The expression and clinical significance of circ_0007167 in patients with gastric cancer

The expression and clinical significance of circ_0007167 in patients with gastric cancer | 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 Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The expression and clinical significance of circ_0007167 in patients with gastric cancer Feilong Zhu, Yingting Wu, Zhen Ye, Xiaotian Wu, Xueping Cao, Huifen Chen This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4303861/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 Gastric cancer (GC) is one of the commonest malignant tumors worldwide including China. Data from epidemiological surveys showed that the new cases of GC were globally increased by 1 million per year, and 800,000 people died from gastric cancer every year. In recent years, it has been found that cyclic RNA plays an important role in the occurrence of GC, and it may be used as an evaluation index for the diagnosis and prognosis of GC. This study was conducted to assess the clinical application value of circ_0007167 for diagnosis and prognosis of GC. Methods GC group was collected from September 1, 2019 to November 30, 2020 at the Affiliated Hospital of Nantong University. There were 108 patients with GC confirmed by imaging and pathology, including 20 pairs of plasma from GC patients before and one week after surgery. There were 40 patients with benign gastric lesions and 79 healthy individuals from the physical examination center of the Affiliated Hospital of Nantong University. The expression levels of circ_0007167 in the plasma samples of the three groups were detected using qRT-PCR following validation of the method. The correlations between changes in plasma circ_0007167 levels and the clinical pathological parameters of GC patients were statistically analyzed. Meanwhile, the ROC curves of plasma circ_0007167, CEA and CA19-9 levels were analyzed to study the clinical value of single and combined detection in the diagnosis and prognosis of GC. Results The expression level of circ_0007167 in the plasma of patients with GC was significantly higher than that in the benign gastric lesions group and the healthy control group (both p < 0.001). There was no significant difference between benign gastric lesion group and healthy control group (p = 0.328). Further clinicopathologic features showed that plasma circ_0007167 was not related to gender (p = 0.687), age (p = 0.841), and differentiation degree of tumor (p = 0.9), but related to tumor size (p < 0.05), T stage (p < 0.001), metastasis (p < 0.001) and nerve/vascular invasion (p = 0.006). The diagnostic efficacy of ROC curve analysis showed that the area under the curve (AUC) of circ_0007167 was 0.881, significantly higher than that of CA19-9 (AUC = 0.679) and CEA (AUC = 0.698), and the diagnostic efficacy of circ_0007167 was better than that of traditional tumor markers CEA and CA19-9. There was no significant difference in diagnostic efficacy between CEA and CA19-9, but when circ_0007167 was combined with CEA, AUC was 0. 893; when circ_0007167 was combined with CA19-9, AUC was 0. 891; when circ_0007167 was combined with CEA and CA19-9 as diagnostic markers, diagnostic efficiency is the best with AUC = 0.896. The expression of circ_0007167 in plasma of GC patients decreased significantly after operation compared to that before operation (p < 0.001). Conclusion Plasma circ_0007167 in patients with GC is superior to traditional tumor markers CEA and CA19-9. Combined detection of circ_0007167 with CEA and CA19-9 can improve the assisted diagnosis of early GC. Moreover, the plasma circ_0007167 was decreased significantly following surgery, suggesting that plasma circ_0007167 may serve as a new biomarker for the diagnosis, treatment, and prognosis evaluation of GC. circRNA circ_0007167 gastric cancer diagnosis Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1 Introduction Gastric cancer (GC) is a malignant tumor that originates from the epithelium of the stomach mucosa, ranking fifth in global cancer incidence rates [ 1 ], and most of its occurrences are found in East Asia [ 2 ]. The main risk factors for GC are Helicobacter pylori infection, while other risk factors include geographical location, age, gender, smoking, salted meat consumption and genetics [ 3 ]. GC is often asymptomatic in the early stages, and about 80% of patients have no obvious symptoms at the early stage of the disease, leading to easy missed diagnosis at the early stage [ 4 ]. Due to the lack of reliable and effective early diagnosis techniques, many patients are still diagnosed at an advanced stage, resulting in a 5-year survival rate of only 30% [ 5 ]. Circular RNA (circRNA) was first discovered in the 1970s [ 6 ]. More than 40 years ago, people first observed that eukaryotic RNA can exist in a circular form using electron microscopy [ 7 ]. CircRNA is a covalently closed circular structure formed by the direct connection of the 5' and 3' ends, without a 3' tail or a 5' cap structure [ 8 ]. In recent years, more and more studies have found that circRNA is closely related to the development of GC. Liu found that the expression level of circYAP1 in GC tissues was significantly lower than that in adjacent tissues, and the survival time of patients with low expression of circYAP1 was shorter than that of patients with high expression of circYAP1. CircYAP1 exerts tumor suppressor effects in GC cells by targeting the miR-367-5p/p27 axis [ 9 ]. Jie found that the expression of circMRPS35 in GC tissues was significantly down-regulated, which was associated with tumor size, TNM stage, and lymphatic metastasis, and was positively correlated with the expression of FOXO1/3a [ 10 ]. Over the past few decades, the global incidence and mortality rates of GC have been decreasing, which is closely related to the decline in Helicobacter pylori infection rates, as well as improvements in hygiene conditions and the widespread use of antibiotics [ 11 ]. Although the incidence is decreasing, the burden of GC patients worldwide will increase in the coming years due to the continuous growth and aging of the world's population [ 12 ]. The 5-year survival rate is usually higher in patients with early GC (> 60%), while patients with GC with local and distant metastases have a significantly lower 5-year survival rate. Due to the atypical and occult nature of early GC, more than 60% of patients are already in the advanced stage at diagnosis [ 13 ]. However, commonly used biomarkers for early GC screening, such as carcinoembryonic antigen (CEA) and carbohydrate antigen 19 − 9 (CA19-9), have low sensitivity and specificity rates [ 14 ]. Currently, endoscopic biopsy combined with pathological examination remains the gold standard for GC diagnosis, but its invasive nature and high cost limit its clinical acceptance [ 15 ]. Therefore, the low rate of early diagnosis is the main reason for poor prognosis in GC, so there is an urgent need to find non-invasive detection methods and biomarkers with high sensitivity and specificity to improve early diagnosis and survival rates. Based on the above theoretical foundation, this study aims to explore the expression and clinical significance of circ_0007167 in GC. 2 Materials and methods 2.1 Subjects One hundred and eight plasma samples were collected from GC patients in the affiliated hospital of Nantong University. All patients had not received radiotherapy, chemotherapy or other treatment before hospitalization. Forty patients with benign gastric lesions and 79 healthy subjects were also recruited in the affiliated hospital of Nantong University. All plasma samples were the leftover samples from routine medical tests in the Department of Medical Examination. All subjects in this study were reviewed and approved by the Medical Ethics Committee of the affiliated hospital of Nantong University, and the subjects provided informed consent. 2.2circRNA Sequencing Through tissue-based circRNA sequencing, we identified 815 circRNAs with a fold change greater than 2 and p<0.05. By comparing the circRNA sequencing results of three additional pairs of tissues in the GEO database, we intersected them to obtain 73 circRNAs molecules that were consistently up-regulated across different sequencing datasets. By analyzing the correlation between module feature genes and clinical traits, we identified functionally distinct modules. We selected the module positively associated with GC carcinogenesis and intersected it with the two sequencing datasets, leading us to the identification of circRNA_0007167 as a candidate involved in GC development and exhibiting significant up-regulation for further investigation. 2.3 Realtime PCR and determination of CA19-9, CEA levels Total RNA was extracted from cells using TRIzol reagent (Thermo Fisher Scientifc, California, USA). A PrimeScript RT Reagent Kit (Thermo Fisher Scientifc, California, USA) was used for RNA generation. Quantitative real-time PCR (qRT-PCR) was performed by a SYBR Premix Ex Taq II (TaKaRa, Japan) via a 7900HT real-time PCR system (Thermo Fisher Scientifc, California, USA). The PCR reaction uses 18S rRNA as an internal reference and employs the 2-ΔΔct quantitative method. Primers are listed as below: circ_0007167-F: 5'-TACCGTCAAATCCCAAGCCC-3' circ_0007167-R: 5'-AGACATGAGCAGACCAGAATCT-3' 18s-F: 5'-CGGCTACCACATCCAAGGAA-3' 18s-R: 5'-GCTGGAATTACCGCGGCT-3' Detection of CA19-9 and CEA levels using Abbott i2000 chemiluminescence analyzer (Abbott /USA) in strict accordance with the instructions for use. 2.4 Statistical analyses Statistical analyses were performed by the SPSS 21.0 (Armonk, USA) and GraphPad Prism 8.0 (San Diegl, USA) software. The relative expression of circRNA_0007167 in plasma and the contents of CEA and CA19-9 in different groups were expressed by the median (interquartile range) [M (P25, P75)]. The differences in the expression levels of the three groups were compared by one-way analysis of variance, and pairwise comparisons were performed using the LSD method. The categorical variables were expressed by number (percentage) and compared between groups using the χ2 test. The diagnostic value of individual and combined diagnostic indicators in clinical diagnosis of gastric cancer was evaluated using the receiver operating characteristic (ROC) curve and the 95% confidence interval (95%CI) of the area under the curve (AUC). The comparison of the AUC areas was performed using paired Z-tests. The comparison of diagnostic evaluation indicators (accuracy, specificity, sensitivity, positive predictive value, and negative predictive value) between groups was performed using chi-square tests and Boffernoni multiple comparisons. P values less than 0.05 were considered statistically significant (*, p<0.05;**, p<0.01༛***, p<0.001). 3 Results 3.1 Identification of Deregulated circRNA in GC Tissues To investigate the expression profiles of circRNAs in GC tissues, we conducted high-throughput sequencing in three GC tissues vs. three matched noncancerous tissues and identified a total of 25,303 circRNA targets, including 20,036 known circRNAs and 5,267 undefined circRNAs. The heatmap was depicted as a direct approach to visualize the distributions of the dataset for circRNA profiles (Fig. 1A). Volcano plots depicted 2,007 differentially expressed circRNAs in the GC tissues (Flod Change>2.5, p<0.05) (Fig. 1B). To identify the target circRNA, we searched the circRNA database for their chromosomal locations, transcript information, ORF, IRES, and other information. We also read the literature and selected five circRNAs for PCR verification. In both tissues and plasma, circ_0007167 showed significant differences and high correlation, so we ultimately selected circ_0007167 as the target molecule. (A) ( B) Figure 1 Identification of deregulated circular RNAs (circRNAs) in gastric cancer (GC) tissues. ( A ) Clustered heatmap. Each row represents a circRNA, and each column represents a tissue sample. The color scale reflects the log2 signal strength from green (low intensity) to black (medium intensity) to red (strong intensity). ( B ) Volcano plots. The red points in plot indicate the differentially upregulated expression of circRNAs with statistical significance while the green points indicate the downregulated circRNAs. 3.2 Detection of relative expression level of plasma circ_0007167 108 plasma samples were collected from gastric cancer patients, including 70 males and 38 females, aged from 36 to 85 years old. 40 plasma samples were collected from patients with benign gastric lesions, including 25 males and 15 females, aged from 43 to 85 years old. 79 plasma samples were collected from healthy subjects, including 42 males and 37 females, aged from 40 to 89 years old. Intergroup comparison showed that the genders were matched (p > 0.05) (Table 1 ). Table 1 Comparison of the gender composition of the study subjects Gender GC( n = 108) Gastric benign lesions( n = 40) Healthy controls( n = 79) male 70(64.81) 25(62.50) 42(53.16) female 38(35.19) 15(37.50) 37(46.84) χ 2 2.682 p-Value 0.262 The relative expression levels of plasma circ_0007167 were measured by qRT-PCR in 108 gastric cancer patients, 40 patients with benign gastric lesions, and 79 healthy subjects. The data were processed using SPSS 21.0. The relative expression levels of plasma circ_0007167 in the gastric cancer group, benign gastric lesion group, and healthy control group were 5.174 (2.741, 7.898), 1.641 (1.341, 2.086), and 1.027 (0.724, 1.561), respectively. The results of multiple comparisons using analysis of variance showed that the relative expression levels of plasma circ_0007167 were significantly higher in the gastric cancer group than in the benign gastric lesion group and healthy control group (p < 0.001). There was no significant difference in the relative expression levels of plasma circ_0007167 between the benign gastric lesion group and healthy control group (p = 0.328) (Fig. 2 ). 3.3 Correlation analysis of circ_0007167 expression and the clinicopathological parameters in GC patients As shown in Table 2 , the expression of circ_0007167 in GC tissues was significantly correlated with tumor size (p < 0.05), T staging (p < 0.001), lymph node metastasis (p < 0.001), and nerve/vascular invasion (p = 0.006). However, we did not find any association between the circ_0007167 expression and other clinicopathological parameters, such as gender (p = 0.687), age (p = 0.841), and differentiation degree (p = 0.699). Table 2 The association between circ_0007167 expression and the clinicopathological parameters in GC patients. Characteristics n High expression (n = 108) Low expression (n = 54) χ 2 P-Value Gender male 70 34(48.57) 36(51.43) 0.162 0.687 female 38 20(52.63) 18(47.37) Age(years) <60 39 20(51.28) 19(48.72) 0.040 0.841 ≥ 60 69 34(49.28) 35(50.72) Tumor size(cm) <5 68 29(42.65) 39(57.35) 3.971 0.046* ≥ 5 40 25(62.50) 15(37.50) Degree of differentiation High-moderate 60 31(51.67) 29(48.33) 0.150 0.699 Low 48 23(47.92) 25(52.08) Tumor depth T1-T2 56 42(75.00) 14(25.00) 29.077 <0.001*** T3-T4 52 12(23.08) 40(76.92) Lymph node metastasis Yes 48 38(79.17) 10(20.93) 29.400 <0.001*** No 60 16(26.67) 44(73.33) Nerve/vascular invasion Yes 42 28(66.67) 14(33.33) 7.636 0.006 No 66 26(39.39) 40(60.61) 3.4Evaluation of diagnostic performance of plasma circ_0007167, CA19-9, and CEA Based on the expression levels of plasma circ_0007167, CA19-9, and CEA in 108 GC patients and 79 healthy individuals, ROC curves were plotted. It was found that the diagnostic performance of circ_0007167 was the best, with an area under the ROC curve of 0.881 (95% CI: 0.829–0.943; p < 0.001). When CEA was used as a diagnostic indicator, the area under the ROC curve was 0.698 (95% CI: 0.620–0.775; p < 0.001). When CA19-9 was used as a diagnostic indicator, the area under the ROC curve was 0.679 (95% CI: 0.601–0.756; p < 0.001) (Fig. 3 ). Furthermore, a pairwise comparison of the areas under the ROC curves showed that the diagnostic performance of circ_0007167 was significantly better than that of CEA or CA19-9 (p < 0.001 for both comparisons). There was no significant difference in the diagnostic performance between CEA and CA19-9(p = 0.463)(Table 3 ). Table 3 Comparison of areas under the ROC curves for diagnosis using three indicators Comparison group AUC difference Z P-Value circ_0007167 CEA 0.184 4.209 <0.001 circ_0007167 CA19-9 0.203 4.756 <0.001 CEA CA19-9 0.019 0.463 0.463 3.5 Evaluation of the diagnostic performance of combined detection of plasma circ_0007167, CA19-9, and CEA After combining circ_0007167 with CA19-9 and CEA for diagnosis and plotting ROC curves, it was found that when circ_0007167 was combined with CEA as a diagnostic indicator, the area under the ROC curve was 0.893 (95% CI: 0.843–0.943; p < 0.001); when circ_0007167 was combined with CA19-9 as a diagnostic indicator, the area under the ROC curve was 0.891 (95% CI: 0.842–0.940; p < 0.001); when circ_0007167, CEA, and CA19-9 were combined as diagnostic indicators, the area under the ROC curve was 0.896 (95% CI: 0.846–0.945; p 0.05), indicating that the diagnostic efficacies of circ_0007167 combined with CEA or CA19-9, as well as all three combined, are equivalent (Table 4 ). Table 4 Comparison of areas under the ROC curves for three combined diagnostic methods Comparison group AUC difference Z P-Value circ_0007167 + CEA circ_0007167 + CA19-9 0.002 0.193 0.847 circ_0007167 + CEA circ_000716 + CEA + CA19-9 -0.003 -0.366 0.714 circ_0007167 + CA19-9 circ_000716 + CEA + CA19-9 -0.004 -0.439 0.661 Further analysis of the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of plasma circ_0007167, CEA, and CA19-9 as individual diagnostic indicators and combined indicators for GC patients showed that the sensitivity of circ_0007167 was 86.11%, which was superior to CEA (65.74%) and CA19-9 (50.93%). The sensitivity was improved by pairwise and triplet combinations, with the highest sensitivity of 92.59% achieved by the triplet combination. Table 5 Evaluation of the diagnostic values of combination of circ_0007167, CEA and CA19-9. SEN(%) SPE(%) ACCU(%) PPV(%) NPV(%) circ_0007167 86.11 (93/108) 87.34 (69/79) 86.63 (162/187) 90.29 (93/103) 82.14 (69/84) CEA 65.74 (71/108) 65.82 (52/79) 65.78 (123/187) 72.45 (71/98) 58.43 (52/89) CA19-9 50.93 (55/108) 74.68 (59/79) 60.96 (114/187) 73.33 (55/75) 52.68 (59/112) circ_0007167 + CEA 91.67 (99/108) 49.37 (39/79) 73.80 (138/187) 71.22 (99/139) 81.25 (39/48) circ_0007167 + CA19-9 87.04 (94/108) 55.70 (44/79) 73.80 (138/187) 72.87 (94/129) 75.86 (44/58) circ_0007167 + CEA + CA19-9 92.59 (100/108) 37.97 (30/79) 69.52 (130/187) 67.11 (100/149) 78.95 (30/38) SEN, sensitivity; SPE, specificity; ACCU, overall accuracy; PPV, positive predictive value; NPV, negative predictive value 3.6 Monitoring ability of plasma circ_0007167 for GC disease progression To dynamically detect the expression level of circ_0007167 in the plasma of 20 patients newly diagnosed with GC, and to analyze the differences between the two groups using paired t-test analysis. The results showed that the plasma circ_0007167 level decreased overall in GC patients after surgery and was statistically significant (p < 0.001) (Fig. 5 ). 4 Discussion Gastric cancer (GC) is one of the most common cancers worldwide, with over 1 million new cases reported in 2018. Nearly two-thirds of these cases occur in developing countries, especially in East Asia, where the incidence rate is the highest. The risk of GC is 2–3 times higher in men than in women, and the risk increases with age, with the highest incidence rate occurring between the ages of 60 and 70 years old [ 16 ]. Due to the asymptomatic nature of early GC, patients often present with symptoms such as weight loss and abnormal pain by the time they are diagnosed with advanced GC, leading to a poor overall survival rate (OS) of less than 30% [ 17 ]. However, in Japan and Korea, large-scale screening has increased the early diagnosis rate of GC, resulting in an OS of over 60% [ 18 ]. Therefore, early detection, diagnosis, and treatment of GC may be the most effective way to address this issue. It is crucial to identify new potential diagnostic biomarkers and therapeutic targets to improve the diagnosis and treatment of GC [ 19 ]. This study aims to identify circular RNA markers associated with GC that may contribute to its diagnosis and treatment. In recent years, circRNA has become a research hotspot in the diagnosis and treatment of GC. More and more high-throughput sequencing and microarray gene chip technologies have shown significant differences in the expression levels of circRNA between GC patients and healthy individuals [ 20 – 21 ]. Additionally, the study of circRNA in peripheral blood has the advantages of convenient collection, minimal trauma, and high patient acceptance, making it a potential new biomarker for tumors. In view of the fact that there are few studies on GC diagnosis using serum or plasma circ_0007167 as biomarkers, this research group has selected circ_0007167 as the research object through gene chip screening in the early stage to explore its significance in GC auxiliary diagnosis. After statistical processing of the relative expression quantity of plasma circ_0007167 by 2 −△△CT method, it was found that the content of plasma circ_0007167 in GC patients was significantly higher than that in patients with benign gastric lesions and healthy controls, while there was no significant difference between the benign gastric lesion group and the healthy control group. Further analysis of clinical pathological characteristics found that the level of plasma circ_0007167 was not related to patient's gender, age, and differentiation degree, but was related to tumor size, T staging, lymph node metastasis and nerve/vascular invasion. It shows that the level of plasma circ_0007167 is associated with the progression of GC patients to a certain extent. ROC curve analysis of diagnostic efficacy found that the area under the curve (AUC) of circ_0007167 was 0.881, significantly higher than that of CA19-9 and CEA. The diagnostic efficacy of circ_0007167 was better than that of traditional markers CEA and CA19-9. An AUC of 0.7–0.9 suggests good clinical diagnostic value, while an AUC of 0.5–0.7 indicates poor clinical diagnostic value [ 22 ]. Further pairwise comparison of the AUC showed that the diagnostic efficacy of circ_0007167 was higher than that of CEA or CA19-9, while there was no significant difference in the diagnostic efficacy between CEA and CA19-9. When circ_0007167 was combined with CA19-9 and CEA for diagnosis, the ROC curve showed that the AUC was 0.893 when circ_0007167 was combined with CEA, 0.891 when circ_0007167 was combined with CA19-9, and 0.896 when circ_0007167, CEA, and CA19-9 were combined as diagnostic indicators, with the best diagnostic efficacy. The sensitivity of circ_0007167 combined with CEA was 91.67%, and the specificity was 49.37%. The sensitivity of circ_0007167 combined with CA19-9 was 87.04%, and the specificity was 55.70%. The sensitivity of all three markers combined was 92.59%, and the specificity was 37.97%, with a decrease in specificity. Therefore, the combination of circ_0007167, CA19-9, and CEA can improve the diagnostic efficacy. Surgical resection is the main treatment for GC, but how to improve postoperative survival rates remains a hot topic [ 23 ]. We found through simultaneous follow-up of GC patients that preoperative plasma circ_0007167 expression was higher, but it decreased postoperatively, suggesting that plasma circ_0007167 may have the ability to monitor GC progression. In summary, the high expression of plasma circ_0007167 in GC patients can be used as a new biomarker for GC diagnosis and prognostic monitoring. In the later stage, we will further expand the sample size and conduct follow-up studies to analyze the correlation between survival time and plasma circ_0007167 expression levels in GC patients, further validating the clinical application value of plasma circ_0007167 in GC patients. Peripheral blood tumor markers are important indicators reflecting tumor progression and prognosis in clinical practice [ 24 ]. In the future, we will further study the biological function of circ_0007167 in GC, providing new ideas for the diagnosis and treatment of GC. Declarations Authors’ Contributions: All authors contributed to the conception and the main idea of the work. FLZ wrote the manuscript and performed the experiences. FLZ, YTW, ZY, XTW, XPC and HFC analyzed the data and edited the manuscript. All authors reviewed the results and approved the final version of the manuscript. Ethics Approval and Informed Consent Statement: The study was approved by the ethics committee of the Affiliated Hospital of Nantong University. Availability of Data and Materials: The datasets generated for this study can be found in GEO database, GSE131414. Funding Statement : This project was supported by grants from the National Natural Science Foundation of China (81871720，82072363). Conflicts of Interest: The authors declare that they have no conflicts of interest to report regarding the present study. References Fock KM. Review article: the epidemiology and prevention of gastric cancer. J Aliment Pharmacol Ther. 2014;40(3):250–60. 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J Biosci Rep. 2019; 39(6). Tang T, Fu W, Sun K, Rong H, Wang D, Cao H. CircRNA microarray profiling identifies a novel circulating biomarker for detection of gastric cancer. J Mol Cancer. 2018;17(1):137. Toft JH, Økland I, Dalen I. ROC-curves-fundamentals for proper use. J Endocr. 2022;76(2):505. Ajani JA, Lee J, Sano T, Janjigian YY, Fan D, Song S. Gastric adenocarcinoma. J Nat Rev Dis Primers. 2017;3(2):17036. So JBY, Kapoor R, Zhu F, Koh C, Zhou L, Zou R, Tang YC, Goo PCK, Rha SY, Chung HC, et al. Development and validation of a serum microRNA biomarker panel for detecting gastric cancer in a high-risk population. J Gut. 2021;70(5):829–37. Additional Declarations No competing interests reported. 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 Advisory Board 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-4303861","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":300687540,"identity":"345eb227-7b00-4c7f-9512-086f89e18dba","order_by":0,"name":"Feilong Zhu","email":"","orcid":"","institution":"Shanghai First Maternity and Infant Hospital","correspondingAuthor":false,"prefix":"","firstName":"Feilong","middleName":"","lastName":"Zhu","suffix":""},{"id":300687545,"identity":"6aaf8bcb-4f02-4563-9032-e3c1f735c0e4","order_by":1,"name":"Yingting Wu","email":"","orcid":"","institution":"Shanghai First Maternity and Infant Hospital","correspondingAuthor":false,"prefix":"","firstName":"Yingting","middleName":"","lastName":"Wu","suffix":""},{"id":300687551,"identity":"41ee30f1-d5ae-4736-80ba-468d5ee6ef4e","order_by":2,"name":"Zhen Ye","email":"","orcid":"","institution":"Shanghai First Maternity and Infant Hospital","correspondingAuthor":false,"prefix":"","firstName":"Zhen","middleName":"","lastName":"Ye","suffix":""},{"id":300687557,"identity":"60e6efce-ad4a-44a0-afd3-682b69bbcc7f","order_by":3,"name":"Xiaotian Wu","email":"","orcid":"","institution":"Shanghai First Maternity and Infant Hospital","correspondingAuthor":false,"prefix":"","firstName":"Xiaotian","middleName":"","lastName":"Wu","suffix":""},{"id":300687563,"identity":"341f62da-fc23-43f9-bc4c-ceb326001952","order_by":4,"name":"Xueping Cao","email":"","orcid":"","institution":"Shanghai First Maternity and Infant Hospital","correspondingAuthor":false,"prefix":"","firstName":"Xueping","middleName":"","lastName":"Cao","suffix":""},{"id":300687567,"identity":"459474fe-5e38-4796-9a18-aac5c3ce047b","order_by":5,"name":"Huifen Chen","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAyUlEQVRIiWNgGAWjYBACfmbmg4///rGRs29vIFKLZDtbsgFvQ5qxAc8BIrUYnOcxk+BtOJRoIJFArC3NDGYSkjsOJJhLPt54g6HGJpqgFn5mhmQLwzN38ixnpxVbMBxLy20gwpaDNxLYnhUz3M4xk2BsOExYi8FhxgaJA2yHExtuniFaCzOTZGPb4cQNN3iI1CLZzMZszHAmzViyB+iXBGL8ws9//uNjhgobOX72wxtvfKixIawFxZFERw2SFlJ1jIJRMApGwcgAANTIQQRkI8lWAAAAAElFTkSuQmCC","orcid":"","institution":"Shanghai First Maternity and Infant Hospital","correspondingAuthor":true,"prefix":"","firstName":"Huifen","middleName":"","lastName":"Chen","suffix":""}],"badges":[],"createdAt":"2024-04-22 07:20:31","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4303861/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4303861/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":56397497,"identity":"c449dc47-9c18-4fa7-8a08-418da07f87e7","added_by":"auto","created_at":"2024-05-13 15:50:12","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":116371,"visible":true,"origin":"","legend":"\u003cp\u003eIdentification of deregulated circular RNAs (circRNAs) in gastric cancer (GC) tissues. (\u003cstrong\u003eA\u003c/strong\u003e) Clustered heatmap. Each row represents a circRNA, and each column represents a tissue sample. The color scale reflects the log2 signal strength from green (low intensity) to black (medium intensity) to red (strong intensity). (\u003cstrong\u003eB\u003c/strong\u003e) Volcano plots. The red points in plot indicate the differentially upregulated expression of circRNAs with statistical significance while the green points indicate the downregulated circRNAs.\u003c/p\u003e","description":"","filename":"F1.png","url":"https://assets-eu.researchsquare.com/files/rs-4303861/v1/22c02769888577a0efb96171.png"},{"id":56397496,"identity":"fa0ada86-5cce-4ef3-af12-d682ea0e29f4","added_by":"auto","created_at":"2024-05-13 15:50:12","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":36038,"visible":true,"origin":"","legend":"\u003cp\u003eThe relative expression levels of plasma circ_0007167 in gastric cancer group, benign gastric lesion group and healthy control group.\u003c/p\u003e","description":"","filename":"Picture1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4303861/v1/bfcb1c8fee591bba66a1ab98.jpg"},{"id":56397498,"identity":"78e7b356-bbd3-40ed-b570-f0da760f2134","added_by":"auto","created_at":"2024-05-13 15:50:12","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":28287,"visible":true,"origin":"","legend":"\u003cp\u003eThe diagnostic efficiency of circ_0007167 and CA19-9, CEA in GC detection.\u003c/p\u003e","description":"","filename":"Picture2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4303861/v1/1776ae42c8e9c246d480bd67.jpg"},{"id":56397500,"identity":"69f5109a-026f-4442-ad16-23c42b7cad5e","added_by":"auto","created_at":"2024-05-13 15:50:12","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":30501,"visible":true,"origin":"","legend":"\u003cp\u003eThe diagnostic efficiency of circ_0007167 and CA19-9, CEA in GC detection.\u003c/p\u003e","description":"","filename":"Picture3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4303861/v1/1483470cac40a0f8e7b01298.jpg"},{"id":56397501,"identity":"ba390cd0-d01e-458c-ac18-382e748d5bbd","added_by":"auto","created_at":"2024-05-13 15:50:12","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":29712,"visible":true,"origin":"","legend":"\u003cp\u003eDynamic monitoring of plasma circ_0007167 in gastric cancer patients before and after surgery\u003c/p\u003e","description":"","filename":"Picture4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4303861/v1/7ae3369ab74dc4c6164c1f18.jpg"},{"id":62286062,"identity":"a0799c69-7ad7-47a5-b4f5-253be01fbeb0","added_by":"auto","created_at":"2024-08-12 13:30:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":938673,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4303861/v1/a1c13848-b547-4723-9cc8-1fd57c201440.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"The expression and clinical significance of circ_0007167 in patients with gastric cancer","fulltext":[{"header":"1 Introduction","content":"\u003cp\u003eGastric cancer (GC) is a malignant tumor that originates from the epithelium of the stomach mucosa, ranking fifth in global cancer incidence rates [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e], and most of its occurrences are found in East Asia [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The main risk factors for GC are Helicobacter pylori infection, while other risk factors include geographical location, age, gender, smoking, salted meat consumption and genetics [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. GC is often asymptomatic in the early stages, and about 80% of patients have no obvious symptoms at the early stage of the disease, leading to easy missed diagnosis at the early stage [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Due to the lack of reliable and effective early diagnosis techniques, many patients are still diagnosed at an advanced stage, resulting in a 5-year survival rate of only 30% [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eCircular RNA (circRNA) was first discovered in the 1970s [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. More than 40 years ago, people first observed that eukaryotic RNA can exist in a circular form using electron microscopy [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. CircRNA is a covalently closed circular structure formed by the direct connection of the 5' and 3' ends, without a 3' tail or a 5' cap structure [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. In recent years, more and more studies have found that circRNA is closely related to the development of GC. Liu found that the expression level of circYAP1 in GC tissues was significantly lower than that in adjacent tissues, and the survival time of patients with low expression of circYAP1 was shorter than that of patients with high expression of circYAP1. CircYAP1 exerts tumor suppressor effects in GC cells by targeting the miR-367-5p/p27 axis [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Jie found that the expression of circMRPS35 in GC tissues was significantly down-regulated, which was associated with tumor size, TNM stage, and lymphatic metastasis, and was positively correlated with the expression of FOXO1/3a [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOver the past few decades, the global incidence and mortality rates of GC have been decreasing, which is closely related to the decline in Helicobacter pylori infection rates, as well as improvements in hygiene conditions and the widespread use of antibiotics [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Although the incidence is decreasing, the burden of GC patients worldwide will increase in the coming years due to the continuous growth and aging of the world's population [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. The 5-year survival rate is usually higher in patients with early GC (\u0026gt;\u0026thinsp;60%), while patients with GC with local and distant metastases have a significantly lower 5-year survival rate. Due to the atypical and occult nature of early GC, more than 60% of patients are already in the advanced stage at diagnosis [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. However, commonly used biomarkers for early GC screening, such as carcinoembryonic antigen (CEA) and carbohydrate antigen 19\u0026thinsp;\u0026minus;\u0026thinsp;9 (CA19-9), have low sensitivity and specificity rates [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Currently, endoscopic biopsy combined with pathological examination remains the gold standard for GC diagnosis, but its invasive nature and high cost limit its clinical acceptance [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Therefore, the low rate of early diagnosis is the main reason for poor prognosis in GC, so there is an urgent need to find non-invasive detection methods and biomarkers with high sensitivity and specificity to improve early diagnosis and survival rates. Based on the above theoretical foundation, this study aims to explore the expression and clinical significance of circ_0007167 in GC.\u003c/p\u003e"},{"header":"2 Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Subjects\u003c/h2\u003e \u003cp\u003eOne hundred and eight plasma samples were collected from GC patients in the affiliated hospital of Nantong University. All patients had not received radiotherapy, chemotherapy or other treatment before hospitalization. Forty patients with benign gastric lesions and 79 healthy subjects were also recruited in the affiliated hospital of Nantong University. All plasma samples were the leftover samples from routine medical tests in the Department of Medical Examination. All subjects in this study were reviewed and approved by the Medical Ethics Committee of the affiliated hospital of Nantong University, and the subjects provided informed consent.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2circRNA Sequencing\u003c/h2\u003e \u003cp\u003eThrough tissue-based circRNA sequencing, we identified 815 circRNAs with a fold change greater than 2 and p\u0026lt;0.05. By comparing the circRNA sequencing results of three additional pairs of tissues in the GEO database, we intersected them to obtain 73 circRNAs molecules that were consistently up-regulated across different sequencing datasets. By analyzing the correlation between module feature genes and clinical traits, we identified functionally distinct modules. We selected the module positively associated with GC carcinogenesis and intersected it with the two sequencing datasets, leading us to the identification of circRNA_0007167 as a candidate involved in GC development and exhibiting significant up-regulation for further investigation.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Realtime PCR and determination of CA19-9, CEA levels\u003c/h2\u003e \u003cp\u003eTotal RNA was extracted from cells using TRIzol reagent (Thermo Fisher Scientifc, California, USA). A PrimeScript RT Reagent Kit (Thermo Fisher Scientifc, California, USA) was used for RNA generation. Quantitative real-time PCR (qRT-PCR) was performed by a SYBR Premix Ex Taq II (TaKaRa, Japan) via a 7900HT real-time PCR system (Thermo Fisher Scientifc, California, USA). The PCR reaction uses 18S rRNA as an internal reference and employs the 2-ΔΔct quantitative method. Primers are listed as below:\u003c/p\u003e \u003cp\u003ecirc_0007167-F: 5'-TACCGTCAAATCCCAAGCCC-3'\u003c/p\u003e \u003cp\u003ecirc_0007167-R: 5'-AGACATGAGCAGACCAGAATCT-3'\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003e18s-F: 5'-CGGCTACCACATCCAAGGAA-3'\u003c/h3\u003e\n\n\u003ch3\u003e18s-R: 5'-GCTGGAATTACCGCGGCT-3'\u003c/h3\u003e\n\u003cp\u003eDetection of CA19-9 and CEA levels using Abbott i2000 chemiluminescence analyzer (Abbott /USA) in strict accordance with the instructions for use.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Statistical analyses\u003c/h2\u003e \u003cp\u003eStatistical analyses were performed by the SPSS 21.0 (Armonk, USA) and GraphPad Prism 8.0 (San Diegl, USA) software. The relative expression of circRNA_0007167 in plasma and the contents of CEA and CA19-9 in different groups were expressed by the median (interquartile range) [M (P25, P75)]. The differences in the expression levels of the three groups were compared by one-way analysis of variance, and pairwise comparisons were performed using the LSD method. The categorical variables were expressed by number (percentage) and compared between groups using the χ2 test. The diagnostic value of individual and combined diagnostic indicators in clinical diagnosis of gastric cancer was evaluated using the receiver operating characteristic (ROC) curve and the 95% confidence interval (95%CI) of the area under the curve (AUC). The comparison of the AUC areas was performed using paired Z-tests. The comparison of diagnostic evaluation indicators (accuracy, specificity, sensitivity, positive predictive value, and negative predictive value) between groups was performed using chi-square tests and Boffernoni multiple comparisons. \u003cem\u003eP\u003c/em\u003e values less than 0.05 were considered statistically significant (*, p\u0026lt;0.05;**, p\u0026lt;0.01༛***, p\u0026lt;0.001).\u003c/p\u003e \u003c/div\u003e"},{"header":"3 Results","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Identification of Deregulated circRNA in GC Tissues\u003c/h2\u003e \u003cp\u003eTo investigate the expression profiles of circRNAs in GC tissues, we conducted high-throughput sequencing in three GC tissues vs. three matched noncancerous tissues and identified a total of 25,303 circRNA targets, including 20,036 known circRNAs and 5,267 undefined circRNAs. The heatmap was depicted as a direct approach to visualize the distributions of the dataset for circRNA profiles (Fig.\u0026nbsp;1A). Volcano plots depicted 2,007 differentially expressed circRNAs in the GC tissues (Flod Change\u0026gt;2.5, p\u0026lt;0.05) (Fig.\u0026nbsp;1B). To identify the target circRNA, we searched the circRNA database for their chromosomal locations, transcript information, ORF, IRES, and other information. We also read the literature and selected five circRNAs for PCR verification. In both tissues and plasma, circ_0007167 showed significant differences and high correlation, so we ultimately selected circ_0007167 as the target molecule.\u003c/p\u003e \u003cp\u003e \u003cb\u003e(A)\u003c/b\u003e (\u003cb\u003eB)\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eFigure 1\u003c/strong\u003e \u003cp\u003eIdentification of deregulated circular RNAs (circRNAs) in gastric cancer (GC) tissues. (\u003cb\u003eA\u003c/b\u003e) Clustered heatmap. Each row represents a circRNA, and each column represents a tissue sample. The color scale reflects the log2 signal strength from green (low intensity) to black (medium intensity) to red (strong intensity). (\u003cb\u003eB\u003c/b\u003e) Volcano plots. The red points in plot indicate the differentially upregulated expression of circRNAs with statistical significance while the green points indicate the downregulated circRNAs.\u003c/p\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Detection of relative expression level of plasma circ_0007167\u003c/h2\u003e \u003cp\u003e108 plasma samples were collected from gastric cancer patients, including 70 males and 38 females, aged from 36 to 85 years old. 40 plasma samples were collected from patients with benign gastric lesions, including 25 males and 15 females, aged from 43 to 85 years old. 79 plasma samples were collected from healthy subjects, including 42 males and 37 females, aged from 40 to 89 years old. Intergroup comparison showed that the genders were matched (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\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\u003eComparison of the gender composition of the study subjects\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGender\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGC(\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;108)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGastric benign lesions(\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;40)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHealthy controls(\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;79)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003emale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e70(64.81)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e25(62.50)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e42(53.16)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003efemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e38(35.19)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15(37.50)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e37(46.84)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eχ\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.682\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ep-Value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.262\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe relative expression levels of plasma circ_0007167 were measured by qRT-PCR in 108 gastric cancer patients, 40 patients with benign gastric lesions, and 79 healthy subjects. The data were processed using SPSS 21.0. The relative expression levels of plasma circ_0007167 in the gastric cancer group, benign gastric lesion group, and healthy control group were 5.174 (2.741, 7.898), 1.641 (1.341, 2.086), and 1.027 (0.724, 1.561), respectively. The results of multiple comparisons using analysis of variance showed that the relative expression levels of plasma circ_0007167 were significantly higher in the gastric cancer group than in the benign gastric lesion group and healthy control group (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). There was no significant difference in the relative expression levels of plasma circ_0007167 between the benign gastric lesion group and healthy control group (p\u0026thinsp;=\u0026thinsp;0.328) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Correlation analysis of circ_0007167 expression and the clinicopathological parameters in GC patients\u003c/h2\u003e \u003cp\u003eAs shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, the expression of circ_0007167 in GC tissues was significantly correlated with tumor size (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05), T staging (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), lymph node metastasis (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), and nerve/vascular invasion (p\u0026thinsp;=\u0026thinsp;0.006). However, we did not find any association between the circ_0007167 expression and other clinicopathological parameters, such as gender (p\u0026thinsp;=\u0026thinsp;0.687), age (p\u0026thinsp;=\u0026thinsp;0.841), and differentiation degree (p\u0026thinsp;=\u0026thinsp;0.699).\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\u003eThe association between circ_0007167 expression and the clinicopathological parameters in GC patients.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCharacteristics\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHigh expression\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;108)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLow expression\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;54)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eχ\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\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\u003eGender\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003emale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e34(48.57)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36(51.43)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.162\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.687\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003efemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e20(52.63)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e18(47.37)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge(years)\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026lt;60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e20(51.28)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e19(48.72)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.040\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.841\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026ge;\u0026thinsp;60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e34(49.28)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e35(50.72)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTumor size(cm)\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026lt;5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e29(42.65)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e39(57.35)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.971\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.046*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026ge;\u0026thinsp;5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e25(62.50)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e15(37.50)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDegree of differentiation\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHigh-moderate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e31(51.67)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e29(48.33)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.150\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.699\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLow\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e23(47.92)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e25(52.08)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTumor depth\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 \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT1-T2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e42(75.00)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e14(25.00)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e29.077\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;0.001***\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT3-T4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e12(23.08)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e40(76.92)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLymph node metastasis\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 \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\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e38(79.17)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10(20.93)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e29.400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u0026lt;0.001***\u003c/p\u003e \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\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e16(26.67)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e44(73.33)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNerve/vascular 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 \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\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e28(66.67)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e14(33.33)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e7.636\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.006\u003c/p\u003e \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\u003e66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26(39.39)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e40(60.61)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.4Evaluation of diagnostic performance of plasma circ_0007167, CA19-9, and CEA\u003c/h2\u003e \u003cp\u003eBased on the expression levels of plasma circ_0007167, CA19-9, and CEA in 108 GC patients and 79 healthy individuals, ROC curves were plotted. It was found that the diagnostic performance of circ_0007167 was the best, with an area under the ROC curve of 0.881 (95% CI: 0.829\u0026ndash;0.943; p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). When CEA was used as a diagnostic indicator, the area under the ROC curve was 0.698 (95% CI: 0.620\u0026ndash;0.775; p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). When CA19-9 was used as a diagnostic indicator, the area under the ROC curve was 0.679 (95% CI: 0.601\u0026ndash;0.756; p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFurthermore, a pairwise comparison of the areas under the ROC curves showed that the diagnostic performance of circ_0007167 was significantly better than that of CEA or CA19-9 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001 for both comparisons). There was no significant difference in the diagnostic performance between CEA and CA19-9(p\u0026thinsp;\u003cem\u003e=\u003c/em\u003e\u0026thinsp;0.463)(Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of areas under the ROC curves for diagnosis using three indicators\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eComparison group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAUC difference\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eZ\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 \u003cp\u003ecirc_0007167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCEA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.184\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.209\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecirc_0007167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCA19-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.203\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.756\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCEA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCA19-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.019\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.463\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.463\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003e3.5 Evaluation of the diagnostic performance of combined detection of plasma circ_0007167, CA19-9, and CEA\u003c/b\u003e\u003c/h2\u003e \u003cp\u003eAfter combining circ_0007167 with CA19-9 and CEA for diagnosis and plotting ROC curves, it was found that when circ_0007167 was combined with CEA as a diagnostic indicator, the area under the ROC curve was 0.893 (95% CI: 0.843\u0026ndash;0.943; p\u0026thinsp;\u0026lt;\u0026thinsp;0.001); when circ_0007167 was combined with CA19-9 as a diagnostic indicator, the area under the ROC curve was 0.891 (95% CI: 0.842\u0026ndash;0.940; p\u0026thinsp;\u0026lt;\u0026thinsp;0.001); when circ_0007167, CEA, and CA19-9 were combined as diagnostic indicators, the area under the ROC curve was 0.896 (95% CI: 0.846\u0026ndash;0.945; p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFurther pairwise comparison of the areas under the ROC curves showed that there were no statistically significant differences between any of the three groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), indicating that the diagnostic efficacies of circ_0007167 combined with CEA or CA19-9, as well as all three combined, are equivalent (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of areas under the ROC curves for three combined diagnostic methods\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eComparison group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eAUC difference\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eZ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\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\u003ecirc_0007167\u0026thinsp;+\u0026thinsp;CEA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ecirc_0007167\u0026thinsp;+\u0026thinsp;CA19-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.002\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.193\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.847\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecirc_0007167\u0026thinsp;+\u0026thinsp;CEA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ecirc_000716\u0026thinsp;+\u0026thinsp;CEA\u0026thinsp;+\u0026thinsp;CA19-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-0.003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e-0.366\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.714\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecirc_0007167\u0026thinsp;+\u0026thinsp;CA19-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ecirc_000716\u0026thinsp;+\u0026thinsp;CEA\u0026thinsp;+\u0026thinsp;CA19-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-0.004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e-0.439\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.661\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eFurther analysis of the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of plasma circ_0007167, CEA, and CA19-9 as individual diagnostic indicators and combined indicators for GC patients showed that the sensitivity of circ_0007167 was 86.11%, which was superior to CEA (65.74%) and CA19-9 (50.93%). The sensitivity was improved by pairwise and triplet combinations, with the highest sensitivity of 92.59% achieved by the triplet combination.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEvaluation of the diagnostic values of combination of circ_0007167, CEA and CA19-9.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSEN(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSPE(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eACCU(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePPV(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNPV(%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecirc_0007167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e86.11\u003c/p\u003e \u003cp\u003e(93/108)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e87.34\u003c/p\u003e \u003cp\u003e(69/79)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e86.63\u003c/p\u003e \u003cp\u003e(162/187)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e90.29\u003c/p\u003e \u003cp\u003e(93/103)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e82.14\u003c/p\u003e \u003cp\u003e(69/84)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCEA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e65.74\u003c/p\u003e \u003cp\u003e(71/108)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e65.82\u003c/p\u003e \u003cp\u003e(52/79)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e65.78\u003c/p\u003e \u003cp\u003e(123/187)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e72.45\u003c/p\u003e \u003cp\u003e(71/98)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e58.43\u003c/p\u003e \u003cp\u003e(52/89)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCA19-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50.93\u003c/p\u003e \u003cp\u003e(55/108)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e74.68\u003c/p\u003e \u003cp\u003e(59/79)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e60.96\u003c/p\u003e \u003cp\u003e(114/187)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e73.33\u003c/p\u003e \u003cp\u003e(55/75)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e52.68\u003c/p\u003e \u003cp\u003e(59/112)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecirc_0007167\u0026thinsp;+\u0026thinsp;CEA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e91.67\u003c/p\u003e \u003cp\u003e(99/108)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e49.37\u003c/p\u003e \u003cp\u003e(39/79)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e73.80\u003c/p\u003e \u003cp\u003e(138/187)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e71.22\u003c/p\u003e \u003cp\u003e(99/139)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e81.25\u003c/p\u003e \u003cp\u003e(39/48)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecirc_0007167\u0026thinsp;+\u0026thinsp;CA19-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e87.04\u003c/p\u003e \u003cp\u003e(94/108)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55.70\u003c/p\u003e \u003cp\u003e(44/79)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e73.80\u003c/p\u003e \u003cp\u003e(138/187)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e72.87\u003c/p\u003e \u003cp\u003e(94/129)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e75.86\u003c/p\u003e \u003cp\u003e(44/58)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecirc_0007167\u0026thinsp;+\u0026thinsp;CEA\u0026thinsp;+\u0026thinsp;CA19-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e92.59\u003c/p\u003e \u003cp\u003e(100/108)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e37.97\u003c/p\u003e \u003cp\u003e(30/79)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e69.52\u003c/p\u003e \u003cp\u003e(130/187)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e67.11\u003c/p\u003e \u003cp\u003e(100/149)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e78.95\u003c/p\u003e \u003cp\u003e(30/38)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eSEN, sensitivity; SPE, specificity; ACCU, overall accuracy; PPV, positive predictive value; NPV, negative predictive value\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e3.6 Monitoring ability of plasma circ_0007167 for GC disease progression\u003c/h2\u003e \u003cp\u003eTo dynamically detect the expression level of circ_0007167 in the plasma of 20 patients newly diagnosed with GC, and to analyze the differences between the two groups using paired t-test analysis. The results showed that the plasma circ_0007167 level decreased overall in GC patients after surgery and was statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4 Discussion","content":"\u003cp\u003eGastric cancer (GC) is one of the most common cancers worldwide, with over 1\u0026nbsp;million new cases reported in 2018. Nearly two-thirds of these cases occur in developing countries, especially in East Asia, where the incidence rate is the highest. The risk of GC is 2\u0026ndash;3 times higher in men than in women, and the risk increases with age, with the highest incidence rate occurring between the ages of 60 and 70 years old [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Due to the asymptomatic nature of early GC, patients often present with symptoms such as weight loss and abnormal pain by the time they are diagnosed with advanced GC, leading to a poor overall survival rate (OS) of less than 30% [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. However, in Japan and Korea, large-scale screening has increased the early diagnosis rate of GC, resulting in an OS of over 60% [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Therefore, early detection, diagnosis, and treatment of GC may be the most effective way to address this issue. It is crucial to identify new potential diagnostic biomarkers and therapeutic targets to improve the diagnosis and treatment of GC [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. This study aims to identify circular RNA markers associated with GC that may contribute to its diagnosis and treatment.\u003c/p\u003e \u003cp\u003eIn recent years, circRNA has become a research hotspot in the diagnosis and treatment of GC. More and more high-throughput sequencing and microarray gene chip technologies have shown significant differences in the expression levels of circRNA between GC patients and healthy individuals [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Additionally, the study of circRNA in peripheral blood has the advantages of convenient collection, minimal trauma, and high patient acceptance, making it a potential new biomarker for tumors.\u003c/p\u003e \u003cp\u003eIn view of the fact that there are few studies on GC diagnosis using serum or plasma circ_0007167 as biomarkers, this research group has selected circ_0007167 as the research object through gene chip screening in the early stage to explore its significance in GC auxiliary diagnosis. After statistical processing of the relative expression quantity of plasma circ_0007167 by 2\u003csup\u003e\u0026minus;△△CT\u003c/sup\u003e method, it was found that the content of plasma circ_0007167 in GC patients was significantly higher than that in patients with benign gastric lesions and healthy controls, while there was no significant difference between the benign gastric lesion group and the healthy control group. Further analysis of clinical pathological characteristics found that the level of plasma circ_0007167 was not related to patient's gender, age, and differentiation degree, but was related to tumor size, T staging, lymph node metastasis and nerve/vascular invasion. It shows that the level of plasma circ_0007167 is associated with the progression of GC patients to a certain extent.\u003c/p\u003e \u003cp\u003eROC curve analysis of diagnostic efficacy found that the area under the curve (AUC) of circ_0007167 was 0.881, significantly higher than that of CA19-9 and CEA. The diagnostic efficacy of circ_0007167 was better than that of traditional markers CEA and CA19-9. An AUC of 0.7\u0026ndash;0.9 suggests good clinical diagnostic value, while an AUC of 0.5\u0026ndash;0.7 indicates poor clinical diagnostic value [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Further pairwise comparison of the AUC showed that the diagnostic efficacy of circ_0007167 was higher than that of CEA or CA19-9, while there was no significant difference in the diagnostic efficacy between CEA and CA19-9. When circ_0007167 was combined with CA19-9 and CEA for diagnosis, the ROC curve showed that the AUC was 0.893 when circ_0007167 was combined with CEA, 0.891 when circ_0007167 was combined with CA19-9, and 0.896 when circ_0007167, CEA, and CA19-9 were combined as diagnostic indicators, with the best diagnostic efficacy. The sensitivity of circ_0007167 combined with CEA was 91.67%, and the specificity was 49.37%. The sensitivity of circ_0007167 combined with CA19-9 was 87.04%, and the specificity was 55.70%. The sensitivity of all three markers combined was 92.59%, and the specificity was 37.97%, with a decrease in specificity. Therefore, the combination of circ_0007167, CA19-9, and CEA can improve the diagnostic efficacy. Surgical resection is the main treatment for GC, but how to improve postoperative survival rates remains a hot topic [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. We found through simultaneous follow-up of GC patients that preoperative plasma circ_0007167 expression was higher, but it decreased postoperatively, suggesting that plasma circ_0007167 may have the ability to monitor GC progression.\u003c/p\u003e \u003cp\u003eIn summary, the high expression of plasma circ_0007167 in GC patients can be used as a new biomarker for GC diagnosis and prognostic monitoring. In the later stage, we will further expand the sample size and conduct follow-up studies to analyze the correlation between survival time and plasma circ_0007167 expression levels in GC patients, further validating the clinical application value of plasma circ_0007167 in GC patients. Peripheral blood tumor markers are important indicators reflecting tumor progression and prognosis in clinical practice [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. In the future, we will further study the biological function of circ_0007167 in GC, providing new ideas for the diagnosis and treatment of GC.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; Contributions:\u0026nbsp;\u003c/strong\u003eAll authors contributed to the conception and the main idea of the work. FLZ\u0026nbsp;wrote the manuscript and performed the experiences. FLZ, YTW, ZY, XTW, XPC and HFC analyzed the data and edited the manuscript. All authors reviewed the results and approved the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Approval and Informed Consent Statement:\u0026nbsp;\u003c/strong\u003eThe study was approved by the ethics committee of the Affiliated Hospital of Nantong University.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of Data and Materials:\u0026nbsp;\u003c/strong\u003eThe datasets generated for this study can be found in GEO database, GSE131414.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Statement\u003c/strong\u003e\u003cstrong\u003e:\u003c/strong\u003e This project was supported by grants from the National Natural Science Foundation of China (81871720，82072363).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest:\u0026nbsp;\u003c/strong\u003eThe authors declare that they have no conflicts of interest to report regarding the present study.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eFock KM. Review article: the epidemiology and prevention of gastric cancer. J Aliment Pharmacol Ther. 2014;40(3):250\u0026ndash;60.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDanaei G, Vander Hoorn S, Lopez AD, Murray CJ, Ezzati M. Causes of cancer in the world: comparative risk assessment of nine behavioural and environmental risk factors. J Lancet. 2005;366(9499):1784\u0026ndash;93.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLyons K, Le LC, Pham YT, Borron C, Park JY, Tran CTD, Tran TV, Tran HT, Vu KT, Do CD, et al. Gastric cancer: epidemiology, biology, and prevention: a mini review. J Eur J Cancer Prev. 2019;28(5):397\u0026ndash;412.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEusebi LH, Telese A, Marasco G, Bazzoli F, Zagari RM. Gastric cancer prevention strategies: a global perspective. J Gastroenterol Hepatol. 2020;35(9):1495\u0026ndash;502.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHashad D, Elbanna A, Ibrahim A, Khedr G. Evaluation of the role of circulating long non-coding RNA h19 as a promising novel biomarker in plasma of patients with gastric cancer. J Clin Lab Anal. 2016;30(6):1100\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKolakofsky D. Isolation and characterization of sendai virus di-RNAs. J Cell. 1976;8(4):547\u0026ndash;55.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHsu MT, Coca-Prados M. Electron microscopic evidence for the circular form of RNA in the cytoplasm of eukaryotic cells. J Nat. 1976;280(5720):339\u0026ndash;40.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePamudurti NR, Bartok O, Jens M, Ashwal-Fluss R, Stottmeister C, Ruhe L, Hanan M, Wyler E, Perez-Hernandez D, Ramberger E, et al. Translation of circRNAs. J Mol Cell. 2017;66(1):9\u0026ndash;21.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu H, Liu Y, Bian Z, Zhang J, Zhang R, Chen X, Huang Y, Wang Y, Zhu J. Correction to: circular RNA yap1 inhibits the proliferation and invasion of gastric cancer cells by regulating the mir-367-5p/p27 kip1 axis. J Mol Cancer. 2019;18(1):117.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJie M, Wu Y, Gao M, Li X, Liu C, Ouyang Q, Tang Q, Shan C, Lv Y, Zhang K, et al. CircMRPS35 suppresses gastric cancer progression via recruiting KAT7 to govern histone modification. J Mol Cancer. 2020;19(1):56.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFerro A, Peleteiro B, Malvezzi M, Bosetti C, Bertuccio P, Levi F, Negri E, La Vecchia C, Lunet N. Worldwide trends in gastric cancer mortality (1980\u0026ndash;2011), with predictions to 2015, and incidence by subtype. J Eur J Cancer. 2014;50(7):1330\u0026ndash;44.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFerlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. J Int J Cancer. 2013;136(5):E359\u0026ndash;86.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThrift AP, El-Serag HB. Burden of gastric cancer. J Clin Gastroenterol Hepatol. 2019;18(3):534\u0026ndash;42.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShimada H, Noie T, Ohashi M, Oba K, Takahashi Y. Clinical significance of serum tumor markers for gastric cancer: a systematic review of literature by the Task Force of the Japanese Gastric Cancer Association. J Gastric Cancer. 2014;17(1):26\u0026ndash;33.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSuzuki T, Kitagawa Y, Nankinzan R, Yamaguchi T. Early gastric cancer diagnostic ability of ultrathin endoscope loaded with laser light source. J World J Gastroenterol. 2019;25(11):94\u0026ndash;102.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJin Y, Park, Lawrence VK, Rolando. Herrero. Prevention strategies for gastric cancer: a global perspective. J Clin Endosc. 2014;47(6):478\u0026ndash;89.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAllemani C, Weir HK, Carreira H, Harewood R, Spika D, Wang XS, Bannon F, Ahn JV, Johnson CJ, Bonaventure A, et al. Global surveillance of cancer survival 1995\u0026ndash;2009: analysis of individual data for 25,676,887 patients from 279 population-based registries in 67 countries (CONCORD-2). J Lancet. 2015;385(9972):977\u0026ndash;1010.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMinicozzi P, Innos K, S\u0026aacute;nchez MJ, Trama A, Walsh PM, Marcos-Gragera R, Dimitrova N, Botta L, Visser O, Rossi S, et al. Quality analysis of population-based information on cancer stage at diagnosis across Europe, with presentation of stage-specific cancer survival estimates: A EUROCARE-5 study. J Eur J Cancer. 2017;84:335\u0026ndash;53.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWu W, Zhen T, Yu J, Yang Q. Circular RNAs as New Regulators in Gastric Cancer: Diagnosis and Cancer Therapy. J Front Oncol. 2020; 101526.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiang M, Liu Z, Lin H, Shi B, Li M, Chen T, Qin L, Niu Q, Yu G, Jiang H. High-throughput sequencing reveals circular RNA hsa_circ_0000592 as a novel player in the carcinogenesis of gastric carcinoma. J Biosci Rep. 2019; 39(6).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTang T, Fu W, Sun K, Rong H, Wang D, Cao H. CircRNA microarray profiling identifies a novel circulating biomarker for detection of gastric cancer. J Mol Cancer. 2018;17(1):137.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eToft JH, \u0026Oslash;kland I, Dalen I. ROC-curves-fundamentals for proper use. J Endocr. 2022;76(2):505.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAjani JA, Lee J, Sano T, Janjigian YY, Fan D, Song S. Gastric adenocarcinoma. J Nat Rev Dis Primers. 2017;3(2):17036.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSo JBY, Kapoor R, Zhu F, Koh C, Zhou L, Zou R, Tang YC, Goo PCK, Rha SY, Chung HC, et al. Development and validation of a serum microRNA biomarker panel for detecting gastric cancer in a high-risk population. J Gut. 2021;70(5):829\u0026ndash;37.\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":"circRNA, circ_0007167, gastric cancer, diagnosis","lastPublishedDoi":"10.21203/rs.3.rs-4303861/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4303861/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eGastric cancer (GC) is one of the commonest malignant tumors worldwide including China. Data from epidemiological surveys showed that the new cases of GC were globally increased by 1\u0026nbsp;million per year, and 800,000 people died from gastric cancer every year. In recent years, it has been found that cyclic RNA plays an important role in the occurrence of GC, and it may be used as an evaluation index for the diagnosis and prognosis of GC. This study was conducted to assess the clinical application value of circ_0007167 for diagnosis and prognosis of GC.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eGC group was collected from September 1, 2019 to November 30, 2020 at the Affiliated Hospital of Nantong University. There were 108 patients with GC confirmed by imaging and pathology, including 20 pairs of plasma from GC patients before and one week after surgery. There were 40 patients with benign gastric lesions and 79 healthy individuals from the physical examination center of the Affiliated Hospital of Nantong University. The expression levels of circ_0007167 in the plasma samples of the three groups were detected using qRT-PCR following validation of the method. The correlations between changes in plasma circ_0007167 levels and the clinical pathological parameters of GC patients were statistically analyzed. Meanwhile, the ROC curves of plasma circ_0007167, CEA and CA19-9 levels were analyzed to study the clinical value of single and combined detection in the diagnosis and prognosis of GC.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThe expression level of circ_0007167 in the plasma of patients with GC was significantly higher than that in the benign gastric lesions group and the healthy control group (both p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). There was no significant difference between benign gastric lesion group and healthy control group (p\u0026thinsp;=\u0026thinsp;0.328). Further clinicopathologic features showed that plasma circ_0007167 was not related to gender (p\u0026thinsp;=\u0026thinsp;0.687), age (p\u0026thinsp;=\u0026thinsp;0.841), and differentiation degree of tumor (p\u0026thinsp;=\u0026thinsp;0.9), but related to tumor size (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05), T stage (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), metastasis (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and nerve/vascular invasion (p\u0026thinsp;=\u0026thinsp;0.006). The diagnostic efficacy of ROC curve analysis showed that the area under the curve (AUC) of circ_0007167 was 0.881, significantly higher than that of CA19-9 (AUC\u0026thinsp;=\u0026thinsp;0.679) and CEA (AUC\u0026thinsp;=\u0026thinsp;0.698), and the diagnostic efficacy of circ_0007167 was better than that of traditional tumor markers CEA and CA19-9. There was no significant difference in diagnostic efficacy between CEA and CA19-9, but when circ_0007167 was combined with CEA, AUC was 0. 893; when circ_0007167 was combined with CA19-9, AUC was 0. 891; when circ_0007167 was combined with CEA and CA19-9 as diagnostic markers, diagnostic efficiency is the best with AUC\u0026thinsp;=\u0026thinsp;0.896. The expression of circ_0007167 in plasma of GC patients decreased significantly after operation compared to that before operation (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003ePlasma circ_0007167 in patients with GC is superior to traditional tumor markers CEA and CA19-9. Combined detection of circ_0007167 with CEA and CA19-9 can improve the assisted diagnosis of early GC. Moreover, the plasma circ_0007167 was decreased significantly following surgery, suggesting that plasma circ_0007167 may serve as a new biomarker for the diagnosis, treatment, and prognosis evaluation of GC.\u003c/p\u003e","manuscriptTitle":"The expression and clinical significance of circ_0007167 in patients with gastric cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-05-13 15:50:08","doi":"10.21203/rs.3.rs-4303861/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":"812c592f-70e7-4499-81ac-24fe3d08ba88","owner":[],"postedDate":"May 13th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-08-12T13:22:53+00:00","versionOfRecord":[],"versionCreatedAt":"2024-05-13 15:50:08","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4303861","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4303861","identity":"rs-4303861","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}