the lower side of the membrane was counted under a light microscope.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eWound healing assay\u003c/h2\u003e \u003cp\u003eCells were inculated into 6-well plates and grown in complete medium until 90\u0026ndash;100% confluency. A 5mm-diameter wound was made across the cells using a sterile plastic tip. The complete medium was replaced by serum-free medium then. The migration of the cells was observed under a microscope at 0h, 24h and 48h respectively.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eThe relationship between CPE expression and the clinicopathological characteristics was assessed using the Chi-squared test or Fisher\u0026rsquo;s exact test. Disease-free survival and overall survival were calculated using the Kaplan-Meier method and were compared using the log-rank test. Univariate and multivariate Cox proportional hazards regression models were performed to identify the independent prognostic factors. Results were reported as hazard ratio (HR) and 95% confidence interval (CI). A two-sided P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered to indicate statistical significance. All statistical analysis were obtained by using IBM SPSS software (Statistical Package for the Social Sciences; Version 25.0). Nomograms and calibration plots for OS and DFS based on multivariable analysis were established with R software (version 3.6.1).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003ePYK2 expression with clinicopathological features\u003c/h2\u003e \u003cp\u003ePYK2 was mainly localized in the cytoplasm of cervical cancer cells as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Negative or low PYK2 protein expression was detected in adjacent normal cervical tissue samples. Among the 134 cervical cancer specimens, 62 samples (46.3%) showed high PYK2 protein expression and low expression of PYK2 was observed in 72 tumor samples. The clinicopathologic analysis exhibited that PYK2 expression was strongly correlated with differentiation grade (P\u0026thinsp;=\u0026thinsp;0.027). However, PYK2 expression did not correlate with other clinical features, including age, FIGO (International Federation of Gynecology and Obstetrics) stage, pathological type, tumor size, depth of invasion, lympho-vascular space invasion, lymph node metastasis and adjuvant therapy (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAssociation between PYK2 expression and clinicopathological variables in cervical cancer patients\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVariables\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTotal (n\u0026thinsp;=\u0026thinsp;134)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003ePYK2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eLow expression (%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eHigh expression (%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026le;\u0026thinsp;52 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e64 (47.8%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e31 (48.4%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e33 (51.6%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.240\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;52 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e70 (52.2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e41 (58.6%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e29 (41.4%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFIGO stage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e121 (90.3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e65 (53.7%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e56 (46.3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.993\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIIA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e13 (9.70%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7(53.8%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6 (46.2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePathological type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e115 (85.8%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e61 (53.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e54 (47.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.694\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo-SCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19 (14.2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11 (57.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8 (42.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTumor Size\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026le;\u0026thinsp;2cm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e33 (24.6%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15(45.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e18 (54.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.272\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;2cm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e101 (75.4%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57 (56.4%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e44 (43.6%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHistologic grade\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eI-II\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e110 (80.3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e64 (58.2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e46 (41.8%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.027*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIII\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24 (17.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8 (33.3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e16 (66.7%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDepth of invasion\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;2/3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e99 (73.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e51 (51.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e48 (48.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.387\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026ge;\u0026thinsp;2/3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e35 (26.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e21 (60.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14 (40.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLVSI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e113 (84.3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e60 (53.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e53 (46.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.733\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21(15.7%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12 (57.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9 (42.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLNM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e110 (82.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57 (51.8%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e53(48.2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.342\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24 (17.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15 (62.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9 (37.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAdjuvant therapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e49 (36.6%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23(46.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e26 (53.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.278\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChemoradiotherapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42 (31.3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e21 (50.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21 (50.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChemotherapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e39 (29.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26 (66.7%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13 (33.3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRadiotherapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4 (3.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2 (50.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2 (50.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eNotes: Chi-squared test was employed for categorical variables. * P\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eAbbreviations: FIGO, International Federation of Gynecology and Obstetrics; SCC, squamous cell carcinoma; LVSI, lympho-vascular space invasion; LNM, lymph node metastasis\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003ePrognostic significance of PYK2 expression in early-stage cervical patient\u003c/h2\u003e \u003cp\u003eThe survival time was markedly different between the two groups based on the log-rank test. Kaplan-Meier analysis demonstrated that patients in the high-PYK2 group exhibited significantly shorter OS (P\u0026thinsp;=\u0026thinsp;0.001) and DFS (P\u0026thinsp;=\u0026thinsp;0.004) than those in the low-PYK2 group (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eUnivariate and multivariate Cox proportional hazard models were performed to identify whether PYK2 protein expression could serve as a prominently independent prognostic factor in cervical cancer (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Several potential prognosis-related factors were tested, including age, FIGO stage, pathological type, tumor size, histologic grade, depth of invasion, lymphovascular space invasion, lymph node metastasis, adjuvant therapy, and PYK2 expression. The result showed that PYK2 expression was indicated as an independent prognostic factor for OS (P\u0026thinsp;=\u0026thinsp;0.015) and DFS (P\u0026thinsp;=\u0026thinsp;0.047) eventually. Furthermore, histologic grade and depth of invasion were also significantly associated with OS and DFS.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eUnivariate and multivariate of survival for cervical cancer patientsby Cox proportional hazards model\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eVariables\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eOverall Survival\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c9\" namest=\"c6\"\u003e \u003cp\u003eDisease-free Survival\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eUnivariate analysis\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eMultivariate analysis\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eUnivariate analysis\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003eMultivariate analysis\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHR(95%CI)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHR(95%CI)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHR(95%CI)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eHR(95%CI)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026le;\u0026thinsp;52 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.919\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.729\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;52 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.950(0.357\u0026ndash;2.532)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.179(0.465\u0026ndash;2.986)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFIGO stage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.273\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIIA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.254(0.641\u0026ndash;7.920)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.001(0.579\u0026ndash;6.918)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePathological type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.142\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.242\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo-SCC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.337(0.753\u0026ndash;7.256)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.942(0.639\u0026ndash;5.903)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTumor Size\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026le;\u0026thinsp;2cm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.998\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.785\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;2cm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.999(0.322\u0026ndash;3.098)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.167(0.384\u0026ndash;3.547)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHistologic grade\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eⅠ-Ⅱ\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eⅢ\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.004(2.602\u0026ndash;18.853)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.461(1.939\u0026ndash;15.385)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.018(2.764\u0026ndash;17.815)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e5.982(2.255\u0026ndash;15.874)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDepth of invasion\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;2/3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.006*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.001*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.017*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.005*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026ge;\u0026thinsp;2/3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.006(1.490-10.766)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.430(1.978\u0026ndash;14.907)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.076(1.220\u0026ndash;7.754)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.859(1.515\u0026ndash;9.828)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLVSI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.432\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.144\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.423(0.591\u0026ndash;3429)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.158(0.768\u0026ndash;6.063)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLNM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.027*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.062\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.138(1.137\u0026ndash;8.662)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.544(0.953\u0026ndash;6.790)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAdjuvant therapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.506\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.392\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChemoradiotherapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.374(0.078\u0026ndash;1.804)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.328(0.070\u0026ndash;1.543)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChemotherapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.274(0.447\u0026ndash;3.632)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.287(0.483\u0026ndash;3.430)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRadiotherapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePYK2 expression\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLow\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.004*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.015*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.009*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.047*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHigh\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.643(1.964\u0026ndash;38.037)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.711(1.457\u0026ndash;30.907)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.371(1.438\u0026ndash;13.283)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.237(1.017\u0026ndash;10.298)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003eAbbreviations: HR, hazard rate;CI, confidence interval༛FIGO, International Federation of Gynecology and Obstetrics; SCC, squamous cell carcinoma; LVSI, lympho-vascular space invasion; LNM, lymph node metastasis.\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003e*P\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003eAbbreviations: OS, overall survival; DFS, disease-free survival\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003eAbbreviations: si, small interfering; NC, negative control.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003ePrognostic nomogram for 5-year OS and DFS in patients with cervical cancer\u003c/h2\u003e \u003cp\u003eThe nomogram was built on the basis of independent prognostic factors obtained from multivariate analysis of OS and DFS, including PYK2 expression, histologic grade and depth of invasion (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA,B). The calibration plots for the prognostic accuracy were established and certified high consistencies between nomogram-predicted values and the actual observed outcome(Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC,D).The Harrell\u0026rsquo;s concordance index (C-index) were 0.849 and 0.799 for 5-year OS and DFS respectively, which were calculated to validate the result of nomogram.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003ePYK2 downregulation inhibits cervical cancer cell proliferation, invasion and migration ability\u003c/h2\u003e \u003cp\u003eWe performed cellular studies by knocking down PYK2 in Siha and Hela cells, respectively. The downregulation efficiency of PYK2-siRNA was verified using western blotting (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB, the CCK8 assay demonstrated that PYK2 knockdown significantly decreased cell growth rate compared with cells transfected with negative control plasmid. Additionally, the wound healing assay and transwell assay revealed that co-transfection of Pyk2 siRNA attenuated cell migration and invasion ability in vitro(Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC,D).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn this study, we elucidated the clinical significance of PYK2 in early-stage cervical cancer patients for the first time. The high expression level of PYK2 was significantly associated with unfavorable outcomes. We built nomogram models for OS and DFS depend on three prognostic factors, which includes PYK2 expression. Furthermore, by using loss-of-function approaches, we found that PYK2 downregulation hampered the cellular capabilities of proliferation, migration and invasion in an obvious manner.\u003c/p\u003e \u003cp\u003eOur study showed that differentiation grade was positively correlated with PYK2 expression, indicating a probability that tumor with higher expression of PYK2 was associated with a greater progressive and invasive risk. PYK2 is found to involved in the biological processes of tumorigenesis, tumor development and invasion via several processes[\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. In hepatocellular carcinoma, PYK2 can promote cancer progession by activiting PI3K/AKT signaling pathways[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. It was reported that PYK2 can stabilize TAZ protein and negatively regulate the Hippo pathway to promote cell growth and prevent apoptotic cell death in triple-negative breast cancer[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Additionally, overexpressed Pyk2 is uncovered to modulate Wnt/b-catenin pathway by phosphorylating GSK3b\u003csup\u003eY216\u003c/sup\u003e, which initiates and reinforces intestinal tumorigenesis[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. PYK2 was also found to take part in metastasis or recurrence in different tumors. As reported in breast cancer, a signalling network between EGFR, c-Met, PYK2 and STAT3 can potentiate EMT and contribute to cancer metastasis[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Moreover, miR-23b may regulate migration by targeting PYK2 via regulation of EMT in hepatocellular carcinoma[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Nevertheless, no significant correlation was found between PYK2 expression and lymph node metastasis in this study with a possibility that the samples included were all relatively early-stage patients with a small probability of lymph node metastasis.\u003c/p\u003e \u003cp\u003eAs a novel and high-value therapy for cancer, molecule inhibitors of PYK2 obtain continuous attention recent years. PF-00562271 is a widely-concerned PYK2 inhibitor and has been reported to produce robust anti-tumor effects indifferent cancer cells and xenograft tumor models, such as pancreatic cancer, prostate cancer, bone tumors and hepatocellular carcinoma[\u003cspan additionalcitationids=\"CR30 CR31\" citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. As a second-generation inhibitor of PYK2, VS-6063 was rationally designed to have less potential for drug-drug interaction than PF-00562271[\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Both PF-00562271 and VS-6063 were well-tolerated in patients in Phase 1 studies and need further investigation as promising therapeutic drugs[\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. We revealed the tumor-promoting role of PYK2 in cervical cancer and further exploration of PYK2 inhibitors are needed to confirm the effect of it on cervical cancer cells. Furthermore,we constructed a prognostic nomogram model to predict the survival rates by integrating PYK2 expression and other clinicopathological prognostic factors. The Harrell\u0026rsquo;s concordance index indicated a good prognostic ability of the model, which makes it available for clinical use.\u003c/p\u003e \u003cp\u003eHowever, our study had several limitations. Firstly, the clinical data analysis were conducted on a limited cohort size. Secondly, the patients involved were all in the early-stage of cervical cancer with relatively favorable prognosis. Therefore, a larger cohort of patients with high-risk pathological factors are needed in the subsequent study. Thirdly, the underlying molecular mechanisms and signalling pathways of PYK2 involved in the development and progression of cervical cancer remain unknown, which needs further exploration.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn summary, our study identified that PYK2 was overexpressed in cervical cancer and is associated with its progression. PYK2may represent a prognostic indicator of cervical cancer, which can identify patients at high-risk and serve as a relevant biomarker for predicting the patient outcome. However, further investigation is needed to verify the possibility of applying PYK2 as a potential therapeutic target in clinical practice.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003ePYK2\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eProline-rich tyrosine kinase-2; CADTK:Calcium dependent tyrosine kinase; SiRNA:Small interfering RNA; HPV:Human papillomavirus; SCC:Squamous cell carcinoma antigen; FAK:Focal adhesion kinase; TMA:Tissue microarray; IHC:Immunohistochemistry; TMAs:Tissue microarrays; HR:Hazard ratio; CI:Confidence interval; FIGO:International Federation of Gynecology and Obstetrics; C-index:Concordance index\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCanZhang, Xinghua Zhu, Aiqin He and Hua Jin were all involved in the study concept and design, drafting of the manuscript, and manuscript revision. Yong Li, Jia Shao, Haibo Xu and Lei Chen contributed to data collection and analysis. Can Zhang and Youli Dan performed experiments. All authors commented on previous versions of the manuscript and approved the final version.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by the Special Research Found of National Health and Family Planning Commission of Nantong (QA2019027).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data sets during and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll procedures performed in studies involving human participants have obtained the approval of the Ethics Committee of Affiliated Tumor Hospital of Nantong University and have followed the principles of the Declaration of Helsinki.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor details\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003e Department of Gynecology Oncology, Affiliated Tumor Hospital of Nantong University, Nantong, China; \u003csup\u003e2\u003c/sup\u003eDepartment of Pathology, Affiliated Tumor Hospital of Nantong University, Nantong, China;\u003csup\u003e3\u003c/sup\u003eMedical College of Nantong University, Nantong, China; \u003csup\u003e4\u003c/sup\u003eCancer Research Centre Nantong, Affiliated Tumor Hospital of Nantong University, Nantong, China.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eTorre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. 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Br J Cancer. 2013;109:1252\u0026ndash;63.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMeans AL. PYK2 at the Intersection of Signaling Pathways in Pancreatic Cancer. Cell Mol Gastroenterol Hepatol. 2019;8:651\u0026ndash;2.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShen T, Guo Q. Role of Pyk2 in Human Cancers. Med Sci Monit. 2018;24:8172\u0026ndash;82.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMaudsley S, Davidson L, Pawson AJ, Freestone SH, L\u0026oacute;pez de Maturana R, Thomson AA, Millar RP. Gonadotropin-releasing hormone functionally antagonizes testosterone activation of the human androgen receptor in prostate cells through focal adhesion complexes involving Hic-5. Neuroendocrinology. 2006;84:285\u0026ndash;300.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBosch R, Dieguez-Gonzalez R, Moreno MJ, Gallardo A, Novelli S, Espinosa I, C\u0026eacute;spedes MV, Pav\u0026oacute;n M, Briones J, Gra\u0026ntilde;ena A, et al. Focal adhesion protein expression in human diffuse large B-cell lymphoma. Histopathology. 2014;65:119\u0026ndash;31.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNaser R, Aldehaiman A, D\u0026iacute;az-Galicia E, Arold ST. \u003cb\u003eEndogenous Control Mechanisms of FAK and PYK2 and Their Relevance to Cancer Development\u003c/b\u003e. \u003cem\u003eCancers (Basel)\u003c/em\u003e 2018, 10.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhu X, Bao Y, Guo Y, Yang W. \u003cb\u003eProline-Rich Protein Tyrosine Kinase 2 in Inflammation and Cancer\u003c/b\u003e. \u003cem\u003eCancers (Basel)\u003c/em\u003e 2018, 10.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGao C, Chen G, Kuan SF, Zhang DH, Schlaepfer DD, Hu J. \u003cb\u003eFAK/PYK2 promotes the Wnt/β-catenin pathway and intestinal tumorigenesis by phosphorylating GSK3β\u003c/b\u003e. \u003cem\u003eElife\u003c/em\u003e 2015, 4.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCao J, Liu J, Long J, Fu J, Huang L, Li J, Liu C, Zhang X, Yan Y. microRNA-23b suppresses epithelial-mesenchymal transition (EMT) and metastasis in hepatocellular carcinoma via targeting Pyk2. 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Cancer Sci. 2017;108:1347\u0026ndash;56.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBagi CM, Christensen J, Cohen DP, Roberts WG, Wilkie D, Swanson T, Tuthill T, Andresen CJ. Sunitinib and PF-562,271 (FAK/Pyk2 inhibitor) effectively block growth and recovery of human hepatocellular carcinoma in a rat xenograft model. Cancer Biol Ther. 2009;8:856\u0026ndash;65.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJones SF, Siu LL, Bendell JC, Cleary JM, Razak AR, Infante JR, Pandya SS, Bedard PL, Pierce KJ, Houk B, et al. A phase I study of VS-6063, a second-generation focal adhesion kinase inhibitor, in patients with advanced solid tumors. Invest New Drugs. 2015;33:1100\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShimizu T, Fukuoka K, Takeda M, Iwasa T, Yoshida T, Horobin J, Keegan M, Vaickus L, Chavan A, Padval M, Nakagawa K. A first-in-Asian phase 1 study to evaluate safety, pharmacokinetics and clinical activity of VS-6063, a focal adhesion kinase (FAK) inhibitor in Japanese patients with advanced solid tumors. Cancer Chemother Pharmacol. 2016;77:997\u0026ndash;1003.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eInfante JR, Camidge DR, Mileshkin LR, Chen EX, Hicks RJ, Rischin D, Fingert H, Pierce KJ, Xu H, Roberts WG, et al. Safety, pharmacokinetic, and pharmacodynamic phase I dose-escalation trial of PF-00562271, an inhibitor of focal adhesion kinase, in advanced solid tumors. J Clin Oncol. 2012;30:1527\u0026ndash;33.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"PYK2, early-stage cervical carcinoma, prognosis, nomogram, biomaker","lastPublishedDoi":"10.21203/rs.3.rs-696303/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-696303/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e Proline-rich tyrosine kinase-2 (PYK2), also known as calcium dependent tyrosine kinase (CADTK), regulates different signal transduction cascades that control cell proliferation, migration and invasion. However, the role of PYK2 in cervical cancer remains to be elucidated.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e PYK2 expression was detected in tissue microarray and cervical cancer cell lines. Statistical analysis was performed to evaluate its clinicopathological significance. Small interfering RNA (SiRNA) was employed to suppress endogenous PYK2 expression in cervical cancer cells to observe the biological function.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003ePYK2 expression was up-regulated in cervical cancer specimens compared with paired adjacent normal cervical tissue samples. Statistical analyses indicated that PYK2 expression might be an independent prognostic indicator for patients with early-stage cervical cancer. A nomogram model was constructed based on PYK2 expression and other clinicopathological risk factors, and it performed well in predicting patients survival. In cellular studies, down-regulation of PYK2 remarkably inhibited cellular proliferation, migration and invasion.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e PYK2 expression possessed the potential to serve as a novel prognostic marker\u003c/p\u003e\u003cp\u003ein cervical cancer patients.\u003c/p\u003e","manuscriptTitle":"High Expression of PYK2 is Associated with Poor Prognosis and Cancer Progression in Early- Stage Cervical Carcinoma","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-07-16 15:11:12","doi":"10.21203/rs.3.rs-696303/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"45cf419b-4f1f-478f-ad3d-e822381e8921","owner":[],"postedDate":"July 16th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":5772523,"name":"Surgery"},{"id":5772524,"name":"Oncology"}],"tags":[],"updatedAt":"2021-08-23T17:04:22+00:00","versionOfRecord":[],"versionCreatedAt":"2021-07-16 15:11:12","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-696303","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-696303","identity":"rs-696303","version":["v1"]},"buildId":"GqpaHPwrfC8PjnIFayRh5","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}