四溴苯三唑对宫颈癌HeLa细胞凋亡的诱导作用及其机制

目的：探讨四溴苯三唑（TBB）对人宫颈癌HeLa细胞的促凋亡作用，阐明其对相关信号转导通路的作用机制。方法：选取处于对数生长期的人宫颈癌HeLa细胞，将其分为对照组（未给予TBB）和实验组（给予TBB）。采用MTT比色法检测HeLa细胞存活率，采用倒置显微镜观察人宫颈癌HeLa细胞形态表现，采用Annexin Ⅴ-FITC/PI双染法及流式细胞术检测HeLa细胞凋亡率，采用Western blotting法检测细胞中凋亡相关蛋白p-Akt、Akt、Bcl-2、Bax、cleaved-caspase-3和Pro-caspase-3表达水平。结果：MTT法检测，与对照组比较，实验组HeLa细胞存活率明显降低（P<0.01），且呈浓度依赖性；倒置显微镜下观察，实验组HeLa细胞出现体积缩小、细胞核皱缩等细胞凋亡的形态变化；Annexin Ⅴ-FITC/PI双染法和流式细胞术检测，与对照组比较，实验组各时间段（3、6、12和24 h）细胞凋亡率明显升高（P<0.05或P<0.01）；Western blotting检测，与对照组比较，实验组HeLa细胞中抗凋亡蛋白p-Akt和Bcl-2表达水平逐渐降低，促凋亡蛋白Bax和cleaved-caspase-3表达水平逐渐升高，Pro-caspase-3表达水平逐渐降低（P<0.05或P<0.01）。结论：TBB可能通过抑制Akt信号通路的活性进而有效促进HeLa细胞凋亡。

Apoptotic effect of 4,5,6,7-tetrabromobenzotriazole on human cervical cancer HeLa cells and its mechanism

Objective:To investigate the promotive effect of 4,5,6,7-tetrabromobenzotriazole (TBB) on the apoptosis of the human cervical cancer HeLa cells,and to clarify its effect mechanism in related signaling pathways.Methods:The human cervical cancer HeLa cells at logarithmic growth phase were divided into control group(without TBB) and experiment group(with TBB).MTT assay was used to detect the survival rate of the HeLa cells;the morphology of HeLa cells was observed under inverted microscope;Annexin Ⅴ-FITC/PI double staining and flow cytometry(FCM) were used to determine the apoptotic rates of the HeLa cells;the expression levels of apoptosis-related proteins p-Akt,Akt,Bcl-2,Bax,cleaved-caspase-3,and Pro-caspase-3 were detected by Western blotting method.Results:The MTT results showed that the survival rates of the HeLa cells in experiment group were significantly decreased(P<0.01) in a dose-dependent manner compared with control group.The apoptotic morphology of the HeLa cells in experiment group were found as cell shrinkage and karyopyknosis under inverted microscope.The Annexin Ⅴ-FITC/PI double staining and FCM results showed that the apoptotic rates of the HeLa cells in experiment group (3,6,12 and 24 h) were higher than that in control group (P<0.05 or P<0.01).The Western blotting results showed that compared with control group,the expression levels of the anti-apoptotic proteins p-Akt and Bcl-2 in HeLa cells in experiment group were decreased obviously,whereas the expression levels of the pro-apoptotic proteins Bax and cleaved-caspase-3 were increased and the expression levels of Pro-caspase-3 were decreased.Conclusion:TBB may promote the apoptosis of human cervical cancer HeLa cells by inhibiting the Akt signaling pathway.