稳定表达LRIG1基因的人膀胱癌细胞株BIU87-LRIG1的建立和鉴定  

Establishment and identification of human bladder cancer cell line BIU87-LRIG1 stably expressing exogenous LRIG1 gene

作　　者：严泽军[1] 程跃[1] 杨为民[2] 叶章群[2] 

机构地区：[1]宁波大学医学院附属宁波市第一医院泌尿外科,315000 [2]华中科技大学同济医学院附属同济医院泌尿外科

出　　处：《中华实验外科杂志》2009年第10期1253-1254,共2页Chinese Journal of Experimental Surgery

摘　　要：目的建立稳定表达外源性LRIG1基因的人膀胱癌细胞株BIU87-LRIG1。方法利用脂质体介导的基因转染技术将质粒转染人人膀胱癌细胞株BIU87，实验分为3组：BIU87-LRIG1组（转染pLRIG1-GFP）、BIU87组（未转染质粒）和BIU87-neo组（转染pEGFP—N1）。经用G418筛选3周后出现抗性克隆，对其扩大培养后，应用逆转录-聚合酶链反应（RT—PCR）和Western blot分析各组中LIRG1 mRNA及其蛋白表达。结果RT—PCR结果表明，BIU87-LRIG1组中LRIG1 mRNA的表达水平（2．352±0．133）较BIU87-neo组（0．642±0．091）和BIU87组（0．516±0．045）明显升高；Western blot结果显示，BIU87-LRIG1组LRIG1蛋白表达水平（2．120±0．113）较BIU87组（0．946±0．075）和BIU87-neo组（1．132±0．076）明显升高，差异均有统计学意义（P〈0．01）。结论成功建立了稳定表达外源性LRIG1基因的人膀胱癌细胞株BIU87-LRIG1。Objective To establish human bladder cancer cell line BIU87-LRIG1 stably expressing exogenous LRIG1 gene. Methods Plasmids were transfected into BIU87 cells by Lipofectamine2000 reagent. The experiments were divided into 3 groups： BIU87-LRIG1 （transfected with pLRIG1-GFP）, BIU87 （no transfection）, and BIU87-neo （transfected with pEGFP-N1 ）. After transfection,the cells were selected by G418. Three weeks later, the resistant clones were chosen, and expanded. RT-PCR and Western blot were used to detect the expression of LRIG1 gene. Results RT-PCR analysis showed that LRIG1 mRNA level （2. 352 ±0. 133） in BIU87-LRIG1 group was significantly higher than that in BIU87 group （0. 516 ± 0.045 ） and BIU87-neo group （0.642 ± 0. 091 ）. Western blot analysis revealed that LRIG1 protein level in BIU87-LRIG1 group （2. 120 ± 0. 113） was significantly higher than that in BIU87 group （0.946± 0. 075 ） and BIU87-neo group （ 1. 132±0. 076） with the differences being statistically significant （P 〈 0. 01 ）. Conclusion Human bladder cancer cell line BIU87-LRIG1 was established successfully,which could stably express exogenous LRIG1 gene.

关 键 词：膀胱癌 转染 基因表达 鉴定 

分 类 号：R737.14[医药卫生—肿瘤] R331.141[医药卫生—临床医学]