引用文本：李智超, 柯立鑫, 林俊龙, 等. 甘氨脱氧胆酸通过法尼醇X受体/上皮-间质转化信号轴抑制肝细胞癌进展[J/CD]. 消化肿瘤杂志（电子版）, 2026, 18(1): 131-144.

作者：李智超，柯立鑫，林俊龙，高凯，华赟鹏

单位：中山大学附属第一医院肝胆胰外科中心，广东 广州 510080

Authors：Li Zhichao, Ke Lixin, Lin Junlong, Gao Kai, Hua Yunpeng

Unit：Center of Hepato-Pancreato-Biliary Surgery, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, Guangdong, China

摘要：

目的 探讨甘氨脱氧胆酸（glycodeoxycholic acid，GDCA）对肝癌细胞增殖、侵袭和转移的影响及其可能的分子机制。方法 构建裸鼠肝细胞癌皮下种植瘤模型和肺转移模型，观察GDCA治疗[200 mg/(kg·d)]对肝细胞癌生长、转移的影响，并通过免疫组织化学染色分析种植瘤中法尼醇X受体（farnesoid X receptor，FXR）的表达。利用Swiss Target Prediction数据库预测GDCA潜在的作用靶点，应用AutoDockTools进行分子对接和结合能计算。使用Kaplan-Meier plotter数据库分析FXR表达对肝细胞癌患者总生存期和无进展生存期的影响。蛋白质印迹法检测经GDCA（0.5 mmol/L）处理后肝癌细胞系（SNU-449、HUH-7、LM3、97H）中FXR的表达。通过shRNA技术建立敲低FXR的LM3肝癌细胞株，采用克隆集落形成实验、CCK-8细胞增殖实验、Transwell细胞迁移实验、细胞划痕实验检测肝癌细胞的增殖、转移能力，并用蛋白质印迹法检测敲低FXR后LM3细胞的上皮-间质转化（epithelial-mesenchymal transition，EMT）相关标志物的表达情况。结果 GDCA能有效抑制肝细胞癌皮下种植瘤和肺转移灶的生长。Swiss Target Prediction数据库预测GDCA最可能的结合靶点为FXR，分子对接显示两者具有高结合活性（结合能为-8.2 kcal/mol）。Kaplan-Meier plotter数据库中FXR高表达的肝细胞癌患者较低表达患者的总生存期（HR=0.52，95%CI 0.33~0.82，P=0.004 5）和无进展生存期（HR=0.63，95%CI 0.47~0.85，P=0.002 2）更长。在GDCA给药治疗后的裸鼠皮下瘤组织中，以及经GDCA处理的肝癌细胞系中FXR表达均上调。此外，在FXR敲低后，LM3细胞的增殖、转移能力增强，FXR的敲低逆转了GDCA对肝癌细胞增殖和转移的抑制作用，且呈现较高的间质细胞标志物波形蛋白表达。结论 GDCA通过FXR减缓EMT的进程，从而抑制肝细胞癌的进展，GDCA治疗有望成为肝细胞癌的一种辅助治疗方法。

关键词：肝细胞癌；胆汁酸；法尼醇X受体；上皮-间质转化

Abstract：

Objective To investigate the antitumor effects of glycodeoxycholic acid (GDCA) on hepatocellular carcinoma (HCC) and to delineate the underlying molecular mechanisms. Method Subcutaneous xenograft and experimental lung metastasis models of HCC were established in nude mice to evaluate the in vivo efficacy of GDCA [200 mg/(kg·d)]. Tumor burden and metastatic spread were monitored longitudinally, and the farnesoid X receptor (FXR) expression in resected tumors was quantified by immunohistochemistry. Putative GDCA targets were predicted using the Swiss Target Prediction platform, and binding affinity to FXR was computed by molecular docking (AutoDockTools). Prognostic significance of FXR transcriptional levels was analyzed in the Kaplan-Meier Plotter HCC cohort. In vitro, FXR protein expression was determined by western blotting in four human HCC cell lines (SNU-449, HUH-7, LM3, 97H) after GDCA (0.5 mmol/L) exposure. Stable FXR knockdown LM3 cells were generated via shRNA transduction; clonogenic, CCK-8, Transwell and wound-healing assays were employed to assess proliferative and invasive capacities. Epithelial–mesenchymal transition (EMT) status was evaluated by western blotting for E-cadherin, N-cadherin and vimentin. Result GDCA administration markedly suppressed both subcutaneous tumor growth and pulmonary metastatic colonization. In silico analysis using the Swiss Target Prediction database identified FXR as the top-ranking high-confidence target of GDCA, exhibiting robust binding energy (∆G=−8.2 kcal/ mol). High intra-tumoral FXR mRNA levels were associated with prolonged overall survival (HR=0.52, 95%CI 0.33-0.82, P=0.004 5) and progression-free survival (HR=0.63, 95%CI 0.47-0.85, P=0.002 2) in patients with HCC. GDCA treatment up-regulated FXR expression in xenograft tissues and in all tested HCC cell lines. Genetic ablation of FXR reversed GDCA-mediated suppression of proliferation and motility, concomitant with acquisition of a mesenchymal phenotype highlighted by increased vimentin expression. Conclusion GDCA impedes HCC progression by activating FXR-dependent signaling, thereby retarding the EMT process. These findings provide a mechanistic rationale for repositioning GDCA as an adjunctive therapeutic agent in HCC. 

Key words：Hepatocellular carcinoma; Bile acid; Farnesoid X receptor; Epithelial-mesenchymal transition

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