摘要
目的探讨姜黄素和Wortmannin联用对胃癌细胞增殖凋亡的影响及机制。方法培养人胃癌SGC-7901细胞,CCK8实验检测10μmol/L、20μmol/L、40μmol/L、80μmol/L、160μmol/L姜黄素和10 nmol/L、50 nmol/L、100 nmol/L、500 nmol/L、1 000 nmol/L的Wortmannin分别作用于细胞24 h、48 h、72 h的细胞增殖情况,计算半数抑制浓度(IC50);根据IC50选择最佳的姜黄素及Wortmannin作用浓度;将接下来的试验分为对照组、姜黄素组、Wortmannin组、姜黄素+Wortmannin组,CCK8试验检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测Cleaved caspase3、PI3K、p-AKT蛋白表达。结果不同浓度的姜黄素和Wortmannin处理细胞24 h、48 h、72 h后的细胞存活率均显著低于0 h细胞存活率(P<0.05或P<0.01),选择30μmol/L姜黄素和170 nmol/L的Wortmannin做后续研究。姜黄素组、Wortmannin组、姜黄素+Wortmannin组细胞存活率及PI3K、p-AKT蛋白表达显著低于对照组,细胞凋亡率及Cleaved caspase3蛋白表达显著高于对照组(P<0.01);姜黄素+Wortmannin组细胞存活率及PI3K、p-AKT蛋白表达显著低于姜黄素组和Wortmannin组,胞凋亡率及Cleaved caspase3蛋白表达显著高于姜黄素组和Wortmannin组(P<0.01)。结论姜黄素和Wortmannin均能抑制人胃癌SGC-7901细胞增殖,姜黄素和Wortmannin联用对细胞增殖凋亡的影响强于单独用姜黄素和Wortmannin。
Objective To investigate the effect and mechanism of curcumin combined with Wortmannin on the proliferation and apoptosis of gastric cancer cell line. Methods Human gastric cancer SGC- 7901 cells were cultured,cell proliferation after 10 μmol / L,20 μmol / L,40μmol /L,80 μmol /L,160 μmol /L of curcumin and 10 nmol /L,50 nmol /L,100 nmol /L,500 nmol /L,1 000 nmol /L of Wortmannin for 24 h,48 h,72 h were detected by CCK8 assay,and IC50 were calculated; according to the IC50,the best concentration of curcumin and Wortmannin were selected and the objects were divided into control group,curcumin group,Wortmannin group,curcumin ＋ Wortmannin group. Cell proliferation was detected by CCK8 test,apoptosis was detected by flow cytometry,the expression of Cleaved Caspase3,PI3 K,p- AKT protein were detected by Western blot. Results The cell survival rate after different concentrations of curcumin and Wortmannin treated cells for 24 h,48 h,72 h was significantly lower than the cells survival rate on 0 h（ P〈0. 05 or P〈0. 01）,30 μmol / L curcuminand and 170 nmol / L Wortmannin were selected for subsequent research. Cell survival rate and PI3 K,p- AKT protein expression in Curcumin group,Wortmannin group,curcumin ＋Wortmannin group were significantly lower than that of the control group,the apoptosis rate and the expression of Cleaved Caspase3 protein was significantly higher than that of the control group（ P〈0. 01）; cell survival rate and PI3 K,p- AKT protein expression in curcumin ＋ Wortmannin group were significantly lower than those of curcumin group and Wortmannin group,cell apoptosis rate and expression of Cleaved Caspase3 protein were significantly higher than those of curcumin group and Wortmannin group（ P〈0. 01）. Conclusion Both curcumin and Wortmannin can inhibit the proliferation of human gastric cancer SGC- 7901 cells,and the effect of curcumin and Wortmannin on the proliferation and apoptosis of human gastric cancer cells are stronger than those of curcumin and Wortmannin.
出处
《临床和实验医学杂志》
2017年第4期335-339,共5页
Journal of Clinical and Experimental Medicine
