siRNA干扰MT1H基因对A549/DDP细胞耐药性的逆转  

Reversal of drug resistance in A549/DDP cells by silencing MT1H gene with MT1H siRNA

作　　者：侯新芳[1] 樊青霞[1] 王留兴[1] 王瑞林[1] 陆士新[2] 赵培荣[1] 

机构地区：[1]郑州大学肿瘤中心郑州大学第一附属医院肿瘤科,郑州450052 [2]中国医学科学院肿瘤研究所病因室,北京100021

出　　处：《肿瘤》2007年第4期277-280,共4页Tumor

摘　　要：目的:探讨siRNA干扰金属硫蛋白1H(metallothionein 1H,MT1H)基因逆转A549/DDP细胞耐药的可行性。方法:采用RT-PCR方法检测MT1H基因在A549和其顺铂耐药株A549/DDP细胞中的表达;将针对MT1H的siRNA导入A549/DDP细胞;用RT-PCR和斑点印迹方法分析MT1H基因表达情况;MTT法观察细胞顺铂耐药性;TUNEL、流式细胞术检测顺铂诱导细胞凋亡率;免疫细胞化学分析凋亡相关蛋白Bcl-2、Bax的表达。结果:MT1H在A549/DDP细胞中高表达但不在A549细胞中表达;A549/DDP细胞转染48 h后,与对照组比较,MT1HsiRNA转染组MT1H mRNA和蛋白表达均明显下调,细胞对DDP的药物敏感性明显提高,DDP诱导凋亡率明显增加,Bcl-2表达明显下降,Bax表达无变化。结论:MT1H基因沉默能降低Bcl-2表达,增强顺铂对A549/DDP细胞凋亡诱导作用,有效逆转A549/DDP细胞耐药。Objective：To explore the feasibility of using siRNA targeting metallothionein 1H （MT1H） gene to reverse the drug resistanee of A549/DDP cells to eisplatin （DDP）. Methods： The expressions of MT1H mRNA in A549 and A549/DDP cells were detected by reverse transeriptase polymerase chain reaction （RT-PCR）. MT1H siRNA was transfeeted into A549/DDP cells with high expression of MT1H. The mRNA and protein levels of MT1H were deteeted by RT-PCR and Dot blotting, respectively. The ehemosensitivity of A549/ DDP eeUs to eisplatin was assessed by MTT assay. The apoptotie ratio induced by DDP was determined by TUNEL and flow eytometry analysis （FCM）. The apoptosis-related protein levels of Bel-2 and Bax were determined by irnmunohistoehemistry. Results： MT1H mRNA was highly expressed in A549/DDP cells but not in A549 cells. The mRNA and protein levels of MT1H were significantly down-regulated at 48 h after transfeetion in MT1H siRNA group compared with the control group. The ehemosensitivity of A549/DDP cells to DDP was significantly elevated. The apoptotie ratio induced by DDP was significantly increased. Expression of Bel-2 was greatly down-regulated but the expression of Bax had little change. Conclusion： Sileneing biT/H gene with MT1H siRNA reduces the expression of Bel-2 , enhanced the apoptosis-indueing effect of DDP on A549/DDP cells, and effectively reverses the drug resistance of A549/DDP cells.

关 键 词：癌 非小细胞肺 RNA 小分子干扰 MTIH基因 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]