信号转导与转录激活子3低表达影响宫颈癌HeLa细胞的超微结构及化疗药物敏感性的初步研究

Knock-down of signal transducers and activators of transcription 3 in cervical cancer cells mediated changes in ultrastructure and the sensitivity to chemotherapy

目的敲低宫颈癌HeLa细胞中信号转导与转录激活子3(signal transducers and activators of tran-scription 3,STAT3)的表达,观察肿瘤细胞超微结构变化,并检测化疗药物诱导肿瘤细胞凋亡的情况。方法利用在线设计软件小分子干扰核糖核酸(small interfering RNA,siRNA)Target Finder设计一对STAT3短发夹RNA(short hairpin RNA,shRNA)的上下游引物,在宫颈癌HeLa细胞中特异性敲低STAT3的表达,通过透射电子显微镜观察对照组及STAT3敲低组HeLa细胞内染色体分布情况,并利用流式细胞术检测化疗药物多柔比星诱导对照组及STAT3敲低组细胞凋亡情况的差异。结果免疫印记表明,STAT3特异性shRNA可在HeLa细胞中显著敲低STAT3的表达;电镜观察发现,在HeLa细胞中敲低STAT3表达可显著诱导染色体边集现象;流式细胞术检测表明,敲低STAT3表达可显著增强多柔比星诱导的细胞凋亡。结论敲低STAT3表达可诱导宫颈癌细胞发生超微结构的改变,并增强宫颈癌细胞对化疗药物的敏感性。

ObjectiveTo evaluate the role of signal transducers and activators of transcription 3 （STAT3） in the regulation of ultrastructure and sensitivity to chemotherapy of cervical cancer cells, STAT3 was knocked down in cervical cancer cells. MethodsTo knockdown STAT3 expre ssion in cervical cancer cells, the online software, small interfering RNA （siRNA） Target Finder, was employed to design STAT3 specific short hairpin RNA （shRNA）. The distribution of chromatin in the HeLa cells transfected by control shRNA or STAT3 specific siRNA was evaluated by transmi ssion electron microscope. To test the role of STAT3 in the sensitivity to Doxorubicin of cervical cancer cells, the apoptosis of cervical cancer cells expressing in control siRNA and STAT3 siRNA were tested by flow cytometry.ResultsEvident down regulation of STAT3 expression in the HeLa cells expression STAT3 shRNA compared to the cells expressing control shRNA was found through Western blotting assay. Chromatin margination was found in most HeLa cells expressing STAT3 shRNA,and knock down of STAT3 in HeLa cells dramatically enhanced the sensitivity to Doxorubicin treatment. ConclusionSilencing STAT3 expression in cervical cancer cells induced changes in ultramicrostructure and enhanced the sensitivity to chemotherapy, implying that STAT3 may be a potential target for the therapy of cervical cancer.