胃泌素及其受体拮抗剂作用下胃癌细胞DNMT-1与MMP-2E-Cadherin甲基化相关性研究

摘要

Objective:To study gastrin and its antagonist on the expression of DNMT-1 and MMP-2E-Cadherin methylation in gastric cancer line MKN45 and evaluate their molecules relationship. Method:Separate gastrin and its antagonist’s effect on gastric cancer cells were divided into four groups:controlled group,gastrin group,proglumid group,and mixed group. SYBR GreenⅠQ-PCR method was used to detect expressions of DNMT-1 in all the cases. Genomic DNA modified by sodium bisulfite was used as a template. RQ-MSP detected the methylation status of the gene promoter of MMP-2E-cadherin. Result:DNMT-1 expression in gastrin group increased significantly in comparison to those of the controlled group(P<0.05). However,DNMT-1 expression in proglumid group decreased significantly in comparison to those of controlled group(P<0.05). There was a significant positive correlation between the expression of DNMT-1 and E-Cadherin methylation,but no correlation between MMP-2 methylation and the expression of DNMT-1. Conclusion:Gastrin can promote DNMT-1 expression. The expression of DNMT-1 may directly influence E-Cadherin methylation,yet may indirectly influence the expression pattern of MMP-2.%目的：通过检测胃泌素及丙谷胺干预下胃癌MKN45细胞DNMT-1基因表达与E-Cadherin和MMP-2基因甲基化变化的相关性。方法：胃癌细胞株分别在胃泌素和丙谷胺干预下分为空白组、胃泌素组、丙谷胺组和混合组，应用SYBR Green I Q-PCR检测各组DNMT-1基因表达水平；运用RQ-MSP法检测用亚硫酸氢盐修饰后的基因组DNA为模板的各组基因甲基化状态。结果：胃泌素组与空白组相比， DNMT-1表达明显增高，而丙谷胺组则表达明显减低（P<0.05）；DNMT-1表达量与E-Cadherin甲基化变化存在明显相关性，而与MMP-2甲基化并无明显相关性。结论：胃泌素可促进DNMT-1表达，DNMT-1可能直接影响E-Cadherin甲基化，而其可能间接影响MMP-2表达模式。