吡格列酮对膀胱正常移行上皮细胞及膀胱癌细胞生长的影响

Effect of pioglitazone on the growth of normal bladder transitional epithelial cells and bladder cancer cells

以不同浓度吡格列酮处理膀胱正常移行上皮细胞及膀胱癌J82细胞24h、48h、72h、5d、8d和10d．观察细胞生长状态，MTT检测细胞生长抑制率，流式细胞仪分析细胞凋亡，实时定量PCR和Western印迹法检测周期素D1、p53、Bcl-2、Bax的mRNA和蛋白表达。结果显示，10μmol／L吡格列酮处理膀胱正常细胞24、48、72h后细胞生长明显受抑制（均P〈0．05），细胞凋亡增加[24h（18．8±2．1对9．4±1．7）％、48h（29．9±1．3对10．7±1．1）、72h（49．7±2．3对11．3±1．3）％．P〈0．05]．J82细胞未见明显影响．10μmol／L吡格列酮处理2种细胞72h内周期素D1、p53、Bcl-2、Bax的mRNA和蛋白表达均无明显变化，作用8d后J82细胞周期素D1、p53蛋白表达降低，提示吡格列酮不增加膀胱正常移行上皮细胞癌变趋势。

The normal bladder transitional cells and bladder cancer J82 cells were treated with various concentrations＇of pioglitazone for 24 h, 48 h, 72 h, 8 d, and 10 d. The growth of the cells was tested by MTF. Apoptosis rate was detected by flow cytometry technology. Realtime-PCR and Western blot were used to analyze the mRNA and protein expressions of p53, cyclin DI, Bcl-2, and Bax. The results showed that the cell viability of normal bladder cells was decreased 24, 48, and 72 h after 10 μmol/L pioglitazone treatment （ all P〈0.05 ） while the cell apoptosis was increased [ 24 h （ 18.8 ± 2.1 vs 9.4 ±1.7 ） % , 48 h （ 29.9 ±1.3 vs 10.7 ±1.1 ）, 72 h （49.7 ±2.3 vs 11.3± 1.3 ）% , P〈0.05 1. Pioglitazone had no effect on the growth and proliferation of J82 cells. There were no significant differences in mRNA and protein expressions of p53, cyclin D1, Bcl-2, and Bax within 72 h after pioglitazone treatment in this two kinds of cells. But, the protein expressions of cyclin D1 and p53 in J82 cells were decreased after pioglitazone treatment for 8 days. These results suggest that pioglitazone does not increase the risk of bladder cancer in normal transitional cells.