花青素协同奥沙利铂对人结肠癌HCT116细胞增殖及凋亡的影响

目的探究花青素协同奥沙利铂对人结肠癌HCT116细胞增殖及凋亡的机制。方法体外培养人结肠癌HCT116细胞,并分为:对照组、花青素组、奥沙利铂组、联合组;采用MTT法检测花青素、奥沙利铂单药及联合使用对人结肠癌HCT116细胞的增殖情况;采用克隆形成实验检测细胞生长情况;采用流式细胞仪检测各组细胞凋亡情况;采用Western blot检测TGF-βsmad信号通路相关蛋白及增殖凋亡蛋白表达情况。结果由MTT实验发现,花青素对人结肠癌HCT116细胞48 h的半数致死浓度(IC50)为100 g/L,奥沙利铂人结肠癌HCT116细胞48 h的IC50为0.01 g/L,两者联合使用其半数致死浓度显著下降为60 g/L和0.6 g/L。相比对照组,花青素组和奥沙利铂组克隆形成率、Ki67蛋白相对表达、Bcl-2蛋白相对表达水平显著降低(P<0.01),人结肠癌HCT116细胞凋亡率、Bax蛋白相对表达、Smad2和p-Smad2蛋白相对表达、Smad3和p-Smad3蛋白相对表达、TGF蛋白相对表达均显著升高(P<0.01);相比花青素组和奥沙利铂组,联合组克隆形成率、Ki67蛋白相对表达、Bcl-2蛋白相对表达水平显著降低(P<0.01),人结肠癌HCT116细胞凋亡率、Bax蛋白相对表达、Smad2和p-Smad2蛋白相对表达、Smad3和p-Smad3蛋白相对表达、TGF蛋白相对表达均显著升高(P<0.01)。结论花青素协同奥沙利铂可以促进人结肠癌HCT116细胞的凋亡并抑制其增殖,且作用效果显著优于使用单一药物,其作用机制可能是通过调节TGF-β/Smad信号通路实现。

Effects of anthocyanin combined with oxaliplatin on proliferation and apoptosis of human colon cancer HCT116 cells

Objective To explore effect mechanism of anthocyanin combined with oxaliplatin(L-OHP)on proliferation and apoptosis of human colon cancer HCT116 cells.Methods Human colon cancer HCT116 cells were cultured in vitro,and they were divided into control group,anthocyanin group,L-OHP group and combination group.MTT was applied to detect effects of anthocyanin,L-OHP and their combination on proliferation of human colon cancer HCT116 cells.Colony formation assay was applied to detect cell growth.Flow cytometry was applied to detect cell apoptosis in each group.Western blot was applied to detect expression of TGF-βsmad signal pathway related proteins and proliferation-apoptosis proteins.Results MTT found that 48 h 50%inhibitory concentrations(IC50)of anthocyanin and L-OHP for human colon cancer HCT116 cells were 100 g/L and 0.01 g/L,respectively.IC50 of the two combination was significantly reduced to 60 g/L and 0.6 g/L,respectively.Compared with control group,colony formation rate,relative expression levels of Ki67 and Bcl-2 proteins were significantly decreased in anthocyanin group and L-OHP group(P<0.01),while apoptosis rate of human colon cancer HCT116 cells,relative expression of Bax,Smad2,p-Smad2,Smad3,p-Smad3 and TGF proteins were significantly increased(P<0.01).Compared with anthocyanin group and L-OHP group,colony formation rate,relative expression levels of Ki67 and Bcl-2 proteins were significantly decreased in combination group(P<0.01),while apoptosis rate of human colon cancer HCT116 cells,relative expression of Bax,Smad2,p-Smad2,Smad3,p-Smad3 and TGF proteins were significantly increased(P<0.01).Conclusion Anthocyanin combined with L-OHP can promote apoptosis and inhibit proliferation of human colon cancer HCT116 cells,whose effects are significantly better than those of single drug.The action mechanism may be achieved by regulating TGF-β/Smad signaling pathway.