BAY11-7082通过抑制三磷酸腺苷柠檬酸裂解酶磷酸化抑制乳腺癌细胞增殖并促进其凋亡  

BAY11-7082 inhibits proliferation and promotes apoptosis in breast carcinoma MCF-7 cells by inhibiting phosphorylation of ATP citrate lyase

作　　者：黄云秀[1] 苏敏[1] 魏兰[1] 季飞虎[1] 王念[1] 钟昌莉 陈婷梅[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《细胞与分子免疫学杂志》2015年第11期1458-1462,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81272544);重庆市自然科学基金(cstc2012jj A10011)

摘　　要：目的探讨核因子κB信号通路抑制剂BAY11-7082通过调控三磷酸腺苷柠檬酸裂解酶(ACL)对MCF-7乳腺癌细胞增殖及凋亡的影响。方法选取对数生长期的MCF-7细胞,随机分为对照组和5μmol/L BAY11-7082处理组、10μmol/L LY294002处理组。采用Western blot法检测BAY11-7082和LY294002处理后MCF-7细胞中ACL、磷酸化的ACL(p-ACL)、磷酸化的Akt(p-Akt)、磷酸化的核因子κB(p-NF-κB)的蛋白水平。采用BAY11-7082和小干扰RNA敲低ACL(si ACL),CCK-8法检测MCF-7细胞的增殖、异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测MCF-7细胞的凋亡。结果 BAY11-7082能抑制MCF-7细胞增殖,且呈剂量依赖性。Werstern blot结果显示BAY11-7082处理MCF-7细胞48 h后,p-NF-κB和p-ACL明显下降;LY294002处理组中,p-Akt和p-ACL明显下降。CCK-8实验和流式细胞术检测结果显示,BAY11-7082和si ACL分别处理MCF-7细胞48 h后,细胞增殖减少,细胞凋亡明显增加。结论 BAY11-7082能通过抑制ACL的磷酸化从而抑制乳腺癌MCF-7细胞的增殖并促进其凋亡。Objective To investigate the effect of NF-KB inhibitor BAY11-7082 on proliferation and apoptosis of breast carcinoma MCF-7 cells and the underlying mechanism. Methods MCF-7 cells in the logarithmic growth phase were divided into control group, 5 μmol/L BAY11-7082 group and 10 μmol/L LY294002 group. After the treatment of BAY11-7082 and LY294002, the protein levels of ATP citrate lyase （ACL）, phosphated-ACL （p-ACL）, phosphated-Akt （p-Akt） and phosphatecl nuclear factor KB （p-NF-κB） were determined by Western blotting. The proliferation of MCF-7 cells treated with BAY11-7082 or siACL were detected by CCK-8 assay. The apoptosis of MCF-7 cells treated with BAYⅡ-7082 or siACL were observed by flow cytometry combined with annexin V-FITC/PI staining. Results The proliferation of MCF-7 cells was inhibited by BAY11-7082 in a dose-dependent manner. Compared with the control group, the expressions of p-ACL and p-NF-ⅡB protein in MCF-7 cells treated with BAY11-7082 were lowered. The expressions of p-ACL and p-NF-KB protein in MCF-7 cells treated with lO pmol/L LY294002 were also reduced significantly. The proliferation of MCF-7 cells treated with BAY11-7082 or siACL for 48 hours was inhibited and the apoptosis was promoted significantly, as shown by CCK-8 assay and flow cytometry. Conclusion BAY11-7082 could inhibit the proliferation of MCF-7 breast carcinoma cells and promote the apoptosis by inhibiting the phosphorylation of ACL.

关 键 词：BAY11-7082 LY294002 ATP柠檬酸裂解酶 MCF-7细胞 增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤] R392-33[医药卫生—临床医学]