中国药理学通报2017，Vol.33Issue(3)：317-321,5.DOI:10.3969/j.issn.1001-1978.2017.03.006
乳腺癌MCF-7细胞p21 WAF1/CIP1启动子区HDAC1高功能结合位点的研究
A study on high function binding site of HDAC1 in p21 WAF1/CIP1 promoter region in breast cancer MCF-7 cells
摘要
Abstract
Aim Toinvestigatethespecificbinding sites that HDAC1 can be recruited to regulate the tran-scriptional activity of p21 WAF1/CIP1 promoter in the breast cancer MCF-7 cells.Methods ThebreastcancerMCF-7 cells in logarithmic growth phase were starved with FBS free medium for 24 hours,and treated with 20 μmol·L-1 SAHA(S group)or 0. 625 nmol·L-1 Leptin(L group)for 24 hours,and the cells that were cultured in the complete RPMI 1640 medium without any treatment were assigned as control group (B group).The DNA-ChIP was followed the manufactur-er′s protocol for the assay.The cell lysis was prepared and incubated with anti-HDAC1 antibody overnight at 4℃.DNA fragments binding anti-HDAC1 antibody were gathered and purified.The relative expression level of DNA fragments from TSS to the upstream of the p21 WAF1/CIP1 promoter region(+2 ~-4000 bp)bind-ing with antibody was detected by Real-time PCR and analyzedby2-ΔΔCTmethod.Results InBgroup, HDAC1 had high affinity with the f1 and f 8 fragmentsof p21 WAF1/CIP1 promoter compared to the other fragemts,and showed the highest affinity with the f8 fragment.In S group,the binding ability of HDAC1 to the f1 ~f10 fragment of p21 WAF1/CIP1 promoter was sig-nificantly lower than that of the control.The binding activity of HDAC1 to f8 fragment was the lowest,while reversing to reach the peak after leptin treatment.Con-clusions HDAC1canberecruitedtop21WAF1/CIP1pro-moter by the cell proliferation signal during the prolifer-ation of breast cancer MCF-7 cells.The DNA fragment from -2800 to -3200 bp in the upstream of p21 WAF1/CIP1 promoter is the target functional region for the binding with HDAC1 .关键词
乳腺癌/MCF-7细胞/p21WAF1/CIP1/组蛋白去乙酰化酶1/辛二酰苯胺异羟肟酸/瘦素Key words
breast cancer/MCF-7 cell/p21 WAF1/CIP1/HDAC1/suberoylanilide hydroxamic acid/leptin分类
医药卫生引用本文复制引用
邹丹,周伟强..乳腺癌MCF-7细胞p21 WAF1/CIP1启动子区HDAC1高功能结合位点的研究[J].中国药理学通报,2017,33(3):317-321,5.基金项目
国家自然科学基金资助项目(No 81172509) （No 81172509）
辽宁省自然科学基金资助项目(No 201602735) （No 201602735）
沈阳市科技计划项目(No F15-199-1-28) （No F15-199-1-28）
