三羟异黄酮对人乳腺癌耐药细胞mdr-1、HER2/neu表达的影响  

Influence of Genistein on mdr-1 and HER2/neu Gene Expression in MCF-7/ADM Cells

作　　者：王耕[1] 黄韬[1] 薛家鹏[2] 王明华[2] 惠震[2] 

机构地区：[1]华中科技大学同济医学院附属协和医院普外科,武汉430022 [2]湖北医药学院附属太和医院内分泌血管外科,十堰442000

出　　处：《华中科技大学学报（医学版）》2011年第5期589-592,610,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的研究三羟异黄酮(Genistein,GEN)对体外培养人乳腺癌MCF-7/ADM耐药细胞中mdr-1、HER2/neu基因及蛋白表达的影响,探讨其逆转多药耐药机制。方法采用MTT法检测GEN作用MCF-7/ADM细胞后的细胞增殖抑制率(IR)和半数抑制浓度(IC50);RT-PCR法检测MCF-7/ADM细胞经不同浓度阿霉素(ADM)、GEN及GEN联合ADM处理后,其mdr-1、HER2/neu mRNA表达的变化;Western blot检测其对c-erbB2蛋白及P-糖蛋白(P-gp)的影响。结果 GEN对MCF-7/ADM细胞生长抑制作用明显,其抑制效应呈时间-剂量依赖性,特别是GEN浓度增加到60μg/mL时,对细胞抑制作用急剧升高(P<0.01)。MCF-7/ADM细胞中有mdr-1、HER2/neu过量表达,经GEN单独及联合ADM作用后,HER2/neu mRNA表达明显下调,c-erbB2蛋白出现一致性的表达下调,但对mdr-1mRNA及P-gp无影响。结论人乳腺癌MCF-7/ADM细胞耐药性与耐药基因及原癌基因的高表达相关,GEN能下调MCF-7/ADM细胞HER-2/neu基因及蛋白表达,可能是其逆转MCF-7/ADM细胞多药耐药性的机制之一,但对由P-gp介导的多药耐药无效。Objective To study the influence of Genistein（GEN）on multidrug resistance gene-1（mdr-1）and HER2/neu gene and protein expression in MCF-7/ADM cells in vitro,and the reversal mechanism of multidrug resistance. Methods MTT method was employed to detect the inhibition rate（IR）and 50% inhibition concentration（IC50）of GEN on MCF-7/ADM cells.RT-PCR was employed to detect the influence of GEN on the expression of mdr-1 and HER2/neu in MCF-7/ADM cells.By using Western blot,the effect of GEN on c-erbB2 protein and P-gp was examined. Results GEN effectively inhibited the growth of MCF-7/ADM cells in a time-and dose-dependent manner.The inhibitory effect increased dramatically（P0.01）especially when GEN concentration rose to 60 μg/mL.There was an overexpression of mdr-1 and HER2/neu in MCF-7/ADM cells.After treatment with GEN alone or GEN combined with adriamycin,HER2/neu gene expression was significantly decreased,and c-erbB2 protein was consistently down-regulated,but no significant effects on mdr-1 mRNA and P-gp were found. Conclusion The drug resistance of MCF-7/ADM cells is related with the high expression of resistance gene and proto-oncogene.GEN can partly reverse the multidrug resistance of MCF-7/ADM cells probably by down-regulating HER2/neu gene and protein expression in MCF-7/ADM cells,but it is invalid to the P-gp-mediated multidrug resistance.

关 键 词：三羟异黄酮 乳腺癌 MCF-7/ADM 基因 mdr-1 基因 HER2/neu 

分 类 号：R737.9[医药卫生—肿瘤]