FZD7通过β-Catenin/CCNA2信号轴促进肝细胞癌的增殖

FZD7-Mediated Promotion of the Proliferation of Hepatocellular Carcinoma through the β-Catenin/CCNA2 Signaling Axis

摘要: 本研究主要探讨卷曲蛋白7 (FZD7)在肝癌中的表达变化及其对人肝细胞癌细胞增殖的影响。利用公共数据库分析肝癌组织与癌旁组织中FZD7表达差异。构建小鼠肝癌模型，通过IHC染色分析肝癌组织中FZD7蛋白表达。利用基因克隆和载体构建获得重组FZD7过表达和干扰慢病毒载体，并通过慢病毒包装、感染和药物筛选获得FZD7过表达和敲低肝癌细胞株。利用实时无标记细胞分析系统检测肝细胞癌细胞的增殖能力，并用WB检测细胞内源性CCNA2表达变化。生物信息学分析结果及肝癌模型证实肝癌组织中FZD7显著高表达，细胞水平上过表达或敲低FZD7，能有效促进或抑制癌细胞的生长增殖。癌细胞中过表达FZD7能提高β-Catenin核移位并促进下游靶基因CCNA2表达。肝细胞癌细胞中高表达FZD7会促进细胞生长增殖，使其有望成为肝细胞癌诊断与治疗的潜在靶点。

Abstract: The aim of this study is to investigate the dysregulation of Frizzled-7 (FZD7) in hepatocellular carcinoma (HCC) and its functional role on tumor cell proliferation. We first analyzed FZD7 expression profiles in HCC versus adjacent non-tumorous liver tissues using public databases. Next, we established a murine orthotopic HCC model and validated FZD7 protein expression levels in tumor tissues via immunohistochemistry (IHC). To assess causality, we generated lentiviral vectors for stable FZD7 overexpression and shRNA-mediated knockdown through standardized molecular cloning and viral packaging protocols; isogenic HCC cell lines with consistent genetic backgrounds were established following puromycin selection. Cell proliferation dynamics were quantified in real time using a label-free, impedance-based cellular analysis system (xCELLigence), while endogenous CCNA2 protein levels were examined by western blot. Bioinformatic analyses and in vivo experimental data collectively demonstrate significant upregulation of FZD7 in HCC tissues. Functionally, FZD7 overexpression enhances, whereas its knockdown suppresses HCC cell proliferation. Mechanistically, elevated FZD7 promotes β-Catenin nuclear translocation and subsequent transcriptional activation of the downstream target gene CCNA2. High expression of FZD7 in hepatocellular carcinoma cells promotes tumor cell growth and proliferation, suggesting its potential as a diagnostic biomarker and therapeutic target for hepatocellular carcinoma.