siRNA沉默膀胱癌肝素酶基因对T24细胞增殖及MMP-2表达的影响  

Effects of the siRNA silencing heparanase gene on T24 cells′ proliferation and MMP-2 expression in human bladder cancer

作　　者：张开舒[1] 徐忠华[1] 姜虹[2] 阎磊[1] 周尊林[1] 王荣[2] 刘海南[1] 范医东[1] 刘照旭[1] 郑宝钟[1] 亓天伟[1] 

机构地区：[1]山东大学齐鲁医院泌尿外科,济南250012 [2]教育部和卫生部心血管重构与功能研究重点实验室,济南250012

出　　处：《山东大学学报（医学版）》2008年第6期561-565,570,共6页Journal of Shandong University:Health Sciences

基　　金：山东省科技发展计划资助项目(2005GG4202006)

摘　　要：目的研究siRNA干扰沉默膀胱癌肝素酶基因对T24细胞增殖及MMP-2表达的影响方法体外化学合成肝素酶特异性小干扰RNA(siRNA),用脂质体2000转染至T24膀胱移行细胞癌细胞内,常规分为正常对照组、空白对照组、阴性对照组和转染实验组进行实验,用噻唑蓝(MTT)法检测细胞增殖情况,反转录聚合酶链法(RT-PCR)检测肝素酶基因的表达,免疫细胞化学方法检测MMP-2的表达。结果RNA干扰沉默T24膀胱移行细胞癌细胞肝素酶基因后,T24细胞增殖活力下降,三个不同干扰组细胞活力分别下降了(28.55±3.66)%(、27.20±3.25)%和(37.29±4.82)%。肝素酶mRNA表达下降,膀胱移行细胞癌细胞中MMP-2(明胶酶A)的表达亦明显下降(P均<0.05)。结论RNA干扰肝素酶基因后,细胞活力及肝素酶mRNA表达水平下降并且明显抑制膀胱癌细胞中MMP-2的表达。Objective To explore the changes of proliferation and MMP-2 expression in T24 cells after siRNA down-regulating heparanase （Hpa） gene on bladder cancer cell line-T24. Methods Cells were divided into six groups： the normal control group, the blank control group, the negative control group, the Hpa-siRNA-Ⅰ group, the Hpa-siRNA- Ⅱ group, and the HpasiRNA-Ⅲ group. Chemically synthesized small interfering RNA （siRNA） silencing Hpa gene was transfected into the human bladder cancer cell line-T24. Then proliferation of T24 cells was determined by MTT assay, expression of Hpa mRNA was determined by reverse transcription polymerase chain reaction （RT-PCR）, and expression of MMP-2 was determined by immunocytochemistry. Results MTF showed that the growth of T24 cells was decreased by （28.55 ±3.66） %, （27.20 ± 3.25） % and （37.29±4.82）% in the three experimental groups. RT-PCR analyses also showed that the Hpa mRNA level was significantly decreased in some experimental groups compared with the control groups. Also immunocytochemical staining showed that the MMP-2 （Gelatinase A） expressions were much lower in the experimental group than those in the control groups（ P 〈0.05）. Conclusions RNA interference （RNAi） could significantly decrease T24 cell proliferation and the Hpa gene mRNA level, and inhibits the MMP-2 expression in the bladder cancer cell line-T24.

关 键 词：RNA干扰 肝素裂合酶 明胶酶A 膀胱肿瘤 T24细胞 

分 类 号：R737.14[医药卫生—肿瘤]