Zhang Yang, Zhang Yan, Shi Xiao-Jun, Li Jun-Xiang, Wang Lin-Heng, Xie Chun-E, Wang Yun-Liang
Department of Gastroenterology, Dong Fang Hospital, Beijing University of Chinese Medicine, Beijing, China.
Graduate School, Beijing University of Chinese Medicine, Beijing, China.
Front Oncol. 2022 Feb 17;12:836333. doi: 10.3389/fonc.2022.836333. eCollection 2022.
Hepatocellular carcinoma (HCC) is a highly invasive disease with a high mortality rate. Our previous study found that Chenodeoxycholic acid CDCA) as an endogenous metabolite can enhance the anti-tumor effect. Sorafenib has limited overall efficacy as a first-line agent in HCC, and combined with CDCA may improve its efficacy.
HepG2 cells and Balb/c nude mice were used respectively for and experiments. Flow cytometry, Western blotting, HE and immunohistochemical staining and immunofluorescence were used to study the effects of CDCA combined with sorafenib on HepG2 cell growth and apoptosis-related proteins. Magnetic bead coupling, protein profiling and magnetic bead immunoprecipitation were used to find the targets of CDCA action. The effect of CDCA on EGFR/Stat3 signaling pathway was further verified by knocking down Stat3 and EGFR. Finally, fluorescence confocal, and molecular docking were used to study the binding site of CDCA to EGFR.
In this study, we found that CDCA enhanced the effect of sorafenib in inhibiting the proliferation, migration and invasion of HepG2 cells. Magnetic bead immunoprecipitation and protein profiling revealed that CDCA may enhance the effect of sorafenib by affecting the EGFR/Stat3 signaling pathway. Further results from and gene knockdown experiments, confocal experiments and molecular docking showed that CDCA enhances the efficacy of sorafenib by binding to the extracellular structural domain of EGFR.
This study reveals the mechanism that CDCA enhances the inhibitory effect of sorafenib on HepG2 cell growth and , providing a potential new combination strategy for the treatment of HCC.
肝细胞癌（HCC）是一种具有高侵袭性和高死亡率的疾病。我们之前的研究发现，鹅去氧胆酸（CDCA）作为一种内源性代谢产物可增强抗肿瘤作用。索拉非尼作为HCC一线治疗药物的总体疗效有限，与CDCA联合使用可能会提高其疗效。
分别使用HepG2细胞和Balb/c裸鼠进行体外和体内实验。采用流式细胞术、蛋白质印迹法、苏木精-伊红（HE）染色、免疫组织化学染色和免疫荧光法研究CDCA联合索拉非尼对HepG2细胞生长及凋亡相关蛋白的影响。运用磁珠偶联、蛋白质谱分析和磁珠免疫沉淀法寻找CDCA的作用靶点。通过敲低Stat3和EGFR进一步验证CDCA对EGFR/Stat3信号通路的影响。最后，采用荧光共聚焦和分子对接技术研究CDCA与EGFR的结合位点。
在本研究中，我们发现CDCA增强了索拉非尼对HepG2细胞增殖、迁移和侵袭的抑制作用。磁珠免疫沉淀和蛋白质谱分析表明，CDCA可能通过影响EGFR/Stat3信号通路增强索拉非尼的作用。Stat3和EGFR基因敲低实验、共聚焦实验及分子对接的进一步结果显示，CDCA通过与EGFR的细胞外结构域结合增强索拉非尼的疗效。
本研究揭示了CDCA增强索拉非尼对HepG2细胞生长抑制作用的机制，为HCC治疗提供了一种潜在的新联合策略。
