染色质免疫沉淀技术分析肺腺癌A549细胞中p53蛋白与p21^(CIP1)和Bim基因转录启动子的结合

Detection of p53 Binding to p21^(CIP1) and Bim Gene Promoters by Chromatin Immunoprecipitation

为了检测肺腺癌A549细胞内抑癌蛋白p53与CDK抑制蛋白p21CIP1和Bim基因转录调控区的结合情况,采用染色质免疫沉淀技术,用p53特异性抗体沉淀DNA,PCR检测p21CIP1和Bim基因5′端特异性序列。结果表明,在抗体免疫沉淀的DNA片段中扩增出p21CIP1和Bim基因5′端的特异性序列。因此证实在A549细胞内,p53蛋白可与p21CIP1和Bim基因转录启动子的特异区域结合,进而参与两基因的表达调控。

It was to detect and verify p53 protein binding to p21^CIPI and Bim gene promoter regions in A549 cells in vivo,by using chromatin immunoprecipitation （CHIP） assay and specific p53 antibody involved in. The immunoprecipitated DNA fragments were amplified through PCR using the specific p21^CIPI and Bim primers. Results indicated that p21^CIPI and Bim gene specific fragments were detected in the DNA fragments immunoprecipitated by p53 antibody. Thus it proved that p53 protein could bind to the promoter regions of p21^CIPI and Bim genes in A549 ceils and involved in upregulation of these two gene expression.