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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。
AIM: To investigate the effect of E-Cad on the proliferation of MC cells cultured on human gastric mucosa GES-1 immortalized cells induced by MNNG, and to explore the relationship between E-cad and Gli1 Gene relatedness. Methods: MTT colorimetric method was used to observe the growth cycle of GES-1 cells, MC cells and target cells. Semi-quantitative RT-PCR was used to observe the expression of Shh and Gli1 genes in three groups of cells. Fluorescence confocal microscope was used to observe the expression of E-Cad in three groups of cells. Results: The growth curve of MTT colorimetric assay showed that there was significant difference between MC group> target target interference group> GES-1 cell group, P <0.05. Semi-quantitative RT-PCR showed that the relative expression level of Gli-1mRNA in MC cells was 0.454 2 ± 0.008 4, which was higher than that in the target group (0.244 8 ± 0.002 6, t = 6.735, P = 0.015) The mean fluorescence intensities of E-Cad in GES-1 cells and target cells were 719 ± 114 and 278 ± 43, respectively, which were significantly higher than those in MC cells (64 ± 14, t = 9.437 and 4.357, P Values were 0.001 and 0.002 respectively. CONCLUSION: Gli1 may be involved in the regulation of E-Cad expression and decrease its expression in MC cells and enhance the in vitro proliferation activity of MC cells.