中国医科大学学报2017，Vol.46Issue(8)：677-680,685,5.DOI:10.12007/j.issn.0258-4646.2017.08.002
乳腺癌MCF-7细胞p21WAF1/CIP1启动子区雌激素受体α的高功能结合位点
High-affinity Binding Sites for Estrogen Receptor α in the p21WAF1/CIP1 Promoter Region in Breast Cancer MCF-7 Cells
摘要
Abstract
Objective To investigate the specific sites that estrogen receptor (ER)α could be recruited to in the p21WAF1/CIP1 promoter region to regulate its transcriptional activity in MCF-7 cells,and to clarify the molecular mechanism of suberoylanilide hydroxamic acid (SAHA) and leptin in the regulation of p21WAFI/CIP1 promoter function.Methods MCF-7 cells were starved by culturing them in fetal calf serum-free medium for 24 hours,and then treated with 20 μmol/L of 0.88 μL SAHA (SAHA group) or 0.625 nmol/L of 10 μL leptin (leptin group) for 24 hours,or cultured in complete RPMI-1640 medium (control group).Cell lysates were incubated with anti-ERα antibody for ChIP analysis.The relative expression levels of DNA fragments,ranging from the TSS to upstream of the p21WAF1/CIP1 promoter region (+2 to-4 000 bp),that bound the antibody were detected by real-time PCR.Results In the control group,the relative expression levels of f1,f2,and f8 DNA fragments that bound the antiERα antibody were two-fold higher than the relative expression of the f9 fragment (P ＜ 0.01).In the SAHA and leptin groups,the relative expression of f1 to f10 DNA fragment that bound anti-ERα antibody was significantly lower than that of the control.The binding affinity of ERα for the f8 fragment was the lowest (P ＜ 0.01) in the SAHA group,and it was significantly lower than that in the leptin group (P ＜ 0.01).Conclusion ERα could be recruited to the p21WAFI/CIP1 promoter via signaling pathways activated during the proliferation of breast cancer MCF-7 cells.Moreover,the DNA fragment ranging from-2 800 to-3 200 bp upstream of the p21 WAF1/CIP1 promoter is the target functional region for high-affinity binding with ERα.关键词
乳腺癌/MCF-7细胞/p21WAF1/CIP1/雌激素受体α/辛二酰苯胺异羟肟酸/瘦素Key words
breast cancer/MCF-7 cell/p21WAF1/CIP1/estrogen receptor α/suberoylanilide hydroxamic acid/leptin分类
医药卫生引用本文复制引用
邹丹,冯秀艳,周伟强..乳腺癌MCF-7细胞p21WAF1/CIP1启动子区雌激素受体α的高功能结合位点[J].中国医科大学学报,2017,46(8):677-680,685,5.基金项目
国家自然科学基金(81172509) （81172509）
辽宁省自然科学基金(201602735) （201602735）
沈阳市科学技术计划(F15-199-1-28) （F15-199-1-28）
