基于TGF-β/Artemin信号轴探讨健脾解毒方协同索拉菲尼调控EMT抑制肝癌侵袭转移的效应及机制

Hepatocellular carcinoma (HCC), with a high incidence and poor prognosis in China, is characterized by vulnerable invasion and metastasis. Most patients have reached the advanced stage when they are diagnosed. Sorafenib is one of the few targeted therapies for advanced hepatocellular carcinoma in clinic at present, but the existing clinical data show that it has toxic side effects such as liver damage and drug resistance. Jianpi Jiedu Prescription is an effective prescription for clinical treatment of HCC. Our previous study has found that Jianpi Jiedu Prescription can inhibit the proliferation of HCC cells and prolong the survival time of the model with tumors. Pre-experiments indicated that Jianpi Jiedu Prescription combined with sorafenib could effectively inhibit the growth of HCC cells. However, the efficacy of drug combination and its regulatory role in TGF-β/Artemin and EMT need to be further elucidated. This study aims to: 1) evaluate the efficacy of drug combination, its effects in inhibiting the invasion and metastasis of HCC, and its correlation with the regulation of the TGF-β/Artemin signal axis and EMT through in vivo and in vitro experiments, 2) analyze the main active ingredients of Jianpi Jiedu Prescription and the similarities and differences of the sites of interaction between sorafenib and target protein by molecular docking technique. This study can provide an experimental basis for the clinical application of Jianpi Jiedu Prescription combined with antineoplastic drugs in the treatment of HCC, and also provide reference for the study on the synergistic mechanism of other prescriptions and antineoplastic drugs.

肝癌发病率高、早期诊断率低和易侵袭转移，大多数患者确诊时已达晚期。索拉非尼是目前临床上为数不多的针对晚期肝癌的靶向治疗药物之一，但现有临床数据显示其存在肝损害等毒副作用和耐药情况。健脾解毒方是临床治疗肝癌的有效方剂；前期研究证实健脾解毒方具有抑制肝癌细胞增殖、延长带瘤生存时间和增强机体免疫力等作用。为了构建和评价中西药联合应用抗肝癌方案，通过预实验初步证实健脾解毒方联合索拉非尼能有效抑制肝癌生长，但联合用药效应和对TGF-β/Artemin信号轴、EMT的调控作用有待进一步阐明。本项目拟：①通过体内、体外实验评价联合用药效应，及其抗肝癌作用与调控TGF-β/Artemin信号轴逆转EMT的相关性；②应用分子对接技术，分析健脾解毒方主要活性成分、索拉非尼与靶标蛋白发生相互作用的位点异同。为临床应用健脾解毒方联合抗肿瘤药物治疗肝癌提供实验依据；亦可为中医药联合西药的协同增效机理研究提供借鉴。

目前临床对于中晚期肝癌的疗效尚有待进一步提高，单用西药或中药的效果并不理想。索拉非尼因其存在耐药与肝损害等毒副作用限制了其临床有效性。健脾解毒方是临床治疗肝癌有效的方剂，但其作用机制尚未完全明确。肝癌具有侵袭性强、易转移的特点，本项目以TGF-β/Artemin、PI3K/Akt信号通路与EMT相关蛋白为研究切入点，采用网络药理学、分子对接与实验验证手段，探索健脾解毒方与索拉非尼抑制肝癌侵袭转移的作用与机制。本项目发现：健脾解毒方与索拉非尼可以抑制肝癌细胞的增殖、侵袭转移，促进肝癌细胞凋亡；健脾解毒方抑制肝癌细胞侵袭转移的机制，可能与抑制TGF-β/Artemin信号轴及其下游PI3K/Akt信号通路的激活，进而阻断肝癌细胞EMT进程相关。分子对接分析显示：健脾解毒方活性成分对TGF-β/Artemin信号轴的主要因子TGF-β1、PI3K与Akt的作用，与索拉非尼既有相同作用位点、也存在不同的作用位点。健脾解毒方与索拉非尼是值得深入探索的治疗肝癌联合用药方案之一。