lncRNA-TTN-AS1竞争性抑制miR-139-5p调控LRH-1表达促肝癌细胞增殖、侵袭及EMT的分子机制及虫草素的干预研究

上皮-间质转化（EMT）在肝癌的侵袭和转移分子机制中起重要作用。肝受体同系物1(LRH-1)可通过增加MMP9表达促肝癌EMT，但调控LRH-1的上游机制不清。前期我们通过高通量检测发现TTN-AS1在肝癌组织中异常高表达，表达量与LRH-1高度相关。预实验中过表达TTN-AS1能促进肝癌细胞增殖、侵袭及EMT，并促LRH-1表达。生物信息预测发现139-5p存在与TTN-AS1及LRH-1的结合位点，且三者在肝癌组织内表达高度相关。结合上一课题发现虫草素能抑制肝癌细胞增殖、侵袭及EMT并下调LRH-1表达，我们推测TTN-AS1通过139-5p调控LRH-1的表达促癌细胞增殖、侵袭及EMT，而虫草素则通过调控LRH-1阻断其促癌作用。本研究拟从分子、细胞和整体水平，在细胞、裸鼠模型内使用细胞挽救实验、荧光免疫实验及免疫共沉淀等方法证实我们的假说，为肝癌侵袭及转移的防治提供新思路及新靶点。

Epithelial-mesenchymal transition (EMT) plays an important role in the molecular mechanism of invasion and metastasis of hepatocellular carcinoma. LRH-1 can increase the expression of MMP9 and promote EMT of hepatoma, but the upstream regulation mechanism of LRH-1 is not clear.In preliminary experiment, we found that the expression of lncRNA-TTN-AS1 was abnormally high in hepatocellular carcinoma, and the expression of LRH-1 was highly correlated with the amount of expression through high-throughput assay. Transfection of the overexpressing TTN-AS1 plasmid could promote the proliferation, invasion, EMT, and LRH-1 has overexpression in hepatocellular carcinoma cells In pre-experiment, suggested that there is a regulatory relationship between them. Through bioinformatics prediction and correlational analysis for hepatocellular carcinoma tissues, miR-139-5p has binding sites with TTN-AS1 and LRH-1. The expression of miR-139-5p has the highest correlation with the expression of TTN-AS1 and LRH-1 in hepatocellular carcinoma. In the previous study, we found that cordycepin could inhibit the proliferation, invasion and could down-regulate expression of LRH-1 in hepatoma cells. We speculate that TTN-AS1 can promote cancer cell proliferation and EMT by regulating the expression of LRH-1 through miR-139-5p, while cordycepin could inhibit tumor promotion accelerated TTN-AS1/miR-139-5p/LRH-1 signal pathway by regulating LRH-1. The purpose of this study is to reveal the carcinogenic effect of TTN-AS1/miR-139-5p/LRH-1 signal pathway and the intervention mechanism of cordycepin by using cell rescue experiment, fluorescence immunoassay and co-immunoprecipitation in molecular, cellular and whole levels, using hepatocellular carcinoma cells and nude mice models, so as to provide new ideas and targets for the prevention of invasion and metastasis of liver cancer.