5-aza-2’-deoxycitydine诱导胃癌细胞系TIMP3基因去甲基化和转录上调的实验性探讨  

Experimental Studies on 5-aza-2'-deoxycitydine Induction for Demethylation and Up-regulated Transcription of Human Gastric Cancer Cell Lines TIMP3 Gene

作　　者：关志宇[1] 戴冬秋[1] 孟春风[1] 

机构地区：[1]中国医科大学附属第一医院肿瘤外科,沈阳市110001

出　　处：《中国肿瘤临床》2006年第23期1334-1337,共4页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金(编号:30271477);教育部留学回国人员科研启动基金资助(编号:2002-247)

摘　　要：目的:研究人类胃癌细胞系中5-aza-2’-deoxycitydine诱导肿瘤抑制基因TIMP3mRNA和蛋白表达。方法:DNA甲基化抑制剂5-aza-2’-deoxycitydine处理人类胃癌细胞系SGC-7901。应用RT-PCR和Western-Blot方法检测TIMP3基因mRNA和蛋白表达。应用MSP分析TIMP3基因启动子甲基化状态。结果:5-aza-2’-deoxycitydine诱导TIMP3基因启动子去甲基化,上调TIMP3mRNA和蛋白表达。结论:甲基化异常是胃癌细胞TIMP3基因失活的原因之一,并可受去甲基化制剂调控表达。Objective： To investigate the effects of DNA methylation on cells. Methods： Gastric cancer cell lines （SGC-7901） was treated with DNA methyltransferase （DNMT） inhibitor, 5-aza- 2＇-deoxycytidine （5-aza-dC）. The expression of TIMP3 genes mRNA was detected using reverse transcription PCR （RT-PCR）. The proteinum expression was analyzedby the Western-Blot method. DNA methylation status of TIMP3 gene promoter was assayed by MSP （methylation-specific PCR）. Results： 5-aza-dC induced the demethylation of the TIMP3 gene promoter. The TIMP3 mRNA and proteinum expressions were obviously up-regulated by treatment with 5μmol/L for 24 hours. Conclusions： The hypermethylation of promoter region in CpG islands is a main mechanism of TIMP3 gene silencing in human gastric cancer cell, and may be controlled by demethylating agent 5-aza-dC.

关 键 词：胃癌 DNA甲基化 5-aza-2’-deoxycitydine基因表达 蛋白表达 

分 类 号：R735.2[医药卫生—肿瘤]