虎杖苷通过蛋白激酶B信号通路影响胃癌细胞增殖凋亡

Polydatin affects the proliferation and apoptosis of gastric cancer cells through Akt signaling pathway

目的研究虎杖苷对胃癌细胞SGC-7901增殖和凋亡的影响和机制。方法虎杖苷处理胃癌细胞SGC-7901,MTT法检测增殖,平板克隆实验检测克隆形成能力,碘化丙啶(PI)单染法检测细胞周期分布,膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/PI)双染法检测细胞凋亡,蛋白质印迹法(Western blotting)检测细胞周期蛋白D1(cyclin D1)、周期蛋白依赖性激酶4(CDK4)、Bcl-2相关X(Bax)、剪切型胱天蛋白酶-3(Cleaved-caspase-3)、磷酸化-蛋白激酶B(p-Akt)蛋白表达。用蛋白激酶B(Akt)信号激活剂和虎杖苷联合处理胃癌细胞SGC-7901,检测细胞增殖、克隆、周期、凋亡变化。结果虎杖苷处理以后的胃癌细胞SGC-7901增殖能力下降[(0.58±0.06)比(0.20±0.01)],细胞克隆形成数目减少[(118.54±10.65)个比(69.52±7.91)个],细胞凋亡增多[(2.97±0.32)%比(17.45±1.69)%],细胞G0/G1比例升高[(50.47±3.25)%比(67.13±3.84)%],cyclin D1、CDK4蛋白表达减少,Bax、Cleaved-caspase-3蛋白表达水平升高,p-Akt蛋白水平降低[(0.51±0.05)比(0.24±0.03)]。Akt信号激活剂处理可以逆转虎杖苷对胃癌细胞SGC-7901增殖、克隆抑制和周期阻滞、凋亡促进作用。结论虎杖苷通过抑制Akt信号通路阻碍胃癌细胞增殖并诱导细胞凋亡。

Objective To study the effect and mechanism of polydatin on proliferation and apoptosis of gastric cancer cell line SGC-7901.Methods Polydatin was used to treat gastric cancer cell SGC-7901,MTT was used to detect proliferation,and plate cloning experiment was used to detect clone formation ability,cell cycle distribution was detected by PI single staining,apoptosis was detected by Annexin V-FITC/PI double staining,and cyclin D1,CDK4,Bax,Cleaved-caspase-3 and p-Akt protein were detected by Western blotting.Gastric cancer cell line SGC-7901 was treated with Akt signaling activator combined with polydatin,and cell proliferation,cloning,cycle and apoptosis were detected.Results After treatment with polydatin,the proliferation of gastric cancer cells SGC-7901 decreased[(0.58±0.06)vs.(0.20±0.01)],the number of cell clones decreased[(118.54±10.65)vs.(69.52±7.91)],the apoptosis increased[(2.97±0.32)%vs.(17.45±1.69)%],the ratio of G0/G1 increased[(50.47±3.25)%vs.(67.13±3.84)%],the expression of cyclin D1 and CDK4 decreased,and the expression of Bax and Cleaved-caspase-3 elevated,p-Akt protein level were reduced[(0.51±0.05)vs.(0.24±0.03)].Akt signal activator treatment could reverse the proliferation,clonal inhibition,cell cycle arrest and apoptosis promotion of polydatin in gastric cancer cell line SGC-7901.Conclusion Polydatin inhibits proliferation of gastric cancer cells and induces apoptosis by inhibiting Akt signaling pathway.