CAJ | 학술논문

　　目的鉴定整合素β6基因在HepG2细胞中的主要转录调控区，为进一步研究αvβ6在肝癌发生发展过程中的转录调控机制奠定基础。方法利用β6基因转录起始位点附近1135 bp基因序列经PCR进行不同长度的缺失扩增，并连接到pGL2-basic载体，构建重组质粒，利用双荧光素酶报告基因分析系统检测启动子的转录活力，并利用Transcription Element Search System预测β6基因主要转录调控区潜在转录因子结合位点。结果获得4个含不同长度β6基因序列的重组质粒；当β6基因在距转录起始位点-458 bp截短到-187 bp时，转录活力降低约50％，在这个转录调控区存在YY1、AP-1和c-Myb等与肝癌相关的转录因子结合位点。结论在HepG2细胞中，β6基因启动子区的-458 bp/-187 bp区域是其主要的转录调控区，YY1、AP-1和c-Myb三个转录因子存在该区域，并可能是调控基因转录的重要转录因子，这将有望为肝癌的诊断和治疗提供新的治疗靶点。
　　목적감정정합소β6기인재HepG2세포중적주요전록조공구，위진일보연구αvβ6재간암발생발전과정중적전록조공궤제전정기출。방법이용β6기인전록기시위점부근1135 bp기인서렬경PCR진행불동장도적결실확증，병련접도pGL2-basic재체，구건중조질립，이용쌍형광소매보고기인분석계통검측계동자적전록활력，병이용Transcription Element Search System예측β6기인주요전록조공구잠재전록인자결합위점。결과획득4개함불동장도β6기인서렬적중조질립；당β6기인재거전록기시위점-458 bp절단도-187 bp시，전록활력강저약50％，재저개전록조공구존재YY1、AP-1화c-Myb등여간암상관적전록인자결합위점。결론재HepG2세포중，β6기인계동자구적-458 bp/-187 bp구역시기주요적전록조공구，YY1、AP-1화c-Myb삼개전록인자존재해구역，병가능시조공기인전록적중요전록인자，저장유망위간암적진단화치료제공신적치료파점。
Objective To identify key transcriptional regulatory regions of human integrin beta6 (Itgβ6) gene in HepG2 cells, thus to make a preliminary study for its transcriptional regulatory mechanism. Methods The recombinant plasmids containing different lengths of DNA fragments upstream of translation initiation site of Itgβ6 gene as reporter promoters were constructed by deletion Method, and the promoter activities were detected using dual luciferase reporter assay system. The potential transcription factor binding sites at key transcriptional regulatory region of Itgβ6 gene were predicted by TESS. Results The recombinant plasmids containing different lengths of Itgβ6 gene were obtained. When the lengths of Itgβ6 gene were reduced from-458 bp to-187 bp, the transcriptional activity was decreased by about 50%. There were YY1、AP-1 and c-Myb transcriptional factor binding sites in the-458 bp to-187 bp regions. Conclusion The-458 bp/-187 bp was the key transcriptional regulatory region of Itgβ6 gene in HepG2 cells, YY1、AP-1 and c-Myb are in this region, and may be the important factors for regulating the transcription of Itgβ6 gene.