促肝细胞再生磷酸酶-1基因对肺癌细胞侵袭的影响  

Effects of phosphatase of regenerating liver cell-1 on invasion of human lung cancer cells

作　　者：詹洪峰[1] 周永静[1] 范钰[1] 

机构地区：[1]江苏大学附属人民医院肿瘤研究所,镇江212002

出　　处：《中华实验外科杂志》2010年第8期1119-1121,共3页Chinese Journal of Experimental Surgery

基　　金：中国博士后科学基金资助项目（2003033547）

摘　　要：目的 探讨促肝细胞再生磷酸酶-1（PRL-1）基因对肺癌细胞侵袭的影响及其机制.方法 应用PRL-1基因小干扰RNA（siRNA）转染处理肺癌细胞株A549后,分别采用实时定量逆转录-聚合酶链反应（RT-PCR）和Western blot检测PRL-1基因和基质金属蛋白酶（MMP）-2、MMP-7和MMP-9 mRNA和蛋白水平,分别采用软琼脂集落培养实验和Boyden小室模型实验检测癌细胞的锚着不依赖性增殖和侵袭能力.结果 siRNA转染组PRL-1 mRNA和蛋白水平明显下调,且呈浓度和时间依赖性（P＜0.01）.软琼脂集落形成实验显示,3.125、6.25和12.5 nmol/L siRNA组集落形成数分别为17.8±1.6、13.6±1.5、8.8±1.4,而对照组为22.6±1.8（P＜0.05）;Boyden小室模型实验显示,3.125、6.25和12.5 nmol/L siRNA组穿过滤膜的细胞分别为33.6±2.1、19.5±1.9、8.1±1.8,而对照组为49.4±2.3（P＜0.05）.同时发现,转染组细胞MMP-2、MMP-7、MMP-9基因mRNA和蛋白水平明显下调.结论 PRL-1 siRNA转染可抑制肺癌细胞侵袭,其机制可能与下调MMP-2、MMP-7、MMP-9表达有关.Objective To study the effects of phosphatase of regenerating liver cell-1 （PRL-1） small interfering RNA （siRNA） on invasion of lung cancer cells and the mechanism. Methods After lung cancer cell line A549 cells were transfected by PRL-1 siRNA, the mRNA and protein expression levels of PRL-1 and matrix metalloproteinase-2 and-9 were detected by real time reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting, respectively. The anchorage-independent growth was examined using colon formation in soft agar, and invasion ability was evaluated by boyden chamber model. Results The anchorage-independent growth showed that the colonies were 17.8 ± 1.6, 13.6 ± 1.5, 8.8 ± 1.4, and 23.6 ±1.8 in different groups （3.125, 6.25, 12.5 nmol/L siRNA and control）, respectively. The boyden chamber results revealed that the number of invasion cells was 33.6 ± 3.1, 19.5 ±1.9, 8.1 ±1.8 and 49.4 ±3.3 in different groups （3.125, 6.25, 13.5 nmol/L siRNA, and control） , respectively. The mRNA and protein expression levels of MMP-2,-7 and-9 in transfection group with PRL-1 siRNA were down-regulated as compared with control group in lung cancer cells. Conclusion PRL-1 siRNA can inhibit invasion through down-regulating MMPs of lung cancer cells.

关 键 词：肺肿瘤 侵袭 基质金属蛋白酶 

分 类 号：R734.2[医药卫生—肿瘤]