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

山东大学学报(医学版)

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

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

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

收稿日期:2007-12-22 修回日期:1900-01-01 出版日期:2008-06-16 发布日期:2008-06-16
通讯作者: 徐忠华

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

ZHANG Kai-shu¹, XU Zhong-hua¹, JIANG Hong², YAN Lei¹, ZHOU Zun-lin1,WANG Rong², LIU Hai-nan¹, FAN Yi-dong¹, LIU Zhao-xu¹, ZHENG Bao-zhong¹, QI Tian-wei¹

1. Department of Urology, Qilu Hospital of Shandong University, Jinan 250012, China;2. Key Laboratory of Cardiovascular Remodeling and Function Research

Received:2007-12-22 Revised:1900-01-01 Online:2008-06-16 Published:2008-06-16

摘要/Abstract

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。

关键词: T24细胞, 明胶酶A, 膀胱肿瘤, 肝素裂合酶, RNA干扰

Abstract: 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. MethodsCells 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 Hpa-siRNA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. ResultsMTT 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.

Key words: RNA interference, Heparinase, Gelatinase A, Bladder neoplasms, T24 cells

中图分类号:

R737.14

张开舒,徐忠华,姜虹,阎磊,周尊林,王荣,刘海南,范医东,刘照旭,郑宝钟,亓天伟 . 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[J]. 山东大学学报(医学版), 2008, 46(6): 561-565.

ZHANG Kai-shu,XU Zhong-hua,JIANG Hong,YAN Lei,ZHOU Zun-lin,WANG Rong,LIU Hai-nan,FAN Yi-dong,LIU Zhao-xu,ZHENG Bao-zhong,QI Tian-wei. Effects of the siRNA silencing heparanase gene on T24 cells′proliferation and MMP-2 expression in human bladder cancer[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2008, 46(6): 561-565.

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