长春新碱调控PI3K/Akt信号通路抑制肝癌细胞Hep3B增殖迁移侵袭及促进细胞凋亡的机制研究

Vincristine regulates PI3K/Akt signaling pathway to inhibit the proliferation,migration and invasion of hepatocellular carcinoma Hep3B and promote apoptosis

目的:探讨长春新碱对肝癌细胞Hep3B增殖、凋亡、迁移和侵袭的影响及机制。方法:体外培养Hep3B细胞,分为对照组(NC组)、不同浓度(0.02、0.04、0.08μmol/L)长春新碱组、长春新碱+胰岛素样生长因子-1(IGF-1)组,用细胞计数试剂盒-8(CCK-8)法检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell检测细胞迁移和侵袭,蛋白印迹(Western Blot)法检测细胞中Ki-67、活化的半胱天冬酶-3(Cleaved-caspase-3)、基质金属蛋白酶(MMP)2、MMP9、磷酸化磷脂酞肌醇3-激酶(p-PI3K)和磷酸化的蛋白激酶B(p-Akt)的蛋白表达水平。结果:与NC组比较,长春新碱组Hep3B细胞OD450 nm值、迁移和侵袭数及Ki-67、MMP2、MMP9、p-PI3K和p-Akt蛋白表达降低(P<0.05),细胞凋亡率及Cleaved-caspase-3蛋白表达升高(P<0.05)。与长春新碱组比较,长春新碱+IGF-1组Hep3B细胞OD450 nm值、迁移和侵袭数及Ki-67、MMP2、MMP9、p-PI3K和p-Akt蛋白表达升高(P<0.05),细胞凋亡率和Cleaved-caspase-3蛋白表达降低(P<0.05)。结论:长春新碱可能通过抑制PI3K/Akt信号通路降低肝癌细胞Hep3B的增殖、迁移和侵袭能力,并诱导细胞凋亡。

Objective:To investigate the effect mechanism of vincristine on the proliferation,apoptosis,migration and invasion of liver cancer cell Hep3B.Methods:Hep3B cells were cultured in vitro and divided into control group(NC group),vincristine group with different concentrations(0.02,0.04,0.08μmol/L),vincristine+insulin-like growth factoR-1(IGF-1)group,then cell proliferation was detected by CCK-8 method,cell apoptosis was detected by flow cytometry,cell migration and invasion was detected by Transwell,and the protein expression levels of Ki-67,Cleaved-caspase-3,MMP2,MMP9,p-PI3K and p-Akt were detected by Western Blot method.Results:Compared with the NC group,the OD450 nm value,migration and invasion number of Hep3B cells and the protein expression of Ki-67,MMP2,MMP9,p-PI3K and p-Akt in the vincristine group were decreased(P<0.05),while apoptosis rate and the protein expression of Cleaved-caspase-3 were increased(P<0.05).Compared with the vincristine group,the OD450 nm value,migration and invasion number of Hep3B cells and the protein expression of Ki-67,MMP2,MMP9,p-PI3K and p-Akt in the vincristine+IGF-1 group were increased(P<0.05),while apoptosis rate and the protein expression of Cleaved-caspase-3 were decreased(P<0.05).Conclusion:Vincristine may reduce the proliferation,migration and invasion ability of liver cancer cell Hep3B and induce its apoptosis by inhibiting PI3K/Akt signaling pathway.