摘要
目的探讨赫赛汀联合9-顺式视黄酸对ErbB2阳性乳腺癌细胞周期进程及分布的影响及其相应的分子机制。方法利用流式细胞仪检测经赫赛汀、9-顺式视黄酸单独和联合作用后,癌基因(ErbB2/neu)阳性的MDA-MB-453乳腺癌细胞周期进程和分布的变化。在此基础上,利用免疫印记和半定量PCR法对CDK2、CyclinE、P27的表达进行检测,同时利用免疫沉淀法检测CyclinE/CDK2结合力的变化。结果与对照组比较,一定剂量的赫赛汀和9-顺式视黄酸可将MDA-MB-453细胞的周期进程阻遏于G0/G1期从而抑制其增殖活性。经2者联合作用后,CyclinE、CDK2的表达较对照组均有所下降,其中以CDK2的变化最为明显,而P27蛋白表达则明显升高。同时,2者联合作用还能有效降低CyclinE/CDK2的结合能力。结论赫赛汀和9-顺式视黄酸可在体外分别通过改变CyclinE、CDK2和P27的表达和活性起到协同抑制HrbB2/neu阳性乳腺癌的目的。
Objective To explore the synergistic effects of herceptin and 9-cis RA on the cell cycle progression of HER2/neu-positive breast cancer cells and associated molecular mechanism.Methods After the treatment of MDA-MB-453 cells with herceptin and 9-cis RA,flow cytometry assay was employed to detect the inhibitory effects on cell cycle progression.After that,mRNA and protein expression of cyclin E,CDK2,and P27 were analyzed by RT-PCR and Western blotting.At the same time,the bingding affinity of cyclin E and CDK2 was tested by immunoprecepitation assay.Results Herceptin and 9-cis RA synergistically blocked the cell cycle progression of MDA-MB-453 cells in G0/G1 phase in vitro.The expression of cyclin E and CDK2 were notably down-regulated as well as the cyclin E/CDK2 binding affinity,while the P27 protein expression was remarkably up-regulated by the co-treatment.Conclusion Herceptin and 9-cis RA could exert synergistic inhibitory effects on cell cycle progression of ErbB2-positive breast cancer cells by influencing the expression and activity of cyclin E,CDK2 and P27.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2008年第3期311-312,共2页
Chinese Journal of Public Health
基金
国家自然科学基金(30300285)
