盐酸普鲁卡因对人结肠癌HT-29细胞Syk基因甲基化及表达的影响  

The Effect of Procaine Hydrochloride in Methylation Status and Expression Level of Syk Gene in Human Colon Cancer HT-29 Cells

作　　者：高曰文[1] 朱耀明[2] 王艳林[3] 朱晨宇[2] 秦烨[3] 胡汉卿[2] 韩钰[3] 

机构地区：[1]三峡大学医学院,宜昌443002 [2]三峡大学第一临床医学院,宜昌443002 [3]三峡大学分子生物所,宜昌443002

出　　处：《生物技术通报》2010年第12期173-177,共5页Biotechnology Bulletin

基　　金：三峡大学研究生科研创新基金项目(2009-35)

摘　　要：旨在探讨盐酸普鲁卡因(procaine,PCA)对人结肠癌HT-29细胞Syk基因甲基化及表达的影响。应用巢式双重甲基化特异性聚合酶链反应(Methylation-specific PCR,MSP)检测盐酸普鲁卡因处理前后HT-29细胞中Syk基因启动子的甲基化水平。逆转录-聚合酶链反应(RT-PCR)和蛋白印迹技术观察HT-29细胞内Syk基因表达情况。MSP检测发现人结肠癌HT-29细胞中Syk基因存在甲基化,经盐酸普鲁卡因处理能够使Syk基因甲基化水平下降。RT-PCR和蛋白印迹分析结果显示,人结肠癌HT-29细胞经盐酸普鲁卡因处理后Syk基因表达上调。人结肠癌HT-29细胞中Syk基因启动子甲基化导致基因表达沉默,盐酸普鲁卡因能逆转Syk基因启动子区域CpG岛甲基化,使Syk基因活化并表达上调,提示盐酸普鲁卡因具有治疗结肠癌的潜在应用价值。This study was to investigate the effect of procaine hydrochloride in methylation status and expression level of Syk gene in human colon cancer HT-29 cells.The nest-MSP was used to detect the methylation status of Syk gene in HT-29 cells before and after treatment by procaine hydrochloride.Reverse transcription polymerase chain reaction（RT-PCR）and Western blotting assay were used to determine expression levels of Syk gene.Results showed that hypermethylation in the promoter of Syk gene in human colon cancer HT-29 cells was observed by MSP.Treating HT-29 with procaine hydrochloride resulted in demethylation of Syk promoter clearly.In the same time,RT-PCR and Western blotting assay showed that Syk gene has an up regulation after treatment by procaine hydrochloride in human colon cancer HT-29 cells.The methylation of Syk gene promoter can lead to the loss of gene expression in human colon carcinoma HT-29 cells,procaine hydrochloride can reverse the methylation status of Syk gene promoter and result in an reactivation and up regulation of Syk gene.Thus,procaine is a promising candidate agent for therapy of human colon cancer.

关 键 词：盐酸普鲁卡因 SYK基因 甲基化 

分 类 号：R735.35[医药卫生—肿瘤]