摘要
目的探讨抑制BCAR1基因对乳腺癌细胞抗雌激素药物三苯氧胺(tamoxifen,TAM)耐药的影响及作用机制。方法将人类雌激素依赖型乳腺癌细胞株ZR-75-1分为雌二醇(E2)+TAM+空白载体组、E2+TAM+shRNA-β-actin组和E2+TAM+shRNA-BCAR1组。采用RT-PCR和W estern b lot检测各组的抑制效果;4-甲基偶氮四唑蓝(MTT)细胞增殖试验观察TAM对各组乳腺癌细胞生长的影响;流式细胞仪DNA定量法观察TAM作用后各组乳腺癌细胞周期的变化。结果shRNA-BCAR1在mRNA水平和蛋白水平均明显抑制BCAR1基因的表达。BCAR1表达被抑制后,癌细胞对抗雌激素药物TAM的耐药性明显降低,与对照组相比,差异有统计学意义(P<0.05)。结论BCAR1基因高表达与乳腺癌细胞对抗雌激素药物TAM耐药密切相关。
Objective To investigate the effects of BCAR1 inhibited by RNA interference on resistance to antiestrogen drugs tamoxifene in human breast cancer cell in vitro and the mechanisms involved.Methods In vitro the non-carrier、shRNA-BCAR1 and shRNA-β-actin were transfected into ZR-75-1 cells using the agent Lipofectamine.The inhibitory effects of siRNAs was detected by RT-PCR and Western blot;Using the dimethylthiazol-diphehetrazolium bromide（MTT） method to observe the cell proliferation;Cell cycle were analyzed with flow cytometric DNA studies.Results The BCAR1 gene expression could be specifically knocked down by the shRNA-BCAR1 detected with both RT-PCR and Western blot.After the inducible knockdown,the resistance to antiestrogen drugs of the breast cancer cell in BCAR1 group was significantly inhibited compared with that of controls（P〈0.05）.Conclusion Overexpression of BCAR1 is raleted with the drug resistance of breast cancer cells resistance to antiestrogen drugs tamoxifene.
出处
《宁夏医学杂志》
CAS
2008年第10期865-867,共3页
Ningxia Medical Journal
