RNA干扰下调midkine基因表达对人乳腺癌细胞粘附和侵袭力的影响  

Effects of midkine siRNA on adhesion and invasion of human breast cancer cells

作　　者：俞力 范钰[2] 邱志远[2] 周永静[2] 龚丹丹[2] 肖秀娣[2] 武正炎[3] 

机构地区：[1]镇江市分子内分泌实验室,江苏212001 [2]江苏大学附属人民医院肿瘤研究所,212002 [3]江苏省人民医院普外科,210009

出　　处：《中华内分泌外科杂志》2011年第3期148-151,共4页Chinese Journal of Endocrine Surgery

基　　金：镇江市分子内分泌重点实验室项目（SS2009012）：江苏省自然科学基金资助项目（BK2009205）

摘　　要：目的探讨中期因子（midkine，MK）基因siRNA对乳腺癌细胞生物学行为的影响。方法培养人乳腺癌Bcap-37、LCC1、MCF-7、MDA—MB-231、MDA—MB-435、MDA—MB-468及ZR75—1细胞株，以荧光实时定量PCR方法检测MK基因mRNA表达；筛选出MK表达最高者。采用MK siRNA转染乳腺癌细胞株，分别以荧光实时定量RT—PCR和免疫荧光方法观察MK基因mRNA和蛋白水平，然后以四唑蓝（MTT）比色法检测细胞粘附性，以Boyden小室方法检测癌细胞侵袭能力。结果7株乳腺癌细胞中，MK均有不同程度的表达，以MCF-7细胞最高；以MKsiRNA转染乳腺癌MCF-7细胞后，癌细胞MK基因mRNA和蛋白水平明显下降，且呈浓度依赖性；siRNA转染组细胞粘附数量明显下降，细胞侵袭力明显下降，均呈浓度依赖性（P〈0．01；P〈0．01）。结论MK基因在乳腺癌细胞粘附和侵袭中发挥着重要作用；以siRNA转染乳腺癌细胞，可抑制乳腺癌细胞粘附和侵袭能力。Objective To study the effects of midkine （MK） gene small interfering RNA （siRNA） on adhesion and invasion of human breast cancer cells. Methods Real time PCR was used to evaluate MK mRNA expression in 7 human breast cancer cell lines Bcap-37, LCCl, MCF-7, MDA-MB-231, MDA-MB-435, MDAMB-468, and ZR75-1. The cell line in which MK expression was the highest was transfeeted with different doses of MK siRNA. The expression of MK mRNA and protein was determined by real-time quantitative PCR and immunoflurescence staining. The cell adhesion was evaluated by MTF assay and invasion was examined by Boyden chamber method. Results Cell line MCF-7 expressed the highestlevel of MK mRNA in the 7 tested breast cancer cell lines. After being transfected with MK siRNA, MK mRNA and protein level of MCF-7 decreased in time- and dose- dependent manners. The adhesive and invasive ability of MCF-7 cell transfected with MK siRNA decreased in a dose dependent manner （P 〈 0. 01, P 〈 0.01 ）. Conclusions MK gene might play an important role in adhesion and invasion of human breast cancer ceils, siRNA transfection could effectively inhibit adhesion, migration, and invasion of human breast cancer cell.

关 键 词：乳腺癌 中期因子 RNA干扰 粘附 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]