STAT3基因表达对宫颈癌HeLa细胞顺铂敏感性的影响

Inhibition of STAT3 enhances the chemosensitivity of human cervical carcinoma cell line HeLa to cisplatin

【目的】探讨信号转导因子与转录激活因子3(signal transduction and activators of transcription 3,STAT3)短发夹RNA(short hairpin RNA,shRNA)真核表达载体对宫颈癌HeLa细胞化疗药物顺铂敏感性的作用。【方法】构建针对STAT3基因的shRNA干扰质粒和阴性对照质粒,同时设空白对照组。采用Lipofectimane 2000转染宫颈癌HeLa细胞,RT-PCR及免疫蛋白印迹法分别检测STAT3基因的mRNA及蛋白表达水平。细胞经不同浓度顺铂(DDP)作用后,流式细胞术(FCS)检测细胞凋亡率,四甲基偶氮唑蓝比色(MTT)法检测细胞生长抑制率并计算DDP的50%抑制浓度(IC50)。【结果】与各对照组细胞相比,转染STAT3基因shRNA真核表达载体的HeLa细胞,STAT3基因在mRNA和蛋白水平表达均下降,凋亡率均明显增加,细胞对DDP的IC50值明显下降,差异均有统计学意义(P<0.05)。【结论】针对STAT3基因的shRNA真核表达载体有效地抑制宫颈癌HeLa细胞STAT3基因表达,增强其对化疗药物DDP的敏感性。

【Objective】To explore the effects of eukaryotic vector expressing short hairpin RNA（shRNA） targeting signal transducers and activators of transcription 3（STAT3） on the chemosensitivity of human cervical carcinoma HeLa cells.【Methods】 Vectors containing shRNA targeting STAT3 or nonspecific gene were constructed,meanwhile an empty control group was designed.The vectors were transfected into HeLa cells with Lipofectimane 200.The mRNA and protein expressions of STAT3 were determined by RT-PCR and Western blotting,respectively.Cells were treated with different concentration of cisplatin（DDP）.The apoptosis rate was measured by flow cytometry（FCS）.The cell viability was detected by methyl thiazolyl tetrazolium（MTT） assay and 50% inhibitive concentration（IC50） of DDP was also examined.【Results】 The STAT3 expression in pSTAT3-siRNA group was significantly inhibited as compared with other groups.At same time,the apoptotic rate was more increased and the IC50 of DDP was more decreased than that of other groups.【Conclusion】 The STAT3 specific shRNA expression vector could effectively suppress the expression level of STAT3 gene in HeLa cells,and enhance their sensitivity to DDP.