摘要
目的研究甲基化转移酶抑制剂5-氮杂-2-脱氧胞苷(5-氮杂胞苷,AZA)对人肿瘤相关基因CHD5(染色质解旋酶DNA结合蛋白5)在结肠癌细胞中的基因转录的诱导作用以及其对细胞增殖的影响。方法 5μmol/L 5-氮杂胞苷分别作用于Lovo和SW480细胞,连续作用72 h后,用荧光定量PCR(qPCR)检测两种结肠癌细胞系中CHD5mRNA的表达,重亚硫酸盐测序(BSP)法分析启动子区的甲基化状况,MTT法分析5-氮杂胞苷对Lovo和SW480细胞增殖的影响。结果 5μmol/L 5-氮杂胞苷处理Lovo和SW480细胞72 h后,Lovo和SW480细胞中CHD5基因的发生了去甲基化,其mRNA重新表达,细胞生长受到抑制。结论 5-氮杂胞苷能够反转CHD5基因的甲基化状态,调控CHD5 mRNA表达,并能有效地抑制结肠癌细胞的增殖。
Objective To study the effect of 5-Aza-2-deoxycytidine （AZA） on re-expression of the hypermethylated and silenced CHD5 （ Chromodomain, helicase, DNA-binding 5 ） gene in human colon carcinoma cells and cell pro- liferation. Methods Lovo and SW480 cells were exposed to 5 μmoL/L AZA for 72 h, and then CHD5 expression was determined by quantitative RT-PCR （qPCR）, methylation status of CHD5 genes was determined by bisulfite sequencing PCR （ BSP）, and the proliferation of the cells was evaluated by MTF assay. Results After treatment with 5 μmol/L AZA, the promoter region of the CHD5 gene exhibited a demethylation status, and CHD5 mRNA was re-expressed. Meanwhile, the growth of Lovo and SW480 cells were reduced. Conclusions AZA may reverse methylation of CHD5 gene to regulate the expression of CHD5 gene, and effectively inhibit the ceil proliferation of human colon carcinoma cells.
出处
《基础医学与临床》
CSCD
北大核心
2013年第8期976-980,共5页
Basic and Clinical Medicine
基金
广东省自然科学基金(S2011040005521)
国家青年科学基金(81000226)
国家青年科学基金(81100383)
广东省医学科学技术研究基金(A2011360)
