Sorcin表达受抑乳腺癌多药耐药细胞株MCF-7/A02对化疗药物敏感性的观察

摘要

目的 观察可溶性耐药相关钙结合蛋白基因( Sorcin)表达受抑乳腺癌多药耐药细胞株MCF-7/A02对化疗药物敏感性的影响,并探讨其机制.方法 将MCF-7/A02细胞分为两组,观察组转染针对Sorcin基因的siRNA,对照组不转染.MTT法检测细胞对阿霉素(ADR)、依托泊苷(VP-16)、高三尖杉酯碱(HHT)、长春新碱(VCR)的敏感性,用流式细胞仪检测细胞对碱性蕊香红-123(Rho-123)的外排功能及VP16处理后的细胞凋亡率,Western blot 法检测细胞中的Bcl-2、Bax蛋白.结果 观察组ADR、VP-16、HHT、VCR的IC50分别为(19.92+1.29)、(31.47±2.36)、(4.19±0.27)、(2.96±0.23) μg/ml,对照组分别为(111.57±5.60)、(144.77±20.43)、(24.30±0.43)、(5.89±0.28)μg/ml,两组比较,P均＜0.01.观察组细胞凋亡率为9.15%±0.25%,对照组为4.10%±0.12%,两组比较,P＜0.01.观察组细胞内Rho-123的荧光强度为6.34±0.86,对照组为5.42±0.73,两组比较,P＞0.05.观察组细胞中Bcl-2、Bax蛋白表达量为0.59±0.032、1.80±0.03,对照组分别为0.69±0.025、1.94±0.04,两组比较,P均＜0.05.结论 抑制Sorcin表达可显著增强MCF-7/A02细胞对化疗药物的敏感性；这种作用可能与其调节Bcl-2/Bax通路促进MCF-7/A02细胞凋亡有关.%Objective To study the effect of soluble resistance-related calcium-binding protein gene ( Sorcin) on mul-tidrug resistance (MDR) in MCF-7/A02 cells, and explore the correlative mechanisms. Methods MCF-7/A02 cells were divided into observe group (transfected with Sorcin gene siRNA) and control group (non-transfection). The expression of Sorcin and proteins of Bcl-2, Bax were measured by western blot after siRNA transfection in MCF-7/A02 cells. The sensitivity of cells to ADR, VP-16, HHT and VCR were measured by MTT assays. The accumulation of cytotoxic agents in cells and the apoptotic rate to VP-16 after siRNA transfection were detect by FACS. Results The ADR, VP-16, HHT, VCR IC50 of the observe group were (19.92 ± 1.29) ,(31.47 ±2. 36) , (4. 19 ±0. 27), (2. 96 ±0. 23) fig/ml, control group were (111.57 ±5.60),(144.77 ±20.43),(24. 30 ±0.43), (5. 89 ±0. 28) μg/ml, all P<0.01. Apoptosis rate of the treatment group was 9. 15% ±0.25% , control group was 4. 10% ±0. 12% , P <0.01. The cells MFI of observe group was 6. 34 ±0. 86, control group was 5. 42 ±0.73, P >0.05. Bcl-2/GAPDH, Bax/GAPDH light density ratio of the treatment group was 0.59 ±0.03 and 1. 80 ± 0.03,control group was 0.69 ± 0.20 and 1. 94 ± 0.04, all P <0. 05. Conclusion The reduction of the expression of Sorcin leads to partly reversion of MDR phenotype in MCF-7/A02 cells, which is associated with the changes of Bcl-2 and Bax expression.