HepaCAM联合吡柔比星抑制膀胱癌BIU-87细胞的增殖  

Combined treatment of hepa CAM and pirarubicin inhibits the proliferation of bladder cancer BIU-87 cells

作　　者：吕长坤[1] 万雪莲[2] 马菲菲[2] 杜红飞[1] 杨雪[1] 罗春丽[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016 [2]商丘医学高等专科学校医学检验技术教研室,商丘476100

出　　处：《重庆医科大学学报》2015年第2期234-238,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:81072086)

摘　　要：目的:研究肝细胞黏附分子(hepatocyte cell adhesion molecule,hepa CAM)联合吡柔比星(pirarubicin,THP)对膀胱癌细胞BIU-87的增殖影响。方法:将BIU-87细胞分为空白处理组、hepa CAM过表达腺病毒(adenovirus-GFP-hepa CAM,Ad-GFPhepa CAM)处理组、THP处理组、Ad-GFP-hepa CAM联合THP处理组。噻唑蓝(methyl thiazolyl tetrazolium,MTT)检测细胞增殖,流式细胞仪(flow cytometry,FCM)检测细胞周期,real-time PCR检测细胞周期相关基因cyclin D1,CDK2的m RNA表达。将空白、空载腺病毒(adenovirus-GFP,Ad-GFP)、hepa CMA过表达腺病毒(Ad-GFP-hepa CAM)处理的BIU-87细胞分别注射于裸鼠皮下,观察裸鼠成瘤情况并记录瘤体体积,4周后处死裸鼠。切取各组瘤体组织,组织切片行HE染色和免疫组化检测hepa CAM蛋白的表达。结果:Ad-GFP-hepa CAM联合THP处理组,与THP处理组比较,BIU-87细胞增殖受到明显抑制(F=2.702,P=0.00);细胞周期明显阻滞于G0/G1期(F=10.989,P=0.000);cyclin D1、CDK2的m RNA表达明显下调(F=5.4299,P=0.000)。Ad-GFPhepa CAM处理组的BIU-87细胞成瘤体积明显低于空白处理组(F=12.587,P=0.000)。结论:hepa CAM基因能够增强吡柔比星对膀胱癌BIU-87细胞的增殖抑制作用。Objective：to investigate the effects of combination treatment of hepa CAM and pirarubicin（THP）on the proliferation of BIU-87 cells. Methods：BIU-87 cells were designed as blank treatment group,adenovirus-GFP-hepa CAM treatment group（Ad-GFPhepa CAM treatment group）,THP treatment group,Ad-GFP-hepa CAM and THP combination treatment group. MTT was used to detect cell proliferation;cell cycle was evaluated by flow cytometry;the m RNA expressions of cyclin D1 and CDK2 were examined by real-time PCR. Cells that were designed as blank treatment group,Ad-GFP treatment group,Ad-GFP-hepa CAM treatment group were subcutaneously injected in nude mice. The volume of nude mice was evaluated. After 4 weeks,all mice were sacrificed,transplanted tumors were excised and tumor tissues were used to perform HE staining and immunohistochemistry was performed to examine hepa CAM expression. Results：Ad-GFP-hepa CAM combined with THP exhibited more intense inhibitory effects on proliferation of BIU-87 cells than THP alone（F=2.702,P=0.000）. Meanwhile,the cell in G0/G1 was significantly increased（F=10.989,P=0.000）and m RNA levels of cyclin D1 and CDK2 were significantly lowered in Ad-GFP-hepa CAM and THP combination treatment group than in THP group（F=5.4299,P=0.000）. In addition,the volume of tumors induced by hepa CAM-expression BIU-87 cells was significantly decreased compared with that of blank treatment group（F=12.587,P=0.000）. Conclusion：Hepa CAM could enhance the inhibitory effects of THP on the proliferation of bladder cancer BIU-87 cells.

关 键 词：HEPA CAM 吡柔比星 膀胱癌 CYCLIN D1 CDK2 

分 类 号：R73[医药卫生—肿瘤]