蟾皮通过DCP-c-Met通路调控EMT抑制肝癌侵袭转移的分子机制研究

Epithelial-mesenchymal transition (EMT) is the foundation for invasion and metastasis of cancer cells. Preventing EMT is very important for controlling invasion and metastasis of cancer. c-Met is one of the important targets for promoting invasion and metastasis of hepatocarcinoma. Previous studies have shown that DCP is a key factor for stimulating c-Met signaling pathway and Cinobufacini has the effect of inhibiting DCP-induced proliferation of hepatocarcinoma cells. Moreover, Both EMT and invasion and metastasis were inhibited by Cinobufacini. However, its anticancer mechanism is still unclear. Thus, in the current study, we will investigate whether Cinobufacini has the effect of inhibiting EMT in HCC by regulating DCP-c-Met signaling pathway. Firstly, knockdown of c-Met gene by siRNA in HCC cell lines will be established and DCP will be used to stimulating EMT. Then the expressions of EMT biomarker, ERK1/2 MAPK pathway related molecules, and MMPs will be detected. Secondly, the intervention effect of Cinobufacini on EMT of HCC cells via DCP/c-MET signaling pathway will be conducted in vitro and in vivo. Taken together, the current study will contribute to illuminate the regulation effect of DCP/c-Met signaling pathway on EMT in HCC cells and the intervention effect of Cinobufacini, which will be benefit for understanding the pathogenesis of HCC and developing anti-HCC drugs.

上皮细胞-间充质转化（EMT）是肿瘤侵袭转移的关键途径，c-Met是抗肿瘤侵袭转移的重要靶点，前期发现脱-γ-羧基凝血酶原（DCP）是激活c-Met信号通路的关键因素，中药蟾皮能抑制DCP诱导的肝癌细胞增殖，还能抑制肝癌细胞EMT及侵袭转移，然具体分子机制尚不明确。本课题旨在前期基础上探讨蟾皮通过DCP-c-Met通路调控EMT抑制肝癌侵袭转移的分子机制。首先siRNA干扰肝癌细胞c-Met表达并给予DCP诱导，通过检测EMT标志分子及c-Met下游ERK1/2 MAPK通路相关分子表达，观察DCP可否经c-Met通路调控EMT促肝癌侵袭转移；其次通过体内、外实验观察蟾皮通过DCP-c-Met通路调控EMT抑制肝癌侵袭转移的分子机制。通过本课题的实施，将明确蟾皮抑制肝癌侵袭转移的作用机制，对推动中药抗肝癌侵袭转移药物的研究具有重要意义。

目前原发性肝癌（HCC）的主要治疗方法包括手术切除、肝动脉栓塞、化疗、 肝移植等，但由于其具有转移能力强、易复发且对化疗不敏感等特点，导致预后极差。靶向治疗逐渐成为治疗肝癌转移、复发的热点，因此，深入探寻肝癌治疗的靶点及相关信号通路具有重要意义。上皮细胞-间充质转化（EMT）是肿瘤侵袭转移的关键途径，c-Met是抗肿瘤侵袭转移的重要靶点，前期发现脱-γ-羧基凝血酶原（DCP）是激活c-Met信号通路的关键因素，中药蟾皮能抑制DCP诱导的肝癌细胞增殖及侵袭转移，然分子机制尚不明确。本项目首先通过沉默c-Met基因的表达验证了该基因是肝癌侵袭转移过程中的重要靶点，干扰其表达对抑制肝癌发生发展具有重要意义；然后，利用体内外实验探讨了蟾皮在抑制肝癌细胞侵袭转移方面的分子机制，发现蟾皮可通过干预c-Met-MEK-ERK信号通路调控肝癌细胞EMT进而抑制其侵袭转移。总之，本研究提示蟾皮可能作为 c-Met 抑制剂发挥抗肝癌作用，对推动中药抗肝癌侵袭转移机制的研究及新型c-Met抑制剂的研发具有重要意义。