论文部分内容阅读
目的探讨沉默Ku70基因对耐表阿霉素人乳腺癌细胞MCF-7/ADR耐药逆转作用。方法以脂质体包裹的SiRNA-Ku70转染细胞,RT-PCR验证Ku70沉默效率。CCK8试剂盒测定细胞增殖活性,通过Caspase-3分光光度法、AnnexinⅤ-PI染色检测细胞凋亡。结果 SiRNA-Ku70转染组Ku70-mRNA表达明显下降,细胞对表阿霉素的敏感性明显增加,药物半数致死浓度(IC50)由34.38±6.75μg/ml降低为13.06±2.62μg/ml,耐药指数(RI)由8.26减至3.14。以表阿霉素5μg/ml的培养液培养细胞24h,SiRNA-Ku70转染组(12pmol)与阴性对照组及空白对照组相比,caspase-3活性明显增高;凋亡细胞数明显增多。结论 SiRNA-Ku70通过下调Ku70的表达,逆转耐表阿霉素人乳腺癌细胞MCF-7/ADR对表阿霉素的耐药性,降低凋亡域值,从而增强了人乳腺癌细胞对表阿霉素的敏感性。
Objective To investigate the reversal effect of silencing Ku70 gene on resistance to epirubicin human breast cancer cells MCF-7 / ADR. Methods Liposome-encapsulated SiRNA-Ku70 cells were transfected, and the silencing efficiency of Ku70 was verified by RT-PCR. CCK8 kit assay cell proliferation activity, apoptosis was detected by Caspase-3 spectrophotometry, Annexin Ⅴ-PI staining. Results The expression of Ku70-mRNA in SiRNA-Ku70 transfection group was significantly decreased. The sensitivity of cells to epirubicin was significantly increased. The IC50 decreased from 34.38 ± 6.75μg / ml to 13.06 ± 2.62μg / ml, The drug index (RI) decreased from 8.26 to 3.14. Compared with negative control group and blank control group, the activity of caspase-3 in SiRNA-Ku70 transfection group was significantly increased after culturing cells with epirubicin 5μg / ml for 24h, and the number of apoptotic cells was significantly increased. Conclusion SiRNA-Ku70 can down-regulate the expression of Ku70 and reverse the resistance of epirubicin to epirubicin-resistant human breast cancer cell line MCF-7 / ADR Adriamycin sensitivity.