南蛇藤提取物靶向mTOR抑制人肝癌HepG2细胞侵袭与转移能力的研究

目的探索南蛇藤提取物(COE)靶向哺乳动物雷帕霉素靶蛋白(m TOR)抑制人肝癌Hep G2细胞增殖、侵袭与转移的分子机制。方法用si RNA技术,构建敲除m TOR基因的Hep G2细胞模型,MTT法检测COE对Hep G2/m TOR-细胞增殖能力的影响。划痕实验及Transwell实验检测药物对细胞侵袭转移能力的影响。Western blotting法分析用药后基质金属蛋白酶-2(MMP-2)、MMP-9蛋白的表达水平变化。结果成功构建敲除m TOR表达的Hep G2/m TOR-细胞。COE明显抑制Hep G2/m TOR-细胞的增殖(P0.05),并呈浓度依赖性。划痕实验结果显示COE降低了细胞迁移能力。Transwell实验结果表明,COE(80 mg/L)明显减少了穿膜细胞数目(P0.05)。Western blotting结果显示,COE(80 mg/L)明显降低MMP-2、MMP-9蛋白的表达水平(P0.05)。结论 COE明显抑制Hep G2/m TOR-细胞的增殖与侵袭转移,m TOR通路可能是其抑制肝癌的潜在作用靶点之一。

Celastrus orbiculatus extracts inhibit invasion and metastasis in human hepatocellular carcinoma HepG2 cells by targeting mTOR

Objective To investigate the molecular mechanisms of Celastrus orbiculatus extracts (COE) of the invasion and metastasis inhibition in human hepatocellular carcinoma HepG2 cells by targeting mTOR. Methods The HepG2/mTOR^- cells with mTOR knockout expression were constructed by using siRNA technology. The effect of COE on the proliferation of the HepG2/mTOR^-cells was also studied. The HepG2/mTOR^- cells were treated with COE in different concentrations (20, 40, 80, 160, and 320 mg/L) for 24 h. The cell reproductive capability of HepG2/mTOR^- cells was detected by MTT. The effect of COE on the metastatic ability of HepG2/mTOR^- cells in vitro was investigated by scratch assay and Transwell migration assay. The expression levels of molecular mechanisms related proteins MMP-2 and MMP-9 were assessed by Western blotting. Results The HepG2/mTOR^- cells with mTOR knockout expression were successfully constructed. COE significantly inhibited the proliferation of HepG2/mTOR^- cells in a concentration-dependent manner (P < 0.05). COE decreased the invasion and migration of HepG2/mTOR^- cells. The results of Transwell experiment indicated that COE (80 mg/L) significantly reduced the number of transmembrane cells (P < 0.05). And the expression levels of MMP2 and MMP9 protein were significantly reduced in the HepG2/mTOR^- cells after the treatment of COE. Conclusion COE can significantly inhibit the proliferation, invasion, and migration in the HepG2/mTOR^- cells. Our data reveal that COE is a potential chemotherapeutic drug in human hepatocellular carcinoma treatments via targeting mTOR signal pathway.