五倍子单宁酸体外诱导人膀胱癌EJ细胞凋亡及机制研究

Mechanism study of tannin acid induces apoptosis of human bladder cancer EJ cells in vitro

目的观察五倍子单宁酸(TA)在体外培养条件下对人膀胱癌EJ细胞增殖、凋亡的影响及其作用机制。方法将EJ细胞分为阴性对照组(TA浓度为0μmol/L)和不同梯度浓度TA实验组,阴性对照组加入不含TA药物的培养基,其他各组加入含有梯度浓度TA的培养基;用台盼蓝染色检测细胞损伤率,CCK-8检测细胞增殖率,吖啶橙/溴化乙锭(AO/EB)双荧光染色以及流式分析检测细胞凋亡率,Western Blot(WB)检测细胞内半胱氨酸天冬氨酸蛋白酶-3(Caspase-3),BCL2-AssociatedX蛋白(Bax),B淋巴细胞瘤-2(Bcl-2)蛋白表达。结果与阴性对照组比较,实验组TA可降低EJ细胞活力,抑制细胞增殖,诱导细胞凋亡,促进细胞内Caspase3表达、降低Bcl-2蛋白表达、增加Bax/Bcl-2比值差异均具有统计学意义(P<0.05)。结论TA可能通过调节细胞内Caspase3、Bax、Bcl-2蛋白表达诱导EJ细胞凋亡,抑制增殖,提示TA可能是一种潜在的抗膀胱癌药物。

Objective To observe the effect of tannic acid(TA)on the proliferation and apoptosis of human bladder cancer EJ cells in vitro and explore its mechanism.Methods EJ cells were divided into negative control group(0μmol/L)and different concentrations TA groups.In addition to the negative control group,the other groups were added content corresponding medicaments intervene.Cell viability was detected by trypan blue staining,CCK-8 assay was used to detect cell proliferation rate acridine orange/ethidium bromide(AO/EB)dual fluorescence staining and flow cytometry were used to detect apoptosis of EJ cells;the expression of Caspase-3,Bax,and Bcl-2 was detected by Western Blot.Results Compared with negative control group,TA in experimental groups could reduce EJ cell viability inhibit cell proliferation,induce apoptosis,promote Caspase-3 in tracellular expression,decrease expression of Bcl-2 protein,increase Bax/Bcl-2 ratio,the differences were statistically significant(P<0.05).Conclusion TA can promote apoptosis,inhibit cell proliferation and decrease cell viability of human bladder cancer EJ cells,which may be related to its regulation of Caspase-3,Bax,Bcl-2 protein expression to promote apoptosis of EJ cells.These results suggest that gallnut TA may be a potential anti bladder cancer drug.