半夏泻心汤通过调控PI3K/AKT/mTOR信号通路对结肠癌细胞增殖和转移的影响

Banxia Xiexin Decoction Inhibit Proliferation and Metastasis of Colon Cancer Cell by Regulating PI3K/AKT/mTOR Signaling Pathway

目的:观察半夏泻心汤调控磷脂酰肌醇-3激酶(phosphoinositide 3-kinases,PI-3K)/蛋白激酶B(protein kinase B,AKT)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路对人结肠癌HT-29细胞增殖和转移的作用机制。方法:将40只大鼠按随机数字表法分为4组,每组10只,分别是空白对照组、半夏泻心汤+5-氟尿嘧啶(5-florouracil,5-FU)组、5-FU组和半夏泻心汤组。采用MTT法检测各组大鼠用药后对人结肠癌HT-29细胞增殖的影响;采用划痕实验检测各组大鼠用药后对HT-29细胞转移能力的影响;采用Western-blotting法检测各组用药对大鼠mTOR、PTEN蛋白表达水平的影响。结果:半夏泻心汤+5-FU组能够抑制HT-29细胞增殖,且呈剂量-时间依赖性,与其他3组比较,差异均有统计学意义(P<0.05);半夏泻心汤+5-FU组能够抑制HT-29细胞转移,且呈剂量依赖性,与其他3组比较,差异均有统计学意义(P<0.05);半夏泻心汤+5-FU组上调抑癌基因PTEN蛋白表达量,能够下调mTOR蛋白表达量,与其他3组比较,差异均有统计学意义(P<0.05)。结论:半夏泻心汤具有抑制人结肠癌HT-29细胞增殖和转移的作用,其机制可能与调控PI3K/AKT/mTOR信号通路有关。

Objective：To investigate the action mechanism of Banxia Xiexin Decoction in regulating of phosphoinositide 3-kinases （ PI-3 K）/protein kinase B （AKT）/mammalian rapamycin target protein （mammalian） by Target of rapamycin, roTOR） of mTOR signaling pathway on proliferation and metastasis of human colon cancer HT-29 cells. Methods ：40 rats were divided into 4 groups randomly, with 10 rats in each group. They were blank control group, Banxia Xiexin Decoction plus 5-fluorouracil group,5-fluorou- racil group and Banxia Xiexin Decoction group. The effect of Banxia Xiexin Decoction on the proliferation of human colon cancer HT-29 cells was detected by MTI＇ assay. The effect of Banxia Xiexin Decoction on the metastatic ability of HT-29 cells was detec- ted by scratch test. The levels of mTOR of PTEN were detected by using Western-blotting method. Results：Banxia Xiexin Decoc-tion plus 5-fluorouracil could inhibit the proliferation of HT-29 cells in a dose-time-dependent manner, and the difference was sta- tistically significant （ P 〈 0.05 ）. Banxia Xiexin Decoction plus 5-fluorouracil could inhibit the metastasis of HT-29 ceils in a dose- time-dependent manner, and the difference was statistically significant （P 〈 0.05 ）. Banxia Xiexin Decoction could down-regulate the expression of mTOR protein and up-regulate the expression of tumor suppressor gene PTEN protein, and the difference was sta- tistically significant （P 〈 0.05 ）. Conclusion ： Banxia Xiexin Decoction can inhibit the proliferation and metastasis of human colon cancer HT-29 cells, and its mechanism may be related to the regulation of PI3K/AKT/mTOR signaling pathway.