紫花牡荆素对宫颈癌HeLa细胞凋亡及DAPK基因甲基化的影响

Effects of casticin on the apoptosis and methylation of DAPK gene in cervical cancer HeLa cells

目的探讨紫花牡荆素(casticin)对人宫颈癌HeLa细胞的凋亡和DAPK基因甲基化以及DAPK蛋白表达的影响。方法用不同浓度的casticin处理体外培养的HeLa细胞;AnnexinV-PI染色流式细胞术定量分析细胞凋亡率;DNA琼脂糖凝胶电泳法观察细胞DNA断裂;基因甲基化特异性PCR检测DAPK基因甲基化状态;Western blotting检测DAPK蛋白表达。结果 casticin可以浓度依赖性的方式有效诱导HeLa细胞凋亡;casticin作用于宫颈癌HeLa细胞48小时后,DAPK基因甲基化程度显著降低;casticin以浓度依赖的方式上调DAPK蛋白表达。结论 casticin可能通过使DAPK基因去甲基化上调DAPK蛋白的表达,进而诱导HeLa细胞凋亡。

Objective To investigate the effects of casticin on the apoptosis,methylation of DAPK gene and expression of DAPK protein in human cervical cancer HeLa cells.Methods Human cervical cancer HeLa cells were treated with different concentrations of casticin,AV-PI staining was conducted to quantitatively assess the rate of apoptosis,DNA agarose gel electrophoresis was used to observe DNA ladder,,methylation-specific PCR was conducted to assess the methylation status of DAPK gene,the expression of DAPK protein was measured by western blot.Results Casticin significantly induced the apoptosis of human cervical cancer HeLa cells in a dose-and time-dependent manner.The apoptosis was observed after treatment with casticin at 8.0μM for 48 h.Demethylation of DAPK gene was occurred by treatment with casticin at the indicated concentrations for 72 hours.The expression of DAPK protein was increased by treatment with casticin,in a concentration dependent manner.Conclusion Casticincan induced apoptosis of human cervical cancer HeLa cells in vitro through demethylation of DAPK gene and upregulation of the expression of DAPK protein.