河南农业科学Issue(11)：126-131,6.
5 Aza dC对奶牛乳腺上皮细胞PPARγ表达的影响
The Effect of 5-Aza-2′-Deoxycytidine on the Expression of PPARγin Dairy Cow Mammary Epithelial Cells in vitro
摘要
Abstract
This experiment was to explore the influences of 5-aza 2′-deoxycytidine(5-Aza-dC)on the methylation status of promoter region of PPARγgene and its expression in dairy cow mammary epithelial cells(DCMECs).DCMECs were cultured invitro and treated with 0.1,0.5,1.0,5.0μmol/L 5-Aza-dC,methyltransferase activity was measured by EpiQuikTM DNA methyltransferase assay kit;CASY○R-technology was applied to measure cell viability and proliferation;bisulphite genomic sequencing PCR(BSP)technique was applied to detect the methylation state of the promoter region of PPARγgene in DCMECs;qRT-PCR was applied to detect PPARγmRNA expression;the protein expression level of PPARγwas determined by Western blot.Results showed that 0.1μmol/L 5-Aza-dC had no significante effect on cell viability and proliferation of DCMECs,and methyltransferase activity was significantly decreased at 9 6 h;compared with control group,methylation level of promoter region of PPARγgene in DCMECs was partially reduced after treating with 5-Aza-dC,and the mRNA and protein expression of PPARγwere increased significantly.Our results suggests that 5-Aza-dC can reduce methylation level of PPARγpromoter region,and promote the mRNA and protein expre s sion of PPARγgene in DCMECs.关键词
5 氮杂 2′-脱氧胞苷/奶牛乳腺上皮细胞/PPARγ基因/DNA甲基化Key words
5-aza-2′-deoxycytidine/daily cows mammary epithelial cells/PPARγ gene/methylation分类
农业科技引用本文复制引用
边艳杰,王杰,王春梅,李庆章..5 Aza dC对奶牛乳腺上皮细胞PPARγ表达的影响[J].河南农业科学,2014,(11):126-131,6.基金项目
国家重点基础研究发展（973）计划项目 （）
