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已有的报告基因和EMSA实验研究表明 ,人干细胞生长因子 (SCF)基因 5′旁侧AT富集区- 1 1 90～ - 853在HeLa和MCF 7细胞中均能增强下游基因转录 ,可能为一个核基质结合区(MAR) ,对人SCF基因的转录发挥调控作用 .为进一步研究该AT富集区的功能 ,将人SCF基因 5′旁侧 - 1 1 90～ - 853AT富集区分别克隆入SV40或CMV启动子前后紧接着CAT报告基因 ,瞬时转染Jurkat,HepG2和 3T3细胞 ,检测CAT报告基因的瞬时表达活性 .结果表明 :人SCF基因 5′旁侧- 1 1 90～ - 853AT富集区在Jurkat和HepG2细胞中 ,对分别由SV40和CMV启动子引导的CAT基因表达均有抑制作用 ;但在 3T3细胞中对SV40启动子的转录活性表现出增强作用 ,对CMV启动子的转录活性无明显影响 .这些结果提示 ,人SCF基因 5′旁侧 - 1 1 90～ - 853AT富集区转录调控具有组织细胞特异性 ,在不同的细胞中可能发挥转录增强或抑制作用
The existing reporter gene and EMSA experimental studies have shown that AT enrichment region - 1 1 90 ~ - 853 next to the 5 ’side of the human stem cell growth factor (SCF) gene can enhance downstream gene transcription in HeLa and MCF 7 cells, which may be A nuclear matrix-binding domain (MAR), regulates the transcription of human SCF gene.To further investigate the function of this AT-rich region, the 5 ’flanking-flanking 1 1 90-853AT enrichment region of human SCF gene was separately cloned CAT reporter gene was cloned immediately before and after SV40 or CMV promoter and transiently transfected into Jurkat, HepG2 and 3T3 cells to detect the transient expression activity of CAT reporter gene.The results showed that the 5 ’flank of human SCF gene - 1 1 90 ~ - 853AT rich The Jurkat and HepG2 cells inhibited the CAT gene expression mediated by both the SV40 and CMV promoters, but enhanced the transcriptional activity of the SV40 promoter in 3T3 cells, and the transcription of the CMV promoter These results suggest that the transcriptional regulation of 5 ’flanking - 1 1 90 ~ - 853AT enrichment region of human SCF gene is tissue-cell specific and may play an important role in the transcription enhancement or inhibition in different cells