E钙黏蛋白对MNNG诱导GES-1增殖影响及其与Gli1基因相关性研究

E-Cadherin expression in MNNG induced GES-1 with proliferation and its relationship with the expression of Gli1

目的:研究E钙黏蛋白(E-Cad)在甲基硝基亚硝基胍(MNNG)作用于人胃黏膜GES-1永生化细胞生成MC细胞后对增殖能力的影响,并探讨其与Gli1基因的相关性。方法:MTT比色法观察GES-1细胞组、MC细胞组及环靶明干扰细胞组的生长周期表达。半定量RT-PCR观察3组细胞Shh和Gli1基因的表达。荧光共聚焦观察3组细胞E-Cad的表达。结果:MTT比色实验绘制生长曲线显示,MC组>环靶明干扰组>GES-1细胞组,差异有统计学意义,P<0.05。半定量RT-PCR显示,MC细胞组的Gli-1mRNA相对表达水平为0.454 2±0.008 4,高于环靶明干扰组的0.244 8±0.002 6,t=6.735,P=0.015;荧光免疫显示,GES-1细胞组与环靶明干扰细胞组E-Cad的平均荧光强度分别为719±114与278±43,均明显高于MC细胞组的64±14,t值分别为9.437和4.357,P值分别为0.001和0.002。结论:Gli1可能参与调控E-Cad的表达,使其在MC细胞的表达减少,增强MC细胞的体外增殖活性。

OBJECTIVE：To explore E-Cadherin（E-Cad）expression in methylation nitroiminotetrahydro of nitroguanidine（MNNG）induced GES-1to MC cells and its relationship with the expression of Gli1.METHODS：The MTT was used to observe GES-1cells grew,MC cell grew and Cyclopamine interference group＇s cell growth cycle.Fluorescence confocal microscopy and Semi-quantitative RT-PCR were used to observe expression of E-Cadherin,ShhmDNA and Gli1mDNA.RESULTS：MTT colorimetric experimental growth curve displayed MC cell grewMC cellsGES-1cells grew（P0.05）.Semi-quantitative RT-PCR showed that the MC cell＇s Gli1mRNA was 0.454 2±0.008 4,it was higher than that of Cyclopamine interference group＇s 0.244 8±0.002 6（t=6.735,P=0.015）.The fluorescence immunoassay showed that GES-1cells and Cyclopamine interference group＇s mean fluorescence intensity was 719±114and 278±43,significantly higher than that of the MC cells group＇s 64±14（t=9.437,P=0.001;t=4.357,P=0.002）.CONCLUSION：Gli1may be involved in the regulation E-Cadherin expression by decreasing the expression in MC cells to strengthen the proliferative activity in MC cells.