N—MYC下游调节基因-2对膀胱癌细胞T24增殖的抑制作用

Inhibitory effects of N-MYC down-stream regulated gene-2 on proliferation of bladder carcinoma cell line T24 in vitro

目的观察抑癌基因N—MYC下游调节基因-2（NDRG2）在不同膀胱癌细胞株的表达水平及对T24细胞体外增殖的抑制作用。方法采用Westernblot及逆转录-聚合酶链反应（RT—PCR）检测3种膀胱癌细胞及正常上皮组织细胞的NDRG2表达，使用pAD—CMV腺病毒感染的方法观察NDRG2对T24细胞的作用，乳糖操纵子z（Lacz）为阳性对照，并设空白对照，流式细胞仪（FCM）连续检测细胞的周期及凋亡。结果3种膀胱癌细胞株的NDRG2表达水平相对较低，其中T24细胞表达水平最低，FCM检测显示，感染组同阳性对照组及空白组比较，细胞G，期阻滞[3组分别为（56．26±3．02）％、（50．394-3．13）％和（47．414-2．63）％]，细胞凋亡增加[3组分别为（12．06±1．33）％、（4．27±1．01）％和（2．454-0．86）％]。结论NDRG2基因可能参与了膀胱癌的发病，腺病毒介导的人NDRG2基因可抑制T24细胞的增殖。

Objective To detect the expression level of N-MYC down-stream regulated gene-2 （NDRG2） in different bladder carcinoma cell lines, and investigate the inhibitory effects of NDRG2 on proliferation of T24 cells mediated by adenovirus in vitro. Methods The expression of NDRG2 protein and mRNA was detected by using Western blotting and reverse transcription-polymerase chain reaction （ RT- PCR） respectively in three bladder cancer cell lines. PAD-NDRG2 adenoviruses were used to infect the T24 cells. The changes in cell cycle and apoptosis were detected by using flow cytometry （FCM） at the 72nd h after the infection. Results The results of Western blotting and RT-PCR revealed that the three bladder carcinoma cell lines had relatively lower NDRG2 protein and mRNA expression levels. T24 cells had the lowest level among the three cell lines. As compared with the positive control and the blank con- trol, pAD-CMV-NDRG2-infected cells showed an increase in G1 phase at 72nd h [ （56. 26 ± 3.02） % , （50. 39 ±3.13）% and （47.41 ±2. 63）% respectively]. Furthermore, NDRG2-induced apoptosis was al- so increased at 72nd h in comparison with the controls [ （ 12. 06 ± 1.33 ） %, （4. 27 ± 1.01 ） % and （2. 45 ± 0. 86% ） % ]. Conclusion The NDRG2 might be involved in the pathogenesis of bladder carcinoma. Adenovirns-mediated NDRG2 could suppress the proliferation of T24 cells significantly.