Relationships between CD147 expression, tumor-infiltrating lymphocytes, and oncostatin M in hepatocellular carcinoma

Relationships between CD147 expression, tumor-infiltrating lymphocytes, and oncostatin M in hepatocellular carcinoma | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Relationships between CD147 expression, tumor-infiltrating lymphocytes, and oncostatin M in hepatocellular carcinoma Yasuyuki Shigematsu, Hiroaki Kanda, Yu Takahashi, Kengo Takeuchi, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4418866/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 Purpose In hepatocellular carcinoma (HCC), CD147 expression contributes to tumor malignancy; however, its relationship with the tumor-immune microenvironment (TIME) remains unclear. This study aimed to elucidate the clinicopathological characteristics associated with CD147 expression in HCC and investigate its association with the TIME, specifically its association with tumor-infiltrating lymphocytes (TILs) and oncostatin M (OSM). Methods Using 397 HCC specimens from patients undergoing curative-intent resection, we assessed CD147 expression in tumor cells and quantified OSM-positive cells and various TILs (CD8 + , CD4 + , FOXP3 + , and CD20 + cells) in the TIME. Using tissue microarrays, these assessments were performed through immunohistochemical analysis. We investigated the associations between CD147 expression status, the density of OSM-positive cells, and the densities of various TILs. Results High CD147 expression, found in 332 specimens (83.6%), was associated with advanced clinical stage ( P = 0.0029), fibrosis ( P = 0.036), and higher densities of FOXP3 + cells ( P = 0.0039), CD4 + cells ( P = 0.0012), and OSM-positive cells ( P = 0.0017). In CD147-high tumors, OSM-positive cell density was associated with all assessed TIL subsets (CD8 + , CD4 + , FOXP3 + , and CD20 + cells; all P s < 0.001), whereas in CD147-low tumors, OSM-positive cell density was associated only with FOXP3 + cells ( P = 0.0004). Conclusions In HCC, CD147 expression is associated with an immunosuppressive TIME, characterized by increased FOXP3 + regulatory T cells and an association with OSM-positive cells. These results elucidate the potential mechanisms through which CD147 facilitates tumor immune evasion, suggesting the CD147-OSM axis as a promising target for therapeutic intervention in HCC. CD20 CD4 CD8 FOXP3 hepatic fibrosis immunity immunohistochemistry liver cancer regulatory T cells tumor-immune microenvironment Figures Figure 1 Figure 2 Figure 3 Introduction Primary liver cancer is the sixth most commonly diagnosed cancer and the third leading cause of cancer-related deaths globally. 1 Hepatocellular carcinoma (HCC), which constitutes 75–85% of all primary liver cancer cases, 2 generally has a poor prognosis, with a 5-year overall survival rate of approximately 16%. This poor prognosis has remained relatively unchanged over the last three decades, 3 despite advancements in diagnosis and treatment. There is a critical need for a deeper understanding of both tumor cells and the tumor-immune microenvironment (TIME) to develop effective HCC treatment strategies. Recent studies have highlighted the importance of CD147, a glycoprotein originally known as a regulator of matrix metalloproteinase (MMP) in various tumor types, including HCC. 4 – 6 The overexpression of CD147 in tumor cells has been identified as a predictor of high malignant potential and advanced clinical stage. 7 – 9 CD147 expression in tumor cells induces mesenchymal cells to produce MMPs, 10 , 11 which degrade the extracellular matrix and create a niche conducive to tumor invasion and metastasis. Through its interaction with the transforming growth factor-β (TGF-β) signaling pathway, CD147 substantially enhances tumor invasiveness by facilitating the epithelial-mesenchymal transition (EMT) of HCC cells. 12 However, its influence on the TIME remains inadequately explored. Given recent advancements in antitumor immunotherapy, elucidating the association between CD147 expression and the TIME is essential, as it could facilitate the development of effective therapeutic strategies. Oncostatin M (OSM), a pleiotropic inflammatory cytokine produced mainly by macrophages and Kupffer cells in the liver, 13 , 14 also promotes EMT and advances HCC progression. 15 , 16 The association between CD147 expression and OSM, and their collective effects on the TIME and immune modulation, remains insufficiently explored. This raises questions regarding the potential role of CD147 and OSM in modulating tumor behavior and immune evasion in HCC. This study aimed to elucidate the clinicopathological features associated with CD147 expression in HCC and investigate its relationship with the TIME, thereby providing a basis for novel therapeutic strategies for the management of HCC. Materials and Methods Patients and specimens Specimens from surgically resected HCC were obtained from patients at the Cancer Institute Hospital of the Japanese Foundation for Cancer Research in Tokyo, Japan, between January 1, 2005 and December 31, 2016. These patients underwent resection with curative intent for HCC. Patients with congenital and chronic autoimmune liver diseases were excluded. This study included a cohort of 397 patients diagnosed with HCC, selected on the basis of the availability of formalin-fixed, paraffin-embedded (FFPE) tissue specimens for analyses. Clinicopathological variables, including height, weight, sex, preoperative blood test results, and the clinical tumor-node-metastasis (TNM) stage, were extracted from the medical record database. Hepatic functionality was evaluated utilizing the albumin-bilirubin (ALBI) score system. 17 Tumor staging adhered to the 8th edition of the TNM classification by the Unio Internationalis Contra Cancrum. 18 Approval for the study protocol was obtained from the ethics committee at the Japanese Foundation for Cancer Research (approval number 2017-GA-1136). The requirement for informed consent was waived because of the retrospective study design. Pathological evaluation Hematoxylin and eosin-stained 4 µm sections of FFPE samples from the patients with HCC were reviewed histologically by two experienced liver pathologists (Y.S. and H.K.). The diagnosis of HCC was performed according to the 5th edition of the World Health Organization criteria. 19 Pathological features were evaluated according to the Japanese general rules for clinical and pathological study of primary liver cancer. 20 A whole section of noncancerous liver tissues without any tumor cells was used to evaluate the background liver tissue of each patient. Hepatic fibrosis was evaluated based on the Brunt criteria. 21 Immunohistochemistry Tissue microarrays (TMAs) were prepared using FFPE tumor tissues to assess the immunohistochemical features of tumor cells and tumor-infiltrating lymphocytes (TILs), as previously described. 22 , 23 Briefly, designated tissue cores of the donor paraffin blocks were extracted using a coring needle measuring 2 mm in diameter. Subsequently, the tissue cores were arranged in an array on the recipient block using a KIN-1 manual tissue arrayer (Azumaya, Tokyo, Japan). Three tissue cores, representing the primary histology of each tumor, were selected. TMA sections of 4 µm thick were then used for subsequent immunohistochemical evaluations. Immunohistochemical evaluation of the tumor tissues was performed using anti-CD147 rabbit monoclonal antibody (1:60; clone E1S1V; Cell Signaling Technology, Inc., Danvers, MA, USA) and anti-oncostatin M rabbit polyclonal antibody (1:300; ab198830; Abcam, Cambridge, UK). TILs were assessed using anti-CD8 mouse monoclonal antibody (1:3; clone C8/144B; Nichirei, Tokyo, Japan), anti-CD4 mouse monoclonal antibody (1:2; clone 4B12; Nichirei), anti-CD20 mouse monoclonal antibody (1:800; clone L26; Leica, Wetzlar, Germany), and anti-FOXP3 mouse monoclonal antibody (1:100; clone 236A/E7; Abcam). For positive and negative controls for CD147, we used a cell array obtained from Daiichi Sankyo Co., Ltd., consisting of A431 (Cat #CRL-1555, ATCC, high expression), BxPC-3 (Cat #CRL-1687, ATCC, high expression), HCC1954 (Cat #CRL-2338, ATCC, high expression), ZR-75-1 (Cat #CRL-1500, ATCC, middle expression), MCF-7 (Cat #HTB-22, ATCC, low-middle expression), MDA-MB-453 (Cat #HTB-131, ATCC, low expression), and CHO-K1 cell lines (Cat #CCL-61, ATCC, negative). Cultured cell lines were collected and were fixed with 20% formalin neutral buffer solution (Cat. No. 060-01721, FUJIFILM Wako Pure Chemical Corporation), and were embedded with Histogel (Cat. No. HG-4000-012, Epredia Netherlands B.V.). These blocks were then replaced with paraffin and the slides of thin sections were prepared at Daiichi Sankyo Co., Ltd. Non-cancerous liver tissue was used for positive and negative controls for OSM, CD8, CD4, FOXP3, and CD20. Immunostaining was performed using a Bond-III automated staining system (Leica Microsystems, Buffalo Grove, IL, USA), and antigens were detected using a Bond Polymer Refine Detection Kit (Leica Microsystems). Evaluation of protein expression through immunohistochemistry in TMA CD147 expression was regarded as positive if circumferential membrane expression was present. CD147 expression in tumor tissues was categorized as high or low, according to the predominant expression pattern observed across the tumor cells, and HCC samples were classified into two groups based on the CD147 expression status: HCC showing high CD147 expression (CD147-high HCC) and HCC showing low CD147 expression (CD147-low HCC). To evaluate the number of OSM-positive cells and TILs, immunostained sections were scanned at ×40 magnification using a NanoZoomer Digital Pathology System (Hamamatsu Photonics KK, Shizuoka, Japan) at a resolution of 0.55 pixel/µm and semi-automatically digitized using the proprietary NanoZoomer Digital Pathology Image file format. TILs were counted using Fiji, an open-source platform for biological-image analysis. 24 A microscopic field was selected for each tissue core from the scanned TMA images. The mean number of TILs counted in the three tissue cores of each tumor was considered the number of TILs in the tumor. Statistical analysis Statistical analyses were performed using R version 4.2.0 (R Foundation for Statistical Computing, Vienna, Austria). Continuous variables were reported as means, medians, standard deviations, or standard errors as appropriate. Categorical variables were reported as numbers and percentages. The statistical significance of between-group differences of continuous variables was assessed using either the Mann-Whitney U test or Student’s t -test, and categorical variables were compared using the Fisher’s exact test. A two-tailed P value < 0.05 was considered statistically significant. Results CD147 expression and clinicopathological features The CD147 staining in the HCC specimens was relatively homogenous (Fig. 1 ). Of the 397 HCC tissue samples analyzed, 332 (83.6%) showed high CD147 expression. The clinicopathological features are summarized according to CD147 expression status in Table 1 . High CD147 expression was associated with advanced clinical stages ( P = 0.0029). A fibrous capsule or septal fibrosis was more frequently present in CD147-high HCC than in CD147-low HCC (94.0% vs. 86.2%, P = 0.036). No significant differences in CD147 status were observed according to age, sex, history of hepatitis virus infection, tumor differentiation, or presence of background liver cirrhosis. Table 1 Clinicopathological features of patients with HCC according to CD147 expression status. Variable Total (n = 397) CD147 P -value Low (n = 65) High (n = 332) Age (years) † 67.9 (± 10.1) 66.4 (± 10.9) 68.1 (± 9.9) 0.31 Sex § 0.57 Female 90 (22.7) 17 (26.2) 73 (22.0) Male 307 (77.3) 48 (73.8) 259 (78.0) BMI § 0.24 < 25 273 (68.8) 49 (75.4) 224 (67.5) 25 ≦ 124 (31.2) 16 (24.6) 108 (32.5) Hepatitis virus infection § 0.94 HBV 99 (24.9) 18 (27.7) 81 (24.4) HCV 138 (34.8) 21 (32.3) 117 (35.2) HBV + HCV 11 (2.8) 2 (3.1) 9 (2.7) Other 149 (37.5) 24 (36.9) 125 (37.7) ALBI § 0.11 Grade 1 210 (52.9) 27 (41.5) 183 (55.1) Grade 2 184 (46.3) 37 (56.9) 147 (44.3) Grade 3 3 (0.8) 1 (1.5) 2 (0.6) TNM stage § 0.0029 IA 32 (8.1) 8 (12.3) 24 (7.2) IB 52 (13.1) 4 (6.2) 48 (14.5) II 258 (65.0) 46 (70.8) 212 (63.9) IIIA 34 (8.5) 7 (10.8) 27 (8.1) IIIB 21 (5.3) 0 (0.0) 21 (6.3) Tumor differentiation § 0.59 Well 36 (9.0) 4 (6.2) 32 (9.6) Moderately 281 (70.8) 49 (75.2) 232 (69.9) Poorly 80 (20.2) 12 (18.5) 68 (20.5) Growth pattern § 0.81 Expansive 363 (91.4) 59 (90.8) 304 (91.5) Infiltrative 34 (8.6) 6 (9.2) 28 (8.5) Fibrous capsule or septal fibrosis in the tumor § 0.036 Absent 29 (7.3) 9 (13.8) 20 (6.0) Present 368 (92.7) 56 (86.2) 312 (94.0) Background liver cirrhosis § 1.00 Absent 235 (59.2) 39 (60.0) 196 (59.0) Present 162 (40.8) 26 (40.0) 136 (41.0) § Data presented as n (%); † Data presented as mean (± standard deviation). Abbreviations: ALBI, albumin-bilirubin; BMI, body mass index; HBV, hepatitis B virus; HCC, hepatocellular carcinoma; HCV, hepatitis C virus; TNM, tumor-node-metastasis. CD147 expression and tumor-infiltrating lymphocytes To evaluate the TIME associated with CD147 expression on tumor cells, we quantified densities of CD8 + , CD4 + , FOXP3 + , and CD20 + cells within the tumor tissues ( Fig. S1 ). Compared with CD147-low HCC, CD147-high HCC contained higher densities of CD4 + cells (median, 46.0/mm 2 vs. 104.5/mm 2 for CD147-low vs. CD147-high, respectively; P = 0.0012) and FOXP3 + cells (median, 11.0/mm 2 vs. 17.5/mm 2 for CD147-low HCC vs. CD147-high HCC, respectively; P = 0.0039) in the tumor tissues (Table 2 ). The density of CD8 + cells and CD20 + cells did not differ significantly according to the CD147 expression status of the tumor. These observations suggest that CD147 expression on tumor cells may influence TIME through the induction of regulatory T cells (Tregs). Therefore, in HCC, CD147 expression is associated with an immunosuppressive TIME. Table 2 Tumor infiltrating lymphocytes and CD147 in hepatocellular carcinoma. Type of cell CD147 P -value Low High CD8 + 127.0 (48.5-178.3) 125.5 (34.8-399.3) 0.25 CD4 + 46.0 (7.8–114.0) 104.5 (22.0-252.5) 0.0012 FOXP3 + 11.0 (6.0-30.5) 17.5 (8.0-67.3) 0.0039 CD20 + 9.5 (5.0-79.3) 14.0 (6.0-112.0) 0.27 Data presented as mean (1st quartile-3rd quartile). CD147 expression on tumor cells and OSM-positive cells CD147 is implicated in enhancing the invasiveness of hepatocytes by inducing EMT via the TGF-β signaling pathway. 12 Similarly, OSM is recognized for its role in promoting EMT in HCC. 25 To assess the association between CD147 and OSM, we evaluated their expression profiles using immunohistochemistry. OSM was predominantly expressed in stromal cells adjacent to the periphery of the tumor cell cords (Fig. 2 A). Immunohistochemical analysis revealed a higher density of OSM-positive cells in CD147-high than CD147-low HCC (median, 61/mm 2 vs. 25/mm 2 ; P = 0.0017; Fig. 2 B), suggesting possible interaction between CD147-expressing tumor cells and OSM-positive cells. Association between OSM-positive cells and FOXP3 + cells according to CD147 expression To assess the effect of OSM on TILs, we investigated the association of OSM-positive cells with CD8 + , CD4 + , FOXP3 + , and CD20 + cells ( Fig. S2 ). CD8 + , CD4 + , FOXP3 + , and CD20 + cells were significantly associated with OSM-positive cells, with correlation coefficients of 0.21, 0.34, 0.42, and 0.17, respectively (all P s < 0.001; Fig. 3 A). To evaluate the influence of CD147 expression on the associations between OSM and TILs, we stratified the analysis by CD147 expression status. In CD147-high HCC, CD8 + , CD4 + , FOXP3 + , and CD20 + were significantly associated with OSM-positive cells, with correlation coefficients of 0.22, 0.35, 0.41, and 0.18, respectively, each demonstrating a significant correlation (all P s < 0.001; Fig. 3 B). In contrast, in CD147-low HCC, the correlation coefficients for CD8 + , CD4 + , FOXP3 + , and CD20 + cells were − 0.070, 0.023, 0.431, and − 0.078, respectively, with only FOXP3 + cells showing a statistically significant correlation ( P = 0.0004; Fig. 3 C), suggesting that the infiltration of FOXP3 + cells in tumor tissues may be indirectly mediated through OSM rather than a direct effect of CD147. Discussion This study elucidates the potential role of CD147 in HCC, particularly its association with the TIME and its interaction with TILs and OSM. The results demonstrate that high CD147 expression, present in most HCC samples, is significantly associated with advanced disease stages and increased fibrosis, highlighting its potential as a biomarker of aggressive HCC phenotypes. Moreover, the observed increase in FOXP3 + Tregs in CD147-high HCC tumors suggests a potential role of CD147 in creating an immunosuppressive environment conducive to tumor progression and evasion from immune surveillance. Notably, the observed association between CD147 expression, OSM-positive cells, and FOXP3 + cells indicates a complex interplay that might contribute to an immunosuppressive TIME, highlighting the importance of the CD147-OSM axis in the modulation of immune responses within the TIME in HCC. The association between CD147 expression and TIME in HCC, particularly through the modulation of FOXP3 + and CD8 + TILs, illuminates the complex immunological dynamics influenced by CD147. The prognostic significance of these TIL subsets in HCC is well-documented: high CD8 + TIL counts are associated with more favorable prognosis, signifying effective anti-tumor immunity, whereas high FOXP3 + TIL counts, indicative of Tregs, are associated with a worse prognosis, reflecting an immunosuppressive milieu that facilitates tumor growth and immune escape. 26 Our findings, indicating pronounced FOXP3 + cell infiltration in CD147-high HCC tumors, emphasize a skewed balance towards an immunosuppressive state, potentially overriding the antitumor effects of CD8 + cytotoxic T cells. The observed interaction between CD147 and OSM indicates a potential synergistic contribution to EMT, thereby influencing tumor invasiveness and metastasis. CD147 is known to facilitate EMT via the TGF-β signaling pathway, 27 whereas OSM activates STAT3, which also triggers EMT in cancer cells. 15 OSM-activated STAT3 cooperates with TGF-β to induce mesenchymal stem cell properties in breast cancer and pancreatic cancer. 28 , 29 Although direct evidence of CD147 and OSM interaction in HCC has not been established, the correlation between high CD147 expression and increased OSM-positive cell density in the TIME suggest a cooperative interaction that may promote EMT. This relationship highlights a complex interplay of factors determining HCC progression and suggests novel potential therapeutic targets. The observed positive correlation between OSM-positive cells and FOXP3 + cells suggests that OSM may promote immunosuppression by Treg induction. This relationship highlights the complex relationship between cytokine signals and the regulation of the immune system within the TIME. Given the strong association between the density of OSM-positive cells in TIME and CD147 expression in tumor cells, the increased infiltration of FOXP3 + cells in CD147-high HCC may be influenced by OSM. Although the exact mechanism by which OSM increases the number of FOXP3 + Tregs remains elusive, existing knowledge suggests that OSM may indirectly contribute to the expansion of Tregs by inhibiting IL-17/Th17 activity in CD4 + T cells. 30 This study has several limitations. First, the retrospective nature of this study and restricting the study population to patients undergoing hepatectomy may have introduced inherent bias. Consecutive patients were included to mitigate sampling bias. Second, although the TMA method facilitated the immunohistochemical assessment of both tumor cells and the TIME, it might not have fully captured the heterogeneity of the entire tumor. To make the results more representative of the tumors, we carefully selected three tissue cores per tumor to provide a comprehensive overview and minimize the impact of this limitation. In conclusion, our findings underscore the pivotal role of CD147 in modulating the TIME in HCC, highlighting its interplay with OSM and its influence on the Treg landscape. The complex associations we uncovered offer valuable insights into the mechanisms of immune evasion and tumor progression, providing a compelling rationale for further investigation of the CD147-OSM axis as a potential therapeutic target. Future prospective studies encompassing a wide range of molecular and cellular examinations are essential to validate our findings and explore the therapeutic implications. By targeting the CD147-OSM axis and its effect on the TIME, novel therapeutic strategies could be developed to improve outcomes in patients with HCC. Abbreviations ALBI albumin-bilirubin EMT epithelial-mesenchymal transition FFPE formalin-fixed,paraffin-embedded HCC hepatocellular carcinoma MMP matrix metalloproteinase OSM oncostatin M TGF-β transforming growth factor-β TIL tumor-infiltrating lymphocyte TIME tumor-immune microenvironment TMA tissue microarray Treg regulatory T cell. Declarations Funding This work was supported by JSPS KAKENHI (Grant Numbers JP22H02930 and JP23K18246 to KI and Grant Number JP23K14491 to YS), and a research grant from Daiichi Sankyo Co., Ltd. Conflicting interest Yasuyuki Shigematsu, Kengo Takeuchi, and Kentaro Inamura received research grants and materials including the anti-CD147 antibody (clone E1S1V) and a cell array for positive and negative control of CD147 expression from Daiichi Sankyo Co., Ltd., which may represent a conflict of interest. Hiroaki Kanda and Yu Takahashi declare that they have no conflict of interest. Ethics statement All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2008. The study was approved by the Ethics Committee of the Japanese Foundation for Cancer Research (approval reference number: 2017-GA-1136). The requirement for informed consent was waived because of the retrospective study design. Author contributions Yasuyuki Shigematsu: Conceptualization, methodology, investigation, data curation, formal analysis, result interpretation, funding acquisition, project administration, writing–original draft, and writing–review. Hiroaki Kanda: Methodology, investigation, and writing–review. Yu Takahashi: Resources, methodology, and writing–review. Kengo Takeuchi: Supervision, writing–review. Kentaro Inamura: Conceptualization, methodology, investigation, result interpretation, funding acquisition, supervision, writing–original draft, and writing–review. All authors read and approved the final manuscript. Availability of data and material The datasets used and/or analyzed during this study are available from the corresponding authors upon reasonable request. Acknowledgement The authors thank Daiichi Sankyo Co., Ltd. for providing us with anti-CD147 antibody (clone E1S1V) and cell array for positive and negative control of CD147 expression. References Sung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71:209-249. Arnold M, Abnet CC, Neale RE, et al. Global Burden of 5 Major Types of Gastrointestinal Cancer. Gastroenterology. 2020;159:335-349 e315. Ding J, Wen Z. 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Supplementary Files FigS1CD147.tif Fig. S1 Immunohistochemistry of CD8 + , CD4 + , FOXP3 + , and CD20 + cells in hepatocellular carcinoma. Scale bar = 50 μm. FigS2CD147.tif Fig. S2 Correlation between density of oncostatin M (OSM) and tumor-infiltrating lymphocytes (CD8 + , CD4 + , FOXP3 + , and CD20 + cells). Blue dots represent the CD147-low group, whereas red dots represent the CD147-high group. 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-4418866","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":306550874,"identity":"686ee3d1-896e-4807-8941-025b72ac6eae","order_by":0,"name":"Yasuyuki Shigematsu","email":"","orcid":"","institution":"The Cancer Institute of Japanese Foundation for Cancer Research","correspondingAuthor":false,"prefix":"","firstName":"Yasuyuki","middleName":"","lastName":"Shigematsu","suffix":""},{"id":306550875,"identity":"036d9f5c-877d-4deb-a6e2-3bc3de50785a","order_by":1,"name":"Hiroaki Kanda","email":"","orcid":"","institution":"Saitama cancer center","correspondingAuthor":false,"prefix":"","firstName":"Hiroaki","middleName":"","lastName":"Kanda","suffix":""},{"id":306550876,"identity":"7d211a19-6070-477f-b048-a00f235a47fc","order_by":2,"name":"Yu Takahashi","email":"","orcid":"","institution":"The Cancer Institute Hospital of Japanese Foundation for cancer research","correspondingAuthor":false,"prefix":"","firstName":"Yu","middleName":"","lastName":"Takahashi","suffix":""},{"id":306550877,"identity":"70951d4d-996f-4df0-9033-35f83ed59b02","order_by":3,"name":"Kengo Takeuchi","email":"","orcid":"","institution":"The Cancer institute of Japanese Foundation for cancer research","correspondingAuthor":false,"prefix":"","firstName":"Kengo","middleName":"","lastName":"Takeuchi","suffix":""},{"id":306550878,"identity":"f34097fb-7b85-4e34-a77e-dc05fde56a7a","order_by":4,"name":"Kentaro Inamura","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDklEQVRIiWNgGAWjYBACCRiDDUrLMTYfgLAYG3BqgUjBtBgztiUQqQUGEhvYEvA7TLK99/iDn3tsGPikm59u5t1jl97cxvzsA0PFPQbm2ditkeY5l9jY8yyNgU3mmNltnmfJuY1tbMYzGM4UMzDOOYBVi5xEjmEDz4HDDGwSCUAtB5hzG+f3MDOAPMQ4A7sL5eTfGDb+AWtJ/wbUUp/O2MaDX4u0BI9hM8SWHJAthxMIapHsyTGcLXMgjQeopezmnAPHDUF+YUg4k8CDyy8Sx88YfHxzwEZOfkb6thtvDlTLG7YxP2b4UJEgZ4gjxGCAB84yBCkEOonHcAZeHUhAHs6QwKdsFIyCUTAKRhAAALEMWMWPuNp2AAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0001-6444-3861","institution":"The Cancer Institute of Japanese Foundation for Cancer Research","correspondingAuthor":true,"prefix":"","firstName":"Kentaro","middleName":"","lastName":"Inamura","suffix":""}],"badges":[],"createdAt":"2024-05-14 11:36:49","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4418866/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4418866/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":57924183,"identity":"286b07de-910d-4568-bf9b-af9afb0433e8","added_by":"auto","created_at":"2024-06-07 14:02:50","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1231984,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative images of CD147-high and CD147-low hepatocellular carcinoma (HCC). Microscopic images of (A) CD147-high and (B) CD147-low HCC. The upper panel is stained with hematoxylin and eosin, and the lower panel shows immunohistochemistry staining for CD147. In CD147-high HCC, diffuse positivity of CD147 is present circumferentially along the tumor cell membrane. Scale bar = 100 μm.\u003c/p\u003e","description":"","filename":"Fig1CD147.png","url":"https://assets-eu.researchsquare.com/files/rs-4418866/v1/2db541174c24f60cda2abb9f.png"},{"id":57924186,"identity":"7661443e-d302-4ec9-bfdf-9c7fb1cd1dde","added_by":"auto","created_at":"2024-06-07 14:02:50","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":567538,"visible":true,"origin":"","legend":"\u003cp\u003eAssociation between CD147 expression and oncostatin M (OSM)-positive cells in hepatocellular carcinoma (HCC). (A) Representative images of OSM immunohistochemistry in HCC tissues. OSM expression is present in residual macrophages. (B) The box plot shows that the density of OSM-positive cells is higher in CD147-high HCC than in CD147-low HCC (\u003cem\u003eP\u003c/em\u003e = 0.0017).\u003c/p\u003e","description":"","filename":"Fig2CD147.png","url":"https://assets-eu.researchsquare.com/files/rs-4418866/v1/bc4b26ae950013eaeb49117c.png"},{"id":57924790,"identity":"ec27ada0-8d60-426a-8ee4-e644865d268c","added_by":"auto","created_at":"2024-06-07 14:10:50","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":155394,"visible":true,"origin":"","legend":"\u003cp\u003eCorrelation between oncostatin M (OSM)-positive cells and tumor-infiltrating lymphocytes (TILs) in hepatocellular carcinoma (HCC) according to CD147 expression status.\u0026nbsp; (A) Significant associations are present between OSM-positive cells and various TILs in all HCC samples (all \u003cem\u003eP\u003c/em\u003es \u0026lt; 0.0001). \u0026nbsp;(B) These significant associations are also present in CD147-high HCC (all \u003cem\u003eP\u003c/em\u003es \u0026lt; 0.0001). \u0026nbsp;(C) In CD147-low HCC, a significant association is present only for FOXP3\u003csup\u003e+\u003c/sup\u003e cells (\u003cem\u003eP\u003c/em\u003e = 0.0004), whereas CD8\u003csup\u003e+\u003c/sup\u003e cells (\u003cem\u003eP\u003c/em\u003e = 0.58), CD4\u003csup\u003e+\u003c/sup\u003e cells (\u003cem\u003eP\u003c/em\u003e = 0.86), and CD20\u003csup\u003e+\u003c/sup\u003e cells (\u003cem\u003eP\u003c/em\u003e = 0.54) were not significantly associated with OSM-positive cells.\u003c/p\u003e","description":"","filename":"Fig3CD147.png","url":"https://assets-eu.researchsquare.com/files/rs-4418866/v1/d6924ba5e7ae479d9f90d770.png"},{"id":58396007,"identity":"f68a8d53-2615-4749-b00a-2aa3b7ff43fe","added_by":"auto","created_at":"2024-06-15 01:03:19","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2905470,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4418866/v1/4e921e31-1fee-47ed-996c-f4be7b89228a.pdf"},{"id":57924188,"identity":"d5f698f0-4f16-4a36-99ed-ae8cef906378","added_by":"auto","created_at":"2024-06-07 14:02:50","extension":"tif","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":13738401,"visible":true,"origin":"","legend":"\u003cp\u003eFig. S1\u003c/p\u003e\n\u003cp\u003eImmunohistochemistry of CD8\u003csup\u003e+\u003c/sup\u003e, CD4\u003csup\u003e+\u003c/sup\u003e, FOXP3\u003csup\u003e+\u003c/sup\u003e, and CD20\u003csup\u003e+\u003c/sup\u003e cells in hepatocellular carcinoma.\u0026nbsp; Scale bar = 50 μm.\u003c/p\u003e","description":"","filename":"FigS1CD147.tif","url":"https://assets-eu.researchsquare.com/files/rs-4418866/v1/c2856d8d46fdc2788123a073.tif"},{"id":57924184,"identity":"36347352-9dd6-4333-a401-1f5e985c4afc","added_by":"auto","created_at":"2024-06-07 14:02:50","extension":"tif","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":528113,"visible":true,"origin":"","legend":"\u003cp\u003eFig. S2\u003c/p\u003e\n\u003cp\u003eCorrelation between density of oncostatin M (OSM) and tumor-infiltrating lymphocytes (CD8\u003csup\u003e+\u003c/sup\u003e, CD4\u003csup\u003e+\u003c/sup\u003e, FOXP3\u003csup\u003e+\u003c/sup\u003e, and CD20\u003csup\u003e+\u003c/sup\u003e cells).\u0026nbsp; Blue dots represent the CD147-low group, whereas red dots represent the CD147-high group.\u003c/p\u003e","description":"","filename":"FigS2CD147.tif","url":"https://assets-eu.researchsquare.com/files/rs-4418866/v1/212a6b2c163fdee434a56679.tif"}],"financialInterests":"","formattedTitle":"Relationships between CD147 expression, tumor-infiltrating lymphocytes, and oncostatin M in hepatocellular carcinoma","fulltext":[{"header":"Introduction","content":"\u003cp\u003ePrimary liver cancer is the sixth most commonly diagnosed cancer and the third leading cause of cancer-related deaths globally.\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e Hepatocellular carcinoma (HCC), which constitutes 75\u0026ndash;85% of all primary liver cancer cases,\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e generally has a poor prognosis, with a 5-year overall survival rate of approximately 16%. This poor prognosis has remained relatively unchanged over the last three decades,\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e despite advancements in diagnosis and treatment. There is a critical need for a deeper understanding of both tumor cells and the tumor-immune microenvironment (TIME) to develop effective HCC treatment strategies.\u003c/p\u003e \u003cp\u003eRecent studies have highlighted the importance of CD147, a glycoprotein originally known as a regulator of matrix metalloproteinase (MMP) in various tumor types, including HCC.\u003csup\u003e\u003cspan additionalcitationids=\"CR5\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e The overexpression of CD147 in tumor cells has been identified as a predictor of high malignant potential and advanced clinical stage.\u003csup\u003e\u003cspan additionalcitationids=\"CR8\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e CD147 expression in tumor cells induces mesenchymal cells to produce MMPs,\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e which degrade the extracellular matrix and create a niche conducive to tumor invasion and metastasis. Through its interaction with the transforming growth factor-β (TGF-β) signaling pathway, CD147 substantially enhances tumor invasiveness by facilitating the epithelial-mesenchymal transition (EMT) of HCC cells.\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e However, its influence on the TIME remains inadequately explored. Given recent advancements in antitumor immunotherapy, elucidating the association between CD147 expression and the TIME is essential, as it could facilitate the development of effective therapeutic strategies.\u003c/p\u003e \u003cp\u003eOncostatin M (OSM), a pleiotropic inflammatory cytokine produced mainly by macrophages and Kupffer cells in the liver,\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e also promotes EMT and advances HCC progression.\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e The association between CD147 expression and OSM, and their collective effects on the TIME and immune modulation, remains insufficiently explored. This raises questions regarding the potential role of CD147 and OSM in modulating tumor behavior and immune evasion in HCC.\u003c/p\u003e \u003cp\u003eThis study aimed to elucidate the clinicopathological features associated with CD147 expression in HCC and investigate its relationship with the TIME, thereby providing a basis for novel therapeutic strategies for the management of HCC.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePatients and specimens\u003c/h2\u003e \u003cp\u003eSpecimens from surgically resected HCC were obtained from patients at the Cancer Institute Hospital of the Japanese Foundation for Cancer Research in Tokyo, Japan, between January 1, 2005 and December 31, 2016. These patients underwent resection with curative intent for HCC. Patients with congenital and chronic autoimmune liver diseases were excluded. This study included a cohort of 397 patients diagnosed with HCC, selected on the basis of the availability of formalin-fixed, paraffin-embedded (FFPE) tissue specimens for analyses. Clinicopathological variables, including height, weight, sex, preoperative blood test results, and the clinical tumor-node-metastasis (TNM) stage, were extracted from the medical record database. Hepatic functionality was evaluated utilizing the albumin-bilirubin (ALBI) score system.\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e Tumor staging adhered to the 8th edition of the TNM classification by the Unio Internationalis Contra Cancrum.\u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e Approval for the study protocol was obtained from the ethics committee at the Japanese Foundation for Cancer Research (approval number 2017-GA-1136). The requirement for informed consent was waived because of the retrospective study design.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003ePathological evaluation\u003c/h2\u003e \u003cp\u003eHematoxylin and eosin-stained 4 \u0026micro;m sections of FFPE samples from the patients with HCC were reviewed histologically by two experienced liver pathologists (Y.S. and H.K.). The diagnosis of HCC was performed according to the 5th edition of the World Health Organization criteria.\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e Pathological features were evaluated according to the Japanese general rules for clinical and pathological study of primary liver cancer.\u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e A whole section of noncancerous liver tissues without any tumor cells was used to evaluate the background liver tissue of each patient. Hepatic fibrosis was evaluated based on the Brunt criteria.\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eImmunohistochemistry\u003c/h2\u003e \u003cp\u003eTissue microarrays (TMAs) were prepared using FFPE tumor tissues to assess the immunohistochemical features of tumor cells and tumor-infiltrating lymphocytes (TILs), as previously described.\u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e Briefly, designated tissue cores of the donor paraffin blocks were extracted using a coring needle measuring 2 mm in diameter. Subsequently, the tissue cores were arranged in an array on the recipient block using a KIN-1 manual tissue arrayer (Azumaya, Tokyo, Japan). Three tissue cores, representing the primary histology of each tumor, were selected. TMA sections of 4 \u0026micro;m thick were then used for subsequent immunohistochemical evaluations.\u003c/p\u003e \u003cp\u003eImmunohistochemical evaluation of the tumor tissues was performed using anti-CD147 rabbit monoclonal antibody (1:60; clone E1S1V; Cell Signaling Technology, Inc., Danvers, MA, USA) and anti-oncostatin M rabbit polyclonal antibody (1:300; ab198830; Abcam, Cambridge, UK). TILs were assessed using anti-CD8 mouse monoclonal antibody (1:3; clone C8/144B; Nichirei, Tokyo, Japan), anti-CD4 mouse monoclonal antibody (1:2; clone 4B12; Nichirei), anti-CD20 mouse monoclonal antibody (1:800; clone L26; Leica, Wetzlar, Germany), and anti-FOXP3 mouse monoclonal antibody (1:100; clone 236A/E7; Abcam). For positive and negative controls for CD147, we used a cell array obtained from Daiichi Sankyo Co., Ltd., consisting of A431 (Cat #CRL-1555, ATCC, high expression), BxPC-3 (Cat #CRL-1687, ATCC, high expression), HCC1954 (Cat #CRL-2338, ATCC, high expression), ZR-75-1 (Cat #CRL-1500, ATCC, middle expression), MCF-7 (Cat #HTB-22, ATCC, low-middle expression), MDA-MB-453 (Cat #HTB-131, ATCC, low expression), and CHO-K1 cell lines (Cat #CCL-61, ATCC, negative). Cultured cell lines were collected and were fixed with 20% formalin neutral buffer solution (Cat. No. 060-01721, FUJIFILM Wako Pure Chemical Corporation), and were embedded with Histogel (Cat. No. HG-4000-012, Epredia Netherlands B.V.). These blocks were then replaced with paraffin and the slides of thin sections were prepared at Daiichi Sankyo Co., Ltd. Non-cancerous liver tissue was used for positive and negative controls for OSM, CD8, CD4, FOXP3, and CD20. Immunostaining was performed using a Bond-III automated staining system (Leica Microsystems, Buffalo Grove, IL, USA), and antigens were detected using a Bond Polymer Refine Detection Kit (Leica Microsystems).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eEvaluation of protein expression through immunohistochemistry in TMA\u003c/h2\u003e \u003cp\u003eCD147 expression was regarded as positive if circumferential membrane expression was present. CD147 expression in tumor tissues was categorized as high or low, according to the predominant expression pattern observed across the tumor cells, and HCC samples were classified into two groups based on the CD147 expression status: HCC showing high CD147 expression (CD147-high HCC) and HCC showing low CD147 expression (CD147-low HCC). To evaluate the number of OSM-positive cells and TILs, immunostained sections were scanned at \u0026times;40 magnification using a NanoZoomer Digital Pathology System (Hamamatsu Photonics KK, Shizuoka, Japan) at a resolution of 0.55 pixel/\u0026micro;m and semi-automatically digitized using the proprietary NanoZoomer Digital Pathology Image file format. TILs were counted using Fiji, an open-source platform for biological-image analysis.\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e A microscopic field was selected for each tissue core from the scanned TMA images. The mean number of TILs counted in the three tissue cores of each tumor was considered the number of TILs in the tumor.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eStatistical analyses were performed using R version 4.2.0 (R Foundation for Statistical Computing, Vienna, Austria). Continuous variables were reported as means, medians, standard deviations, or standard errors as appropriate. Categorical variables were reported as numbers and percentages. The statistical significance of between-group differences of continuous variables was assessed using either the Mann-Whitney \u003cem\u003eU\u003c/em\u003e test or Student\u0026rsquo;s \u003cem\u003et\u003c/em\u003e-test, and categorical variables were compared using the Fisher\u0026rsquo;s exact test. A two-tailed \u003cem\u003eP\u003c/em\u003e value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003ch2\u003eCD147 expression and clinicopathological features\u003c/h2\u003e \u003cp\u003eThe CD147 staining in the HCC specimens was relatively homogenous (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Of the 397 HCC tissue samples analyzed, 332 (83.6%) showed high CD147 expression. The clinicopathological features are summarized according to CD147 expression status in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. High CD147 expression was associated with advanced clinical stages (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0029). A fibrous capsule or septal fibrosis was more frequently present in CD147-high HCC than in CD147-low HCC (94.0% vs. 86.2%, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.036). No significant differences in CD147 status were observed according to age, sex, history of hepatitis virus infection, tumor differentiation, or presence of background liver cirrhosis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eClinicopathological features of patients with HCC according to CD147 expression status.\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=\"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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eVariable\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTotal (n\u0026thinsp;=\u0026thinsp;397)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eCD147\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLow (n\u0026thinsp;=\u0026thinsp;65)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHigh (n\u0026thinsp;=\u0026thinsp;332)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge (years) \u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.9 (\u0026plusmn;\u0026thinsp;10.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e66.4 (\u0026plusmn;\u0026thinsp;10.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e68.1 (\u0026plusmn;\u0026thinsp;9.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSex \u003csup\u003e\u0026sect;\u003c/sup\u003e\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 \u003cp\u003e0.57\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=\"left\" colname=\"c2\"\u003e \u003cp\u003e90 (22.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17 (26.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e73 (22.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e307 (77.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e48 (73.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e259 (78.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBMI \u003csup\u003e\u0026sect;\u003c/sup\u003e\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 \u003cp\u003e0.24\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e273 (68.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e49 (75.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e224 (67.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25 ≦\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e124 (31.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16 (24.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e108 (32.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHepatitis virus infection \u003csup\u003e\u0026sect;\u003c/sup\u003e\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 \u003cp\u003e0.94\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHBV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e99 (24.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18 (27.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e81 (24.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHCV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e138 (34.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e21 (32.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e117 (35.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHBV\u0026thinsp;+\u0026thinsp;HCV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11 (2.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2 (3.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9 (2.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOther\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e149 (37.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24 (36.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e125 (37.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eALBI \u003csup\u003e\u0026sect;\u003c/sup\u003e\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 \u003cp\u003e0.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGrade 1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e210 (52.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e27 (41.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e183 (55.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGrade 2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e184 (46.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e37 (56.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e147 (44.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGrade 3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3 (0.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1 (1.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2 (0.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNM stage \u003csup\u003e\u0026sect;\u003c/sup\u003e\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 \u003cp\u003e0.0029\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e32 (8.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8 (12.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e24 (7.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e52 (13.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4 (6.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e48 (14.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eII\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e258 (65.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46 (70.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e212 (63.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIIIA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e34 (8.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7 (10.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e27 (8.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIIIB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e21 (5.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0 (0.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21 (6.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTumor differentiation \u003csup\u003e\u0026sect;\u003c/sup\u003e\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 \u003cp\u003e0.59\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWell\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e36 (9.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4 (6.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e32 (9.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eModerately\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e281 (70.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e49 (75.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e232 (69.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePoorly\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e80 (20.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12 (18.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e68 (20.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGrowth pattern \u003csup\u003e\u0026sect;\u003c/sup\u003e\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 \u003cp\u003e0.81\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExpansive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e363 (91.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e59 (90.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e304 (91.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eInfiltrative\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e34 (8.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6 (9.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e28 (8.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFibrous capsule or septal fibrosis in the tumor \u003csup\u003e\u0026sect;\u003c/sup\u003e\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 \u003cp\u003e0.036\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAbsent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29 (7.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9 (13.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20 (6.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePresent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e368 (92.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e56 (86.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e312 (94.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBackground liver cirrhosis \u003csup\u003e\u0026sect;\u003c/sup\u003e\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 \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAbsent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e235 (59.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e39 (60.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e196 (59.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePresent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e162 (40.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26 (40.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e136 (41.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003e\u003csup\u003e\u0026sect;\u003c/sup\u003e Data presented as n (%); \u003csup\u003e\u0026dagger;\u003c/sup\u003e Data presented as mean (\u0026plusmn;\u0026thinsp;standard deviation).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003eAbbreviations: ALBI, albumin-bilirubin; BMI, body mass index; HBV, hepatitis B virus; HCC, hepatocellular carcinoma; HCV, hepatitis C virus; TNM, tumor-node-metastasis.\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\u003eCD147 expression and tumor-infiltrating lymphocytes\u003c/p\u003e \u003cp\u003eTo evaluate the TIME associated with CD147 expression on tumor cells, we quantified densities of CD8\u003csup\u003e+\u003c/sup\u003e, CD4\u003csup\u003e+\u003c/sup\u003e, FOXP3\u003csup\u003e+\u003c/sup\u003e, and CD20\u003csup\u003e+\u003c/sup\u003e cells within the tumor tissues (\u003cb\u003eFig. S1\u003c/b\u003e). Compared with CD147-low HCC, CD147-high HCC contained higher densities of CD4\u003csup\u003e+\u003c/sup\u003e cells (median, 46.0/mm\u003csup\u003e2\u003c/sup\u003e vs. 104.5/mm\u003csup\u003e2\u003c/sup\u003e for CD147-low vs. CD147-high, respectively; \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0012) and FOXP3\u003csup\u003e+\u003c/sup\u003e cells (median, 11.0/mm\u003csup\u003e2\u003c/sup\u003e vs. 17.5/mm\u003csup\u003e2\u003c/sup\u003e for CD147-low HCC vs. CD147-high HCC, respectively; \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0039) in the tumor tissues (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The density of CD8\u003csup\u003e+\u003c/sup\u003e cells and CD20\u003csup\u003e+\u003c/sup\u003e cells did not differ significantly according to the CD147 expression status of the tumor. These observations suggest that CD147 expression on tumor cells may influence TIME through the induction of regulatory T cells (Tregs). Therefore, in HCC, CD147 expression is associated with an immunosuppressive TIME.\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\u003eTumor infiltrating lymphocytes and CD147 in hepatocellular carcinoma.\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=\"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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eType of cell\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eCD147\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLow\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHigh\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCD8\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e127.0 (48.5-178.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e125.5 (34.8-399.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.25\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCD4\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e46.0 (7.8\u0026ndash;114.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e104.5 (22.0-252.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0012\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFOXP3\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.0 (6.0-30.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.5 (8.0-67.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0039\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCD20\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.5 (5.0-79.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.0 (6.0-112.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003eData presented as mean (1st quartile-3rd quartile).\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\u003eCD147 expression on tumor cells and OSM-positive cells\u003c/p\u003e \u003cp\u003eCD147 is implicated in enhancing the invasiveness of hepatocytes by inducing EMT via the TGF-β signaling pathway.\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e Similarly, OSM is recognized for its role in promoting EMT in HCC.\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e To assess the association between CD147 and OSM, we evaluated their expression profiles using immunohistochemistry. OSM was predominantly expressed in stromal cells adjacent to the periphery of the tumor cell cords (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). Immunohistochemical analysis revealed a higher density of OSM-positive cells in CD147-high than CD147-low HCC (median, 61/mm\u003csup\u003e2\u003c/sup\u003e vs. 25/mm\u003csup\u003e2\u003c/sup\u003e; \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0017; Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e2\u003c/span\u003eB), suggesting possible interaction between CD147-expressing tumor cells and OSM-positive cells.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAssociation between OSM-positive cells and FOXP3\u003csup\u003e+\u003c/sup\u003e cells according to CD147 expression\u003c/p\u003e \u003cp\u003eTo assess the effect of OSM on TILs, we investigated the association of OSM-positive cells with CD8\u003csup\u003e+\u003c/sup\u003e, CD4\u003csup\u003e+\u003c/sup\u003e, FOXP3\u003csup\u003e+\u003c/sup\u003e, and CD20\u003csup\u003e+\u003c/sup\u003e cells (\u003cb\u003eFig. S2\u003c/b\u003e). CD8\u003csup\u003e+\u003c/sup\u003e, CD4\u003csup\u003e+\u003c/sup\u003e, FOXP3\u003csup\u003e+\u003c/sup\u003e, and CD20\u003csup\u003e+\u003c/sup\u003e cells were significantly associated with OSM-positive cells, with correlation coefficients of 0.21, 0.34, 0.42, and 0.17, respectively (all \u003cem\u003eP\u003c/em\u003es\u0026thinsp;\u0026lt;\u0026thinsp;0.001; Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). To evaluate the influence of CD147 expression on the associations between OSM and TILs, we stratified the analysis by CD147 expression status. In CD147-high HCC, CD8\u003csup\u003e+\u003c/sup\u003e, CD4\u003csup\u003e+\u003c/sup\u003e, FOXP3\u003csup\u003e+\u003c/sup\u003e, and CD20\u003csup\u003e+\u003c/sup\u003e were significantly associated with OSM-positive cells, with correlation coefficients of 0.22, 0.35, 0.41, and 0.18, respectively, each demonstrating a significant correlation (all \u003cem\u003eP\u003c/em\u003es\u0026thinsp;\u0026lt;\u0026thinsp;0.001; Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). In contrast, in CD147-low HCC, the correlation coefficients for CD8\u003csup\u003e+\u003c/sup\u003e, CD4\u003csup\u003e+\u003c/sup\u003e, FOXP3\u003csup\u003e+\u003c/sup\u003e, and CD20\u003csup\u003e+\u003c/sup\u003e cells were \u0026minus;\u0026thinsp;0.070, 0.023, 0.431, and \u0026minus;\u0026thinsp;0.078, respectively, with only FOXP3\u003csup\u003e+\u003c/sup\u003e cells showing a statistically significant correlation (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0004; Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eC), suggesting that the infiltration of FOXP3\u003csup\u003e+\u003c/sup\u003e cells in tumor tissues may be indirectly mediated through OSM rather than a direct effect of CD147.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study elucidates the potential role of CD147 in HCC, particularly its association with the TIME and its interaction with TILs and OSM. The results demonstrate that high CD147 expression, present in most HCC samples, is significantly associated with advanced disease stages and increased fibrosis, highlighting its potential as a biomarker of aggressive HCC phenotypes. Moreover, the observed increase in FOXP3\u003csup\u003e+\u003c/sup\u003e Tregs in CD147-high HCC tumors suggests a potential role of CD147 in creating an immunosuppressive environment conducive to tumor progression and evasion from immune surveillance. Notably, the observed association between CD147 expression, OSM-positive cells, and FOXP3\u003csup\u003e+\u003c/sup\u003e cells indicates a complex interplay that might contribute to an immunosuppressive TIME, highlighting the importance of the CD147-OSM axis in the modulation of immune responses within the TIME in HCC.\u003c/p\u003e \u003cp\u003eThe association between CD147 expression and TIME in HCC, particularly through the modulation of FOXP3\u003csup\u003e+\u003c/sup\u003e and CD8\u003csup\u003e+\u003c/sup\u003e TILs, illuminates the complex immunological dynamics influenced by CD147. The prognostic significance of these TIL subsets in HCC is well-documented: high CD8\u003csup\u003e+\u003c/sup\u003e TIL counts are associated with more favorable prognosis, signifying effective anti-tumor immunity, whereas high FOXP3\u003csup\u003e+\u003c/sup\u003e TIL counts, indicative of Tregs, are associated with a worse prognosis, reflecting an immunosuppressive milieu that facilitates tumor growth and immune escape.\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e Our findings, indicating pronounced FOXP3\u003csup\u003e+\u003c/sup\u003e cell infiltration in CD147-high HCC tumors, emphasize a skewed balance towards an immunosuppressive state, potentially overriding the antitumor effects of CD8\u003csup\u003e+\u003c/sup\u003e cytotoxic T cells.\u003c/p\u003e \u003cp\u003eThe observed interaction between CD147 and OSM indicates a potential synergistic contribution to EMT, thereby influencing tumor invasiveness and metastasis. CD147 is known to facilitate EMT via the TGF-β signaling pathway,\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e whereas OSM activates STAT3, which also triggers EMT in cancer cells.\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e OSM-activated STAT3 cooperates with TGF-β to induce mesenchymal stem cell properties in breast cancer and pancreatic cancer.\u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e Although direct evidence of CD147 and OSM interaction in HCC has not been established, the correlation between high CD147 expression and increased OSM-positive cell density in the TIME suggest a cooperative interaction that may promote EMT. This relationship highlights a complex interplay of factors determining HCC progression and suggests novel potential therapeutic targets.\u003c/p\u003e \u003cp\u003eThe observed positive correlation between OSM-positive cells and FOXP3\u003csup\u003e+\u003c/sup\u003e cells suggests that OSM may promote immunosuppression by Treg induction. This relationship highlights the complex relationship between cytokine signals and the regulation of the immune system within the TIME. Given the strong association between the density of OSM-positive cells in TIME and CD147 expression in tumor cells, the increased infiltration of FOXP3\u003csup\u003e+\u003c/sup\u003e cells in CD147-high HCC may be influenced by OSM. Although the exact mechanism by which OSM increases the number of FOXP3\u003csup\u003e+\u003c/sup\u003e Tregs remains elusive, existing knowledge suggests that OSM may indirectly contribute to the expansion of Tregs by inhibiting IL-17/Th17 activity in CD4\u003csup\u003e+\u003c/sup\u003e T cells.\u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eThis study has several limitations. First, the retrospective nature of this study and restricting the study population to patients undergoing hepatectomy may have introduced inherent bias. Consecutive patients were included to mitigate sampling bias. Second, although the TMA method facilitated the immunohistochemical assessment of both tumor cells and the TIME, it might not have fully captured the heterogeneity of the entire tumor. To make the results more representative of the tumors, we carefully selected three tissue cores per tumor to provide a comprehensive overview and minimize the impact of this limitation.\u003c/p\u003e \u003cp\u003eIn conclusion, our findings underscore the pivotal role of CD147 in modulating the TIME in HCC, highlighting its interplay with OSM and its influence on the Treg landscape. The complex associations we uncovered offer valuable insights into the mechanisms of immune evasion and tumor progression, providing a compelling rationale for further investigation of the CD147-OSM axis as a potential therapeutic target. Future prospective studies encompassing a wide range of molecular and cellular examinations are essential to validate our findings and explore the therapeutic implications. By targeting the CD147-OSM axis and its effect on the TIME, novel therapeutic strategies could be developed to improve outcomes in patients with HCC.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eALBI\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ealbumin-bilirubin\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eEMT\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eepithelial-mesenchymal transition\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eFFPE\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eformalin-fixed,paraffin-embedded\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eHCC\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ehepatocellular carcinoma\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eMMP\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ematrix metalloproteinase\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eOSM\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eoncostatin M\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eTGF-β\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003etransforming growth factor-β\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eTIL\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003etumor-infiltrating lymphocyte\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eTIME\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003etumor-immune microenvironment\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eTMA\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003etissue microarray\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cb\u003eTreg\u003c/b\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eregulatory T cell.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003eFunding\u003c/p\u003e\n\u003cp\u003eThis work was supported by JSPS KAKENHI (Grant Numbers JP22H02930 and JP23K18246 to KI and Grant Number JP23K14491 to YS), and a research grant from Daiichi Sankyo Co., Ltd.\u003c/p\u003e\n\u003cp\u003eConflicting interest\u003c/p\u003e\n\u003cp\u003eYasuyuki Shigematsu, Kengo Takeuchi, and Kentaro Inamura received research grants and materials including the anti-CD147 antibody (clone E1S1V) and a cell array for positive and negative control of CD147 expression from Daiichi Sankyo Co., Ltd., which may represent a conflict of interest. \u0026nbsp;Hiroaki Kanda and Yu Takahashi declare that they have no conflict of interest.\u003c/p\u003e\n\u003cp\u003eEthics statement\u003c/p\u003e\n\u003cp\u003eAll procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2008. \u0026nbsp;The study was approved by the Ethics Committee of the Japanese Foundation for Cancer Research (approval reference number: 2017-GA-1136). \u0026nbsp;The requirement for informed consent was waived because of the retrospective study design.\u003c/p\u003e\n\u003cp\u003eAuthor contributions\u003c/p\u003e\n\u003cp\u003eYasuyuki Shigematsu: Conceptualization, methodology, investigation, data curation, formal analysis, result interpretation, funding acquisition, project administration, writing–original draft, and writing–review. \u0026nbsp; \u0026nbsp;Hiroaki Kanda: Methodology, investigation, and writing–review. \u0026nbsp;Yu Takahashi: Resources, methodology, and writing–review. \u0026nbsp;Kengo Takeuchi: Supervision, writing–review. \u0026nbsp;Kentaro Inamura: Conceptualization, methodology, investigation, result interpretation, funding acquisition, supervision, writing–original draft, and writing–review. \u0026nbsp;All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003eAvailability of data and material\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed during this study are available from the corresponding authors upon reasonable request.\u003c/p\u003e\n\u003cp\u003eAcknowledgement\u003c/p\u003e\n\u003cp\u003eThe authors thank Daiichi Sankyo Co., Ltd. for providing us with anti-CD147 antibody (clone E1S1V) and cell array for positive and negative control of CD147 expression.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. \u003cem\u003eCA Cancer J Clin. \u003c/em\u003e2021;71:209-249.\u003c/li\u003e\n\u003cli\u003eArnold M, Abnet CC, Neale RE, et al. Global Burden of 5 Major Types of Gastrointestinal Cancer. \u003cem\u003eGastroenterology. \u003c/em\u003e2020;159:335-349 e315.\u003c/li\u003e\n\u003cli\u003eDing J, Wen Z. Survival improvement and prognosis for hepatocellular carcinoma: analysis of the SEER database. \u003cem\u003eBMC Cancer. \u003c/em\u003e2021;21:1157.\u003c/li\u003e\n\u003cli\u003eLandras A, Reger de Moura C, Jouenne F, et al. CD147 Is a Promising Target of Tumor Progression and a Prognostic Biomarker. \u003cem\u003eCancers (Basel). \u003c/em\u003e2019;11:1803.\u003c/li\u003e\n\u003cli\u003eHuang D, Rao D, Jin Q, et al. Role of CD147 in the development and diagnosis of hepatocellular carcinoma. \u003cem\u003eFront Immunol. \u003c/em\u003e2023;14:1149931.\u003c/li\u003e\n\u003cli\u003eNyalali AMK, Leonard AU, Xu Y, et al. CD147: an integral and potential molecule to abrogate hallmarks of cancer. \u003cem\u003eFront Oncol. \u003c/em\u003e2023;13:1238051.\u003c/li\u003e\n\u003cli\u003ePeng F, Li H, You Q, et al. CD147 as a Novel Prognostic Biomarker for Hepatocellular Carcinoma: A Meta-Analysis. \u003cem\u003eBiomed Res Int. \u003c/em\u003e2017;2017:5019367.\u003c/li\u003e\n\u003cli\u003eZhang Q, Zhou J, Ku XM, et al. Expression of CD147 as a significantly unfavorable prognostic factor in hepatocellular carcinoma. \u003cem\u003eEur J Cancer Prev. \u003c/em\u003e2007;16:196-202.\u003c/li\u003e\n\u003cli\u003eZhu S, Li Y, Zhang Y, et al. Expression and clinical implications of HAb18G/CD147 in hepatocellular carcinoma. \u003cem\u003eHepatol Res. \u003c/em\u003e2015;45:97-106.\u003c/li\u003e\n\u003cli\u003eBiswas C, Zhang Y, DeCastro R, et al. The human tumor cell-derived collagenase stimulatory factor (renamed EMMPRIN) is a member of the immunoglobulin superfamily. \u003cem\u003eCancer Res. \u003c/em\u003e1995;55:434-439.\u003c/li\u003e\n\u003cli\u003eXu J, Xu HY, Zhang Q, et al. HAb18G/CD147 functions in invasion and metastasis of hepatocellular carcinoma. \u003cem\u003eMol Cancer Res. \u003c/em\u003e2007;5:605-614.\u003c/li\u003e\n\u003cli\u003eRu NY, Wu J, Chen ZN, et al. HAb18G/CD147 is involved in TGF-beta-induced epithelial-mesenchymal transition and hepatocellular carcinoma invasion. \u003cem\u003eCell Biol Int. \u003c/em\u003e2015;39:44-51.\u003c/li\u003e\n\u003cli\u003eLevy MT, Trojanowska M, Reuben A. Oncostatin M: a cytokine upregulated in human cirrhosis, increases collagen production by human hepatic stellate cells. \u003cem\u003eJ Hepatol. \u003c/em\u003e2000;32:218-226.\u003c/li\u003e\n\u003cli\u003eZnoyko I, Sohara N, Spicer SS, et al. Expression of oncostatin M and its receptors in normal and cirrhotic human liver. \u003cem\u003eJ Hepatol. \u003c/em\u003e2005;43:893-900.\u003c/li\u003e\n\u003cli\u003eDi Maira G, Foglia B, Napione L, et al. Oncostatin M is overexpressed in NASH-related hepatocellular carcinoma and promotes cancer cell invasiveness and angiogenesis. \u003cem\u003eJ Pathol. \u003c/em\u003e2022;257:82-95.\u003c/li\u003e\n\u003cli\u003eShigematsu Y, Tanaka K, Amori G, et al. Potential involvement of oncostatin M in the immunosuppressive tumor-immune microenvironment in hepatocellular carcinoma with vessels encapsulating tumor clusters. \u003cem\u003eHepatol Res. \u003c/em\u003e2023.\u003c/li\u003e\n\u003cli\u003eChan AW, Kumada T, Toyoda H, et al. Integration of albumin-bilirubin (ALBI) score into Barcelona Clinic Liver Cancer (BCLC) system for hepatocellular carcinoma. \u003cem\u003eJ Gastroenterol Hepatol. \u003c/em\u003e2016;31:1300-1306.\u003c/li\u003e\n\u003cli\u003eAmin MB, Greene FL, Edge SB, et al. The Eighth Edition AJCC Cancer Staging Manual: Continuing to build a bridge from a population-based to a more \u0026quot;personalized\u0026quot; approach to cancer staging. \u003cem\u003eCA Cancer J Clin. \u003c/em\u003e2017;67:93-99.\u003c/li\u003e\n\u003cli\u003eTorbenson M, Ng IOL, Park YN, Roncalli M, Sakamoto M. \u003cem\u003eHepatocellular carcinoma. In: WHO Classification of Tumours Editorial Board, editor. Digestive system tumours.\u003c/em\u003e: Lyon: International Agency for Research on Cancer; 2019.\u003c/li\u003e\n\u003cli\u003eKudo M, Kawamura Y, Hasegawa K, et al. Management of Hepatocellular Carcinoma in Japan: JSH Consensus Statements and Recommendations 2021 Update. \u003cem\u003eLiver Cancer. \u003c/em\u003e2021;10:181-223.\u003c/li\u003e\n\u003cli\u003eBrunt EM, Tiniakos DG. Histopathology of nonalcoholic fatty liver disease. \u003cem\u003eWorld J Gastroenterol. \u003c/em\u003e2010;16:5286-5296.\u003c/li\u003e\n\u003cli\u003eShigematsu Y, Amori G, Kanda H, et al. Decreased ARG1 expression as an adverse prognostic phenotype in non-alcoholic non-virus-related hepatocellular carcinoma. \u003cem\u003eVirchows Arch. \u003c/em\u003e2022;481:253-263.\u003c/li\u003e\n\u003cli\u003eShigematsu Y, Inamura K, Yamamoto N, et al. Impact of CDX2 expression status on the survival of patients after curative resection for colorectal cancer liver metastasis. \u003cem\u003eBMC Cancer. \u003c/em\u003e2018;18:980.\u003c/li\u003e\n\u003cli\u003eSchindelin J, Arganda-Carreras I, Frise E, et al. Fiji: an open-source platform for biological-image analysis. \u003cem\u003eNat Methods. \u003c/em\u003e2012;9:676-682.\u003c/li\u003e\n\u003cli\u003eCaligiuri A, Gitto S, Lori G, et al. Oncostatin M: From Intracellular Signaling to Therapeutic Targets in Liver Cancer. \u003cem\u003eCancers (Basel). \u003c/em\u003e2022;14.[page 4211]\u003c/li\u003e\n\u003cli\u003eYao W, He JC, Yang Y, et al. The Prognostic Value of Tumor-infiltrating Lymphocytes in Hepatocellular Carcinoma: a Systematic Review and Meta-analysis. \u003cem\u003eSci Rep. \u003c/em\u003e2017;7:7525.\u003c/li\u003e\n\u003cli\u003eGiannelli G, Koudelkova P, Dituri F, et al. Role of epithelial to mesenchymal transition in hepatocellular carcinoma. \u003cem\u003eJ Hepatol. \u003c/em\u003e2016;65:798-808.\u003c/li\u003e\n\u003cli\u003eJunk DJ, Bryson BL, Smigiel JM, et al. Oncostatin M promotes cancer cell plasticity through cooperative STAT3-SMAD3 signaling. \u003cem\u003eOncogene. \u003c/em\u003e2017;36:4001-4013.\u003c/li\u003e\n\u003cli\u003eSmigiel JM, Parameswaran N, Jackson MW. Potent EMT and CSC Phenotypes Are Induced By Oncostatin-M in Pancreatic Cancer. \u003cem\u003eMol Cancer Res. \u003c/em\u003e2017;15:478-488.\u003c/li\u003e\n\u003cli\u003eSon HJ, Lee SH, Lee SY, et al. Oncostatin M Suppresses Activation of IL-17/Th17 via SOCS3 Regulation in CD4+ T Cells. \u003cem\u003eJ Immunol. \u003c/em\u003e2017;198:1484-1491.\u003c/li\u003e\n\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":"CD20, CD4, CD8, FOXP3, hepatic fibrosis, immunity, immunohistochemistry, liver cancer, regulatory T cells, tumor-immune microenvironment","lastPublishedDoi":"10.21203/rs.3.rs-4418866/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4418866/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003ePurpose\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn hepatocellular carcinoma (HCC), CD147 expression contributes to tumor malignancy; however, its relationship with the tumor-immune microenvironment (TIME) remains unclear. This study aimed to elucidate the clinicopathological characteristics associated with CD147 expression in HCC and investigate its association with the TIME, specifically its association with tumor-infiltrating lymphocytes (TILs) and oncostatin M (OSM).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUsing 397 HCC specimens from patients undergoing curative-intent resection, we assessed CD147 expression in tumor cells and quantified OSM-positive cells and various TILs (CD8\u003csup\u003e+\u003c/sup\u003e, CD4\u003csup\u003e+\u003c/sup\u003e, FOXP3\u003csup\u003e+\u003c/sup\u003e, and CD20\u003csup\u003e+\u003c/sup\u003e cells) in the TIME. Using tissue microarrays, these assessments were performed through immunohistochemical analysis. We investigated the associations between CD147 expression status, the density of OSM-positive cells, and the densities of various TILs.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHigh CD147 expression, found in 332 specimens (83.6%), was associated with advanced clinical stage (\u003cem\u003eP\u003c/em\u003e = 0.0029), fibrosis (\u003cem\u003eP\u003c/em\u003e = 0.036), and higher densities of FOXP3\u003csup\u003e+\u003c/sup\u003e cells (\u003cem\u003eP\u003c/em\u003e = 0.0039), CD4\u003csup\u003e+\u003c/sup\u003e cells (\u003cem\u003eP\u003c/em\u003e = 0.0012), and OSM-positive cells (\u003cem\u003eP\u003c/em\u003e = 0.0017). In CD147-high tumors, OSM-positive cell density was associated with all assessed TIL subsets (CD8\u003csup\u003e+\u003c/sup\u003e, CD4\u003csup\u003e+\u003c/sup\u003e, FOXP3\u003csup\u003e+\u003c/sup\u003e, and CD20\u003csup\u003e+\u003c/sup\u003e cells; all \u003cem\u003eP\u003c/em\u003es \u0026lt; 0.001), whereas in CD147-low tumors, OSM-positive cell density was associated only with FOXP3\u003csup\u003e+\u003c/sup\u003e cells (\u003cem\u003eP\u003c/em\u003e = 0.0004).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn HCC, CD147 expression is associated with an immunosuppressive TIME, characterized by increased FOXP3\u003csup\u003e+\u003c/sup\u003e regulatory T cells and an association with OSM-positive cells. These results elucidate the potential mechanisms through which CD147 facilitates tumor immune evasion, suggesting the CD147-OSM axis as a promising target for therapeutic intervention in HCC.\u003c/p\u003e","manuscriptTitle":"Relationships between CD147 expression, tumor-infiltrating lymphocytes, and oncostatin M in hepatocellular carcinoma","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-07 14:02:45","doi":"10.21203/rs.3.rs-4418866/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":"5d9d8d07-3f63-4e56-aae4-94cea4b47d06","owner":[],"postedDate":"June 7th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-06-15T00:55:11+00:00","versionOfRecord":[],"versionCreatedAt":"2024-06-07 14:02:45","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4418866","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4418866","identity":"rs-4418866","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}