孕激素对人子宫内膜样癌JEC细胞ER亚型、PR亚型及p57^kip2表达的影响  

Effects of progesterone on the expressions of ER subtype,PR subtype and P57^(kip2) in JEC cells of human endometrial carcinoma

作　　者：刘蒙蒙[1,2] 孙小杰 周正平[3] 袁丹 刘俊江[1] Liu Mengmeng1,2, Sun Xiaofie1, Zhou Zhengping3, Yuan Dan1 , Liu Junfiang1(1. Department of Pathology,Zunyi Medical University, Zunyi Guizhou 563099, China ; 2. Department of Patholo- gy of Binzhou People＇ s Hospital, Binzhou Shandong 256610, China; 3. Electron Microscope Laboratory of Basic Medical College, Zunyi Medical University,Zunyi Guizhou 563099, Chin)

机构地区：[1]遵义医学院病理学教研室,贵州遵义563099 [2]滨州市人民医院病理科,山东滨州256610 [3]遵义医学院基础医学院电镜室,贵州遵义563099

出　　处：《遵义医学院学报》2018年第1期46-52,共7页Journal of Zunyi Medical University

基　　金：贵州省社发攻关项目(黔科合SY字[2013]3011)

摘　　要：目的研究孕激素干预治疗对人子宫内膜样癌JEC细胞中雌激素受体(estrogen receptor,ER)两种亚型、孕激素受体(progesterone receptor,PR)两种亚型及p57^(kip2)蛋白表达的影响。方法实验组:分别用3种不同浓度的孕酮(Progesterone,P)(10^(-4)mol/L、10^(-6)mol/L、10^(-8)mol/L)干预体外培养的中分化JEC细胞;对照组:用不含药物的等量培养基培养JEC细胞。分别培养24、48、72、96h后,用MTT法观察JEC细胞的生长情况;培养24、48、72h后,在光镜、电镜下观察JEC细胞的生长情况及超微结构变化,用免疫蛋白印迹(Western Blot,WB)法检测JEC细胞中ER亚型(ERα、ERβ)、PR亚型(PRA、PR-B)及p57^(kip2)蛋白的表达情况。结果光镜及MTT结果显示,P低、中浓度组可促进JEC细胞的生长,高浓度组则抑制JEC细胞的生长(P<0.05)。电镜结果显示,P各浓度组胞质内均可见自噬现象,高浓度组细胞表面微绒毛及胞质内分泌泡均增多。WB结果显示,随着药物作用浓度的增高,p57^(kip2)蛋白表达逐渐增加,ERβ蛋白在24、72h表达逐渐增加(P<0.05);随着药物作用时间的延长,p57^(kip2)蛋白在各组表达均逐渐增高,ERβ蛋白在高浓度组表达逐渐增高,在低、中浓度组则以72h表达最高,48h表达最低(P<0.05);对照组及P各组JEC细胞均无ERα、PR-A、PR-B蛋白的表达。结论孕酮可上调JEC细胞中ERβ、p57^(kip2)蛋白的表达,低浓度孕酮以ERβ蛋白促细胞生长为主,高浓度孕酮以p57^(kip2)蛋白阻滞细胞周期进程为主导。JEC细胞不表达ERα、PR-A、PR-B蛋白,且P干扰刺激后不能诱导其表达。联合检测ERα、ERβ、PR-A、PR-B、p57^(kip2)蛋白在EC中的表达,有利于评估EC的发生发展。Objective To explore the effects of progesterone on the expression levels of two types of estrogen receptor subtypes,two types of progesterone receptor subtypes and p57^（kip2） protein in human endometrioid carcinoma cell JEC. Methods The moderately differentiated JEC cells were treated with different concentrations of progesterone（ P）（ 10-4,10-6 and 10-8 mol/L） in vitro respectively. The control group cells were cultured in the culture medium without drug. MTT was used to detect the growth of JEC cells after 24,48,72,96 h. The growth situation and the morphological changes of JEC cells were observed by the inverted microscope and transmission electron microscope. Western Blot（ WB） was used to detect the expression levels of ER subtypes（ ERα,ERβ）,PR subtypes（ PR-A,PR-B） and p57^（kip2） protein in JEC cells after 24,48,72 h. Results The low and medium concentrations of P could promote the growth of JEC cells,while the high concentration of P could inhibit the growth of JEC cells（ P〈0. 05）. From the results of electron microscopy,autophagy was found in the cytoplasm of P groups,microvilli and endocrine vesicles in the high concentration group were increased. With the increase of the concentration of P,the level of p57^（kip2） protein was increased,and the ERβ protein level was increased gradually at 24,72 h（ P〈0. 05）. With the extension of the interference time,the p57^（kip2） protein level was increased,the level of ERβ protein was increased in the high concentration group,and the level of ERβ protein was the highest at72 h and the lowest at 48 h in the low and medium concentration groups（ P〈0. 05）. ERα,PR-A,and PR-B protein were not detected in the control group and P groups. Conclusion P can increase the levels of ERβ and p57^（kip2） protein in JEC cells. In the low and medium concentration groups,ERβ can promote the JEC cell growth,while in the high concentration group,p57^（kip2） can block cell cycle progression,and inhibit the growth of JEC cells

关 键 词：子宫内膜样癌 孕激素 ER PR P57^KIP2 

分 类 号：R737.33[医药卫生—肿瘤]