PRB介导子宫内膜癌细胞对MPA敏感性的作用机制研究  

Mechanism study of PRB in regulating therapeutic sensitivity of MPA in endometrial cancer cells

作　　者：王晶[1] 孙笑[1] 王丽华[1] 王玉东[1] 

机构地区：[1]上海交通大学医学院附属国际和平妇幼保健院妇科,上海200030

出　　处：《中国药理学通报》2014年第9期1252-1256,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81172477);上海市科委自然科学基金项目(No 11ZR1440800;13JC1401303);上海市卫生系统优秀学科带头人培养计划资助项目(NoXBR2013097)

摘　　要：目的明确孕激素受体B(PRB)介导子宫内膜癌细胞对醋酸甲羟孕酮(MPA)的敏感性,及观察干扰PRB表达,MPA对内膜癌细胞生物学行为的影响。方法利用慢病毒载体系统转染对孕激素敏感的Ishikawa细胞,干扰PRB表达,实时荧光定量PCR(qPCR)及Western blot检测Ishikawa细胞PRB的mRNA和蛋白水平。MTT法、流式细胞技术(FACS)、Transwell法分别检测MPA对内膜癌细胞增殖、凋亡及侵袭能力的影响;Western blot检测MPA对子宫内膜癌细胞丝裂原活化蛋白激酶(MAPK)/细胞外信号调节激酶(ERK)信号通路的影响。结果慢病毒载体介导的RNA干扰Ishikawa细胞明显抑制了PRB的mRNA和蛋白表达水平(P<0.01);MPA对Ishikawa细胞具有抑制增殖、侵袭,并促进凋亡的作用(P<0.01);干扰PRB表达后,孕激素作用则相反,且激活MAPK/ERK通路。结论 PRB介导了子宫内膜癌细胞对MPA的敏感性,干扰PRB的表达,MPA可通过激活ERK1/2-MAPK通路,影响内膜癌细胞的生物学行为。Aim To explore the role of progesterone re-ceptor B （ PRB ） in regulation of medroxyprogesterone acetate （ MPA） sensitivity in endometrial cancer cells, and to investigate the effect of MPA on the biological character in Ishikawa cells infected with shRNA targe-ting PRB gene. Methods Ishikawa cells were stably transfected with PRB shRNA using lentivirus to knock-down endogenous PRB expression. Real-time fluores-cent quantitative PCR was applied to confirm the knockdown effect. MTT assay, flow cytometry and cell invasion assay were applied to detect the influence of MPA on endometrial cancer cell proliferation, apopto-sis and invasion. We also used Western blot assay to detect the effect of MPA induced the activation of ERK/MAPK signal pathway. Results Recombinant lentiviral vector expressing shRNA targeting PRB gene was successfully established, results of real-time PCR and Western blot showed that compared with control group, PRB expression in Ishikawa cells infected with shRNA decreased obviously （ P 〈0.01 ）; MPA could repress endometrial cancer cells proliferation and inva-sion, meanwhile promoted its apoptosis （ P 〈0.01 ） . However, the effect was almost reverse in Ishikawa cells infected with shRNA. Furthermore, MPA induced ERK/MAPK activation in Ishikawa cells infected with shRNA. Conclusions PRB plays a role in regulating therapeutic sensitivity of MPA in endometrial cancer cells;and for Ishikawa cells infected with shRNA tar-geting PRB gene, MPA has effects on the biological character via pERK1/2-MAPK signal pathway.

关 键 词：子宫内膜癌细胞 丝裂原活化蛋白激酶 细胞外信号调节激酶 醋酸甲羟孕酮 孕激素受体B 孕激素耐药 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R345.57[医药卫生—基础医学]