人参皂苷Rg3抑制肝癌细胞侵袭迁移的作用靶点研究

Liver cancer as one of common malignancies is characterized by high invasion and migration, which results in high mortality. Our previous studies displayed that the expression of ARHGAP9 was lower in the liver cancer tissue, and the prognosis of low ARHGAP9 expression patients was poor. High expression of ARHGAP9 also inhibited the activation of signalings associated with invasion and migration in liver carcinoma. In vitro experiments showed that low concentration of ginsenoside Rg3 could inhibit the invasion and migration of hepatoma cell but not reduce the cell viability, and promote the expression of ARHGAP9 both in mRNA and protein levels; after introduction of siRNA targeting ARHGAP9, the anti-invasion and migration ability of Rg3 was reduced significantly, indicating that Rg3 may inhibit the invasion and migration of hepatoma cell by bounding to ARHGAP9 protein. In this project, we aim to screen and substantiate definitely the binding target of Rg3 for inhibiting the invasion and migration of hepatoma cell and to investigate the mechanism of inhibiting invasion and migration based on this target by means of cell thermal shift assay, analysis of confocal fluorescence, RNA interference, gene transfection, Q-RT-PCR and Western blot. This research will provide novel molecular target for the treatment of liver cancer, new strategy for traditional Chinese medicine prevention and treatment of liver cancer and new sights for the drug development of liver cancer.

肝癌是最常见的恶性肿瘤之一，高侵袭转移是其重要的生物学特性，是导致病人死亡的重要原因。我们前期研究发现ARHGAP9基因在肝癌组织中低表达，低表达者预后较高表达者差，且高表达会抑制肝癌组织样本中与肝癌转移密切相关的信号通路。体外实验显示低浓度的人参皂苷Rg3能显著抑制肝癌细胞的侵袭迁移但不降低细胞的活力，且能明显促进ARHGAP9基因和蛋白的表达；RNA干扰ARHGAP9后，Rg3抑制肝癌细胞侵袭迁移的能力显著降低，提示Rg3抑制肝癌细胞侵袭迁移的作用靶点可能为ARHGAP9蛋白。本项目拟采用细胞热转移分析技术，结合激光共聚焦荧光分析、RNA干扰、基因转染、Q-RT-PCR和Western blot等方法，确证Rg3抑制肝癌细胞侵袭迁移的作用靶点以及基于该靶点的作用机制，为肝癌治疗提供新的分子靶点，为中药防治肝癌提供新策略，为开发抗肝癌新药提供新思路。

肝癌是最常见的恶性肿瘤之一，高侵袭转移是其重要的生物学特性，是导致病人死亡的重要原因。我们前期研究发现ARHGAP9基因在肝癌组织中低表达，低表达者预后较高表达者差，且高表达会抑制肝癌组织样本中与肝癌转移密切相关的信号通路。体外实验显示低浓度的人参皂苷Rg3能显著抑制肝癌细胞的侵袭迁移，且能明显促进ARHGAP9基因和蛋白的表达。通过细胞热转移分析（CETSA）实验验证ARHGAP9是Rg3的结合靶点；我们研究了ARHGAP9的功能，并发现将ARHGAP9基因干扰后，Rg3未能显著抑制肝癌细胞的侵袭和迁移，表明ARHGAP9是Rg3抑制肝癌细胞侵袭迁移的作用靶点。