甲基化寡核苷酸对SW480结肠癌细胞CHFR基因的诱导灭活作用及其临床意义  

Transfection of methylated oligonucleotides inactivates the CHFR gene in human colon cancer cell line SW480

作　　者：邱庆安[1] 吕云福[1] 陈一明[1] 林友刚[1] 伍海鹰[1] 刘宁[1] 

机构地区：[1]海南省人民医院普通外科,海南省海口市570311

出　　处：《世界华人消化杂志》2010年第33期3565-3568,共4页World Chinese Journal of Digestology

摘　　要：目的:探讨甲基化寡核苷酸诱导灭活SW480结肠癌细胞CHFR基因甲基化和表达情况对生物学行为的影响.方法:将与CHFR基因启动子序列互补的甲基化寡核苷酸转染进SW480结肠癌细胞内,分别应用MSP和RT-PCR检测转染前后CHFR基因启动子甲基化状态和mRNA表达.应用MTT法检测转染前后细胞增殖状态.结果:正常未转染寡核苷酸组和对照组SW480结肠癌细胞CHFR基因启动子表现未甲基化状态并表达CHFR基因mRNA.转染互补甲基化寡核苷酸后,CHFR基因启动子被诱导成甲基化状态,mRNA表达受到抑制而缺失,细胞增殖速度明显高于正常未转染寡核苷酸组和对照组;正常未转染寡核苷酸组和对照组细胞增殖速度无明显差异.结论:互补甲基化寡核苷酸可诱导SW480结肠癌细胞CHFR基因启动子甲基化状态并抑制其mRNA表达,促进细胞增殖.AIM： To investigate whether transfection of methylated oligonucleotides that are comple- mentary to the CHFR （checkpoint with forkhead and ring finger domains） gene promoter inacti- vates the CHFR gene in human colon cancer cell line SW480 and to analyze the effect of CHFR inactivation on the biological behavior of SW480 cells. METHODS： After methylated oligonucleotides that are complementary to the CHFR gene promoter were transfected into SW480 cells, MSP and RT-PCR were employed to detect the promoter methylation status and mRNA expression of the CHFR gene, respectively. MTT assay was used to detect the proliferation of SW480 ceils before and after transfection. RESULTS： In control SW480 cells, the promoterregion of the CHFR gene was unmethylated and CHFR mRNA expression could be detected. After transfection of methylated oligonucleotides, CHFR gene promoter methylation was induced and CHFR mRNA expression was suppressed. The proliferation of SW480 cells transfected with methylated oligonucleotides was significantly higher than that of control cells. CONCLUSION： Methylated oligonucleotides that are complementary to the CHFR gene promoter can induce CHFR gene promoter methylation, suppress CHFR mRNA expression, and promote cell proliferation in human colon cancer cell line SW480.

关 键 词：甲基化寡核苷酸 SW480结肠癌细胞 含叉头和环指域检控点基因 启动子 

分 类 号：R735.3[医药卫生—肿瘤]