干扰HDAC1通过调控STAT3信号通路影响皮肤鳞癌细胞凋亡

Effect of HDAC1 silencing on apoptosis of squamous cell carcinoma of skin via regulating STAT3 signaling pathway

目的:探讨干扰组蛋白去乙酰化酶1(HDAC1)对皮肤鳞癌细胞凋亡的影响。方法:皮肤鳞癌细胞A431分别转染HDAC1小干扰RNA(HDAC1 si RNA)和小干扰RNA阴性对照(si RNA NC),RT-PCR和Western blot检测转染后细胞中HDAC1的表达水平,MTT检测细胞活力,流式细胞术检测细胞凋亡,Western blot检测信号转导及转录激活因子3(STAT3)、磷酸化STAT3(p-STAT3)和cleaved caspase-3的蛋白水平。同时用STAT3信号通路抑制剂作用于转染HDAC1 si RNA的A431细胞,MTT法检测细胞活力,流式细胞术检测细胞凋亡,Western blot检测STAT3、p-STAT3和cleaved caspase-3的蛋白水平。结果:HDAC1 si RNA能够抑制A431细胞中HDAC1的m RNA和蛋白表达。干扰HDAC1表达后细胞活力和细胞中p-STAT3水平下降,而细胞凋亡率和细胞中cleaved caspase-3水平升高。STAT3信号通路抑制剂作用后,转染HDAC1 si RNA的A431细胞活力及p-STAT3水平下降,细胞凋亡率及细胞中cleaved caspase-3水平升高。结论:干扰HDAC1表达可能通过调控STAT3信号通路抑制皮肤鳞癌细胞活力,促进皮肤鳞癌细胞凋亡。

AIM:To investigate the effect of histone deacetylase1(HDAC1)silencing on apoptosis of squamous cell carcinoma of skin.METHODS:Skin squamous cell carcinoma A431cells were transfected with HDAC1small interfering RNA(HDAC1siRNA)or small interfering RNA negative control(siRNA NC).The expression levels of HDAC1in transfected cells were detected by RT-PCR and Western blot.The cell viability was measured by MTT assay,and the apoptosis was analyzed by flow cytometry.The protein levels of STAT3,p-STAT3and cleaved caspase-3were determined by Western blot.The inhibitor of STAT3signaling pathway was used to treat the A431cells transfected with HDAC1siRNA.The cell viability was detected by MTT assay,the apoptosis was analyzed by flow cytometry,and the protein levels of STAT3,p-STAT3and cleaved caspase-3were determined by Western blot.RESULTS:HDAC1siRNA inhibited the expression of HDAC1at mRNA and protein levels in the A431cells.After interfering with the expression of HDAC1,the cell viability and the protein level of p-STAT3in the cells decreased,while the apoptotic rate and the protein level of cleaved caspase-3in the cells were increased.After treatment with the inhibitor of STAT3pathway,the viability of A431cells transfected with siRNA and the protein level of p-STAT3decreased,while the apoptotic rate and the protein le-vel of cleaved caspase-3in the cells were increased.CONCLUSION:Interference with HDAC1expression may regulate the STAT3signaling pathway to inhibit the viability of skin squamous cell carcinoma cells,thus promoting the apoptosis of squamous cell carcinoma of skin.