HepaCAM联合吡柔比星抑制膀胱癌BIU-87细胞的增殖

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

摘要:

目的：研究肝细胞黏附分子（hepatocyte cell adhesion molecule，hepaCAM）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、hepaCAM过表达腺病毒（adenovirus-GFP-hepaCAM，Ad-GFP-hepaCAM）处理组、THP处理组、Ad-GFP-hepaCAM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cyclinD1，CDK2的mRNA表达。将空白、空载腺病毒（adenovirus-GFP，Ad-GFP）、hepaCMA过表达腺病毒（Ad-GFP-hepaCAM）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hepaCAM蛋白的表达。结果：Ad-GFP-hepaCAM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）；cyclinD1、CDK2的mRNA 表达明显下调（F=5.4299，P=0.000）。Ad-GFP-hepaCAM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：hepaCAM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。

Abstract:

Objective：to investigate the effects of combination treatment of hepaCAM and pirarubicin（THP） on the proliferation of BI－U-87 cells. Methods：BIU-87 cells were designed as blank treatment group，adenovirus-GFP-hepaCAM treatment group（Ad-GFP-hepaCAM treatment group），THP treatment group，Ad-GFP-hepaCAM and THP combination treatment group. MTT was used to detect cell proliferation；cell cycle was evaluated by flow cytometry；the mRNA expressions of cyclinD1 and CDK2 were examined by re－al-time PCR. Cells that were designed as blank treatment group，Ad-GFP treatment group，Ad-GFP-hepaCAM 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 hep－aCAM expression. Results：Ad-GFP-hepaCAM 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 mR－NA levels of cyclinD1 and CDK2 were significantly lowered in Ad-GFP-hepaCAM and THP combination treatment group than in THP group（F=5.4299，P=0.000）. In addition，the volume of tumors induced by hepaCAM-expression BIU-87 cells was significantly decreased compared with that of blank treatment group（F=12.587，P=0.000）. Conclusion：HepaCAM could enhance the inhibitory ef－fects of THP on the proliferation of bladder cancer BIU-87 cells.