摘要
目的研究体内核转录因子KLF4是否与血管内皮生长因子(VEGF)启动子上游DNA序列相互作用。方法采用染色质免疫沉淀法鉴定人胃癌AGS细胞内KLF4与VEGF启动子上游DNA序列的相互作用,通过检索VEGF基因序列,发现在VEGF基因转录起始位点上游存在3个KLF4结合位点,针对这3个KLF4结合位点核心DNA序列(CACCC)设计3对PCR引物,利用KLF4抗体进行染色质免疫沉淀,PCR扩增。结果针对VEGF上游启动序列3个潜在的KLF4结合位点设计的引物,以KLF4特异性结合的DNA片段为模板,能够扩增出目的条带,与阴性对照组比较差异有显著性意义(P<0.05)。结论体内KLF4可能与VEGF启动子上游DNA序列相互作用。
Objective To investigate the interaction between transcriptional factor KLF4 and upstream DNA sequence of VEGF promoter. Methods The chromatin immunoprecipitation （CHIP） assay was used to identify the interaction between the nuclear factor KLF4 and VEGF promoter in the AGS cells. The total sequence of VEGF promoter was found in the Gene Bank. The eore DNA sequence to which KLF4 could bind was CACCC. It was found there were 3 CACCC responsive elements at upstream of the VEGF transcription initiation site in its promoter region. Three proper primer pairs were designed to amplify the fragments including the CACCC elements. The sequences to which KLF4 binding were chromatin immunoprecipitated by the anti-KLF4 antibody. Poiymerase chain reaction （PCR） was done using the sequences and the three primer pairs. Results The objective fragments were amplified by using the 3 designed primer pairs. As compared with the control group, the gray scale of the fragments in experimental group fragments showed significant difference （P〈0.05）. Conclusion In the human ceils, the nuclear factor KLF4 may bind to the upstream DNA sequences in the VEGF promoter region.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2008年第6期808-811,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.30672066)
