摘要
过氧化物酶体增殖因子活化受体δ基因(PPARδ)与结直肠癌相关,但其转录调控机制尚不清楚.本研究构建了PPARδ基因的启动子荧光素酶报告基因载体,用删除法定位了启动子活性区域,并通过核酸重叠法和电泳迁移率变更实验,将该区域缩小到30bp,预测该区域可能起作用的转录因子结合位点;通过启动子定点突变分析和核酸诱骗实验,发现真正起作用的是-156^-127区域内NF-κB的结合位点.电泳迁移率变更法和超迁移分析实验,证实NF-κB能够与该区域结合,并且在抑制NF-κB的DNA结合活性后,PPARδmRNA的表达明显降低.这些实验表明,在Lovo细胞中,转录因子NF-κB参与了PPARδ的表达调控.
Peroxisome proliferator-activated receptors gene（PPARδ） has been validated to be correlated with colorectal carcinogenesis; however, the mechanism of transcriptional regulation remains unclear. Our experiment is to address this issue. In this study, the PPARδpromoter was constructed into luciferase report gene vector. A functional region in the promoter was identified by deletion analysis and then the length was restricted to 30 bp by overlapping methods and EMSA. Possible transcriptional factor binding sites residing in the region were predicted, and site-directed mutagenesis analysis and decoy analysis demonstrated that the functional transcriptional factor binding site was the NF-κB binding site residing in a region extending from - 156 to - 127 in the PPARδ promoter. EMSA and supershift assay confirmed that NF-κB could bind to this region, and PPARδ mRNA expression decreased significantly after the DNA binding activity of NF-κB was inhibited. In conclusion, NF-κB is the transcriptional factor that contributes to the PPARδexpression in Lovo cells.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2008年第10期931-937,共7页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金资助项目(No.30471914)~~
关键词
过氧化物酶体增殖因子活化受体δ
NF-ΚB
结直肠癌
转录调控
peroxisome proliferator-activated receptorδ （PPARδ）PPARδ
NF-κB
colorectal carcinoma
transcriptional regulation
作者简介
联系人Tel：010—82805138，E-mail：zbmao@bjmu．edu．cn
