MNNG、CDCA构建胃黏膜上皮细胞GES-1肠化生模型的可行性

摘要

目的 探讨N-甲基-N′-硝基-N-亚硝基胍(MNNG)、鹅脱氧胆酸(CDCA)构建胃黏膜上皮细胞GES-1肠化生模型的可行性.方法 体外培养GES-1细胞,分别给予不同浓度CDCA、MNNG处理24 h,采用CCK-8法检测450 nm波长处各孔光密度( OD)值,确定CDCA、MNNG对GES-1 细胞具有生长抑制作用,二者的10%抑制浓度(IC10)分别为0.14、51.89 μmol/L.另取GES-1细胞随机分为CDCA组、MNNG组,分别加入IC10的CDCA、MNNG,同时设未加药物处理的细胞为对照组,培养24 h时采用实时荧光定量PCR法检测黏蛋白1(MUC1)mRNA、黏蛋白2(MUC2) mRNA及尾型同源框转录因子2(CDX2)mRNA相对表达量.结果 与对照组比较,CDCA组、MNNG组MUC1 mRNA相对表达量均降低,MNNG组MUC2 mRNA相对表达量降低,组间比较P均＜0.05.三组间CDX2 mRNA相对表达量比较P＞0.05.结论 CDCA、MNNG可抑制胃黏膜上皮细胞生长,可用于构建胃黏膜上皮细胞的肠化生模型;下调胃特异性基因MUC1表达、上调肠特异性基因CDX2表达可能是二者的作用机制.%Objective To investigate the feasibility of using N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and goose deoxycholic acid (CDCA) to construct intestinal metaplasia (IM) model of gastric epithelial cells GES-1.Methods GES-1 cells were cultured in vitro, and then were treated with different concentrations of CDCA and MNNG for 24 h, and the values of OD at 450 nm wavelength were detected by CCK-8.The results showed that CDCA and MNNG had a growth inhibitory role on GES-1 cells, with 10%inhibition concentration (IC10) of 0.14 and 51.89 μmol/L, respectively.GES-1 cells were ran-domly divided into the CDCA group and MNNG group , and then were treated with CDCA and MNNG of IC 10, respectively.At the same time, GES-1 cells without medicine were taken as the control group.At 24 h, the expression of MUC1, MUC2, and CDX2 mRNA were detected by RT-PCR.Results Compared with the control group , the relative expression of MUC1 mRNA in the CDCA group and MNNG group decreased , the relative expression of MUC2 mRNA in the MNNG group decreased , and significant difference was found between every two groups (all P＜0.05).Conclusions CDCA and MNNG can inhibit the growth of gastric mucosa epithelial cells.Therefore, both CDCA and MNNG can be used to build IM models of gastric muco-sal cells.