敲低B-细胞淋巴瘤/白血病-2原癌基因对子宫颈癌细胞顺铂敏感性的研究

Research on the knockdown of Bcl-2 by siRNA in improving the sensitivity of cervical carcinoma cisplatin sensitivity

目的探讨子宫颈癌C4-1细胞敲低B-细胞淋巴瘤/白血病-2原癌基因(Bcl-2)的表达,提高人子宫颈癌C4-1细胞对顺铂(DDP)化疗敏感性的作用与机制。方法应用敲低Bcl-2慢病毒转染子宫颈癌C4-1细胞,将其分为对照组、空载组及敲低组,上述分组通过DDP处理后分别作为对照+DDP组、空载+DDP组和敲低+DDP组。采用RT-PCR法检测C4-1细胞Bcl-2 mRNA表达水平,采用流式细胞仪检测C4-1细胞凋亡率,四甲基偶氮唑蓝(MTT)比色法检测C4-1细胞对DDP的敏感性。结果敲低组与空载组及正常对照组比较,C4-1细胞的凋亡率明显增加(P<0.05),C4-1细胞Bcl-2 mRNA表达量降低;敲低组+DDP组与空载+DDP组和对照+DDP组比较IC50值降低,上述差异均有统计学意义(P<0.05)。结论慢病毒转染子宫颈癌C4-1细胞敲低Bcl-2表达可增加凋亡率,提高子宫颈癌C4-1细胞对顺铂的敏感性。

Objective To explore the expression of the knockdown of B-cell lymphoma/leukemia-2（Bcl-2） in improving the sensitivity of cervical carcinoma C4-1 cells to cisplatin chemotherapy.Methods Human cervical cancer C4-1 cells were transfected with knockdown of Bcl-2.The cells were divided into the control group,light group and knockdown group,which were treated with DDP,regarded as control ＋ DDP group,light ＋ DDP and knockdown ＋ DDP groups respectively.Real time fluorescent quantitative PCR（q PCR） was used to detect mRNA expression of Bcl-2 gene,and flow cytometry was used to detect cell apoptosis rate,while methyl thiazolyl tetrazolium（MTT） colorimetry was used to detect their sensitivity to DDP.Results Compared with light group and control group,the apoptosis rate of C4-1 cells in knockdown group increased,and the difference of them had statistical significance（P〈0.05）,Bcl-2 mRNA expression of C4-1 cells decreased,and the difference of them had statistical significance（P〈0.05）,and the difference of them had statistical significance（P〈0.05）.Compared with light ＋ DDP group and control ＋ DDP group,IC50 of knockdown ＋ DDP group reduced,and the difference of them had statistical significance（P〈0.05）.Conclusion The slow virus by siRNA build trasfection cervical carcinoma C4-1 cells can knockdown Bcl-2,and increase the apoptosis rate,and improve the sensitivity of cervical cancer C4-1 cells to cisplatin.