目的:研究抑癌候选基因NDRG2在体外对人食管鳞癌细胞系Eca-109增殖和凋亡的影响。方法:腺病毒介导NDRG2转染人食管鳞癌细胞系Eca-109,逆转录酶-多聚酶链反应(RT-PCR)和Western blot法分别检测NDRG2基因的mRNA和蛋白表达水平,甲基噻唑基四唑法(MTT法)绘制细胞生长曲线,流式细胞仪(FCM)分别分析细胞周期和细胞凋亡。结果:腺病毒介导NDRG2转染人食管鳞癌细胞系Eca-109后,NDRG2的mRNA和蛋白表达水平明显上调,细胞增殖受到明显抑制(P〈0.05);流式细胞仪检测显示,与对照组相比,感染后48小时Eca-109细胞G2期细胞明显减少(P〈0.01),S期细胞明显增多(P〈0.01);转染后的Eca-109细胞中凋亡细胞明显增多,感染48h后达16.0%。结论:过表达NDRG2可明显抑制人食管鳞癌细胞Eca-109的增殖并诱导细胞凋亡。
Objective:To investigate the function of candidate tumor suppressor gene NDRG2 on proliferation and apoptosis of human esophageal cancer cell line Eca-109.Methods:Eca-109 cells were transfected with adenovirus vector containing NDRG2 gene. The expression of message RNA and protein of NDRG2 were detected by RT-PCR and Western blot respectively. The cell growth curve was drawn by methyl thiazolyl tetrazolium assay(MTT). Flow cytometry (FCM) was adopted to analyze quantitatively the cell cycle and apoptosis.Results:NDRG2 expression was significantly increased in both mRNA and protein levels in Eca-109 after transfecting with adenovirus vector carrying NDRG2 gene. As compared with normal Eca-109 cells, the growth was markedly slowed down in NDRG2 over expressing cells. The G2 phase cells were decreased significantly (P〈0.01), whereas S phase cells were obviously increased. Moreover, apoptotic cells which presenting typical morphologic changes of cell apoptosis, reached 16% after adenovirus transfection.Conclusion:NDRG2 gene can significantly inhibit cell growth and markedly induce the apoptosis of Eca-109 cells in vitro.