尿道致病性大肠杆菌黏附人尿路膀胱癌细胞的实验研究  

Study on the adhesion of uropathogenic Escherichia coli to human urinary tract bladder cancer cells

作　　者：谷超[1] 王海涛[2] 周晓冬[2] 侯敏[3] 李光[1] 

机构地区：[1]天津医科大学基础医学院生物学教研室,天津300070 [2]天津市泌尿外科研究所肿瘤免疫室 [3]天津市胸科医院检验科

出　　处：《山西医科大学学报》2015年第10期973-976,共4页Journal of Shanxi Medical University

基　　金：863国家高技术研究发展计划资助项目(2012AA021003);天津医科大学科学基金资助项目(2008KY04)

摘　　要：目的检测尿道致病性大肠杆菌(UPEC)菌株毒力基因,并将人膀胱癌细胞系EJ细胞应用于UPEC的黏附作用研究。方法利用PCR和复合PCR方法检测UPEC132菌株毒力基因pap C,hly和cnf1;制备EJ的单层细胞爬片用于UPEC和阴性对照菌株E.coli K-12p678-54的黏附试验;分别于不同作用时间观察黏附细胞形态学变化,并计算黏附率和黏附指数。结果PCR检测显示UPEC132菌株具有毒力基因pap C,hly和cnf1,将其作用于EJ细胞后,15 min细菌开始黏附,120 min达到高峰。而阴性对照组结果均为阴性。经光镜镜检,UPEC黏附EJ细胞后使其产生明显的形态学变化。结论 EJ细胞可用于UPEC的黏附试验,为建立UPEC细胞感染模型和深入探究UPEC的致病机制奠定基础。Objective To explore the adhesion of uropathogenic Escherichia coli（ UPEC） to human bladder cancer cell line EJ.Methods The virulence genes pap C,hly and cnf1 of UPEC132 strain were detected by PCR and multiplex PCR. EJ monolayer cell slides were prepared for the adhesion test of UPEC132 and negative control strain E. coli K-12p678-54. The morphological changes of adhered cells at different stages were observed,and the adhesion rates and indexes were calculated. Results The virulence genes pap C,hly and cnf1 were expressed in UPEC132 by PCR. Under the action of UPEC132 with pap C,hly and cnf1,the adhesion to EJ cells began at 15 min and reached the peak at 120 min. However,the results for E. coli K-12p678-54 were negative. UPEC adhesion generated obvious morphological changes in EJ cells under microscopy. Conclusion EJ cells can be used in UPEC adhesion test,which lay the foundation for the establishment of UPEC cellular infection model and the further research on its pathogenic mechanism.

关 键 词：尿道致病性大肠杆菌 膀胱癌 EJ细胞 黏附率 黏附指数 毒力基因 

分 类 号：R737.14[医药卫生—肿瘤]