NDRG-1异常甲基化及对胃癌细胞恶性生物学行为影响的研究

目的:探讨N-myc下游调节基因1(NDRG-1)甲基化对胃癌细胞恶行生物学行为的影响。方法:用免疫组织化学法检测NDRG-1在胃癌组织及癌旁组织中的表达;qRT-PCR与Western blot检测胃癌细胞SGC7901及胃正常黏膜上皮细胞RGM-1中NDRG-1的表达,MSP法检测胃癌组织及癌旁组织、胃癌细胞及正常胃上皮黏膜细胞中NDRG-1的甲基化状态。MSP法检测5-Aza-CdR处理的SGC7901细胞的甲基化水平;应用qRT-PCR、Western blot及免疫荧光染色与Transwell、划痕闭合实验和流式细胞术检分别检测5-Aza-CdR处理的SGC7901及si-NDRG-1转染5-Aza-CdR处理的SGC7901细胞中NDRG-1表达水平与细胞侵袭、迁移及凋亡能力变化。结果:胃癌组织及细胞中NDRG-1低表达,且NDRG-1甲基化。与对照组相比,5-Aza-CdR组SGC7901细胞NDRG-1甲基化消除,NDRG-1表达升高,侵袭细胞数与划痕闭合率降低,细胞凋亡能力升高。与5-Aza-CdR+si-NC组比较,5-Aza-CdR+si-NDRG-1组SGC7901细胞NDRG-1表达降低,侵袭细胞数与划痕闭合率升高,细胞凋亡能力降低。结论:胃癌细胞NDRG-1甲基化可促进细胞增殖,抑制细胞凋亡。

Abnormal methylation of N-myc downstream regulated 1 in gastric cancer and its influence on the malignant biological behavior of gastric cancer cells

Objective:To investigate the effect of N-myc downstream regulated 1(NDRG-1)methylation on the biological behavior of gastric cancer cells.Methods:Immunohistochemistry was used to detect the expression of NDRG-1 in gastric cancer tissues and adjacent tissues;qRT-PCR and Western blot were used to detect the expression of NDRG-1 in gastric cancer cells SGC7901 and normal gastric mucosal epithelial cells RGM-1;MSP method was used to detect the methylation status of NDRG-1 in gastric cancer tissues and adjacent tissues,gastric cancer cells and normal gastric epithelial mucosal cells.MSP method was performed to detect the methylation level of SGC7901 cells treated with 5-Aza-CdR,qRT-PCR,Western blot,immunofluorescence staining,Transwell,scratch closure test and flow cytometry were used to detect the expression of NDRG-1,the ability of cell invasion,migration and apoptosis in 5-Aza-CdR-treated SGC7901 cells and si-NDRG-1 transfected 5-Aza-CdR-treated SGC7901 cells.Results:NDRG-1 was down-regulated and methylated in gastric cancer tissues and cells.Compared with the Control group,the NDRG-1 methylation status of SGC7901 cells was eliminated,the NDRG-1 expression was increased,the number of invaded cells and scratch closure rate were decreased,and the apoptosis ability was increased in the 5-Aza-CdR group.Compared with 5-Aza-CdR+si-NC group,the expression of NDRG-1 in SGC7901 cells was decreased,the number of invaded cells and scratch closure rate were increased,the ability of apoptosis was decreased in 5-Aza-CdR+si-NDRG-1 group.Conclusion:Methylation of NDRG-1 in gastric cancer cells could promote cell proliferation and inhibit cell apoptosis.