雌激素对人子宫内膜样癌Ishikawa细胞中p57^(kip2)、Cyclin D1、CDK4表达及形态变化影响的研究  

Effects of estrogen on the expressions of p57^(kip2),Cyclin D1,CDK4 and morphological changes in Ishikawa cells in human endometrial carcinoma

作　　者：袁丹[1] 刘蒙蒙[1] 周正平[2] 刘俊江 孙小杰[1] Yuan Dan;Liu Mengmeng;Zhou Zhengping(Department of Pathology;Department of Electron Microscopy, Zunyi Medical College ,Zunyi 563099)

机构地区：[1]遵义医学院病理学教研室,遵义563099 [2]遵义医学院电镜室,遵义563099

出　　处：《现代妇产科进展》2018年第4期249-254,共6页Progress in Obstetrics and Gynecology

基　　金：贵州省社发攻关项目(黔科合SY字[2013]3011);贵州省科技计划项目(黔科合基础[2017]1216)

摘　　要：目的:研究雌激素对人子宫内膜样癌(EC)Ishikawa细胞(高分化)中p57^(kip2)、Cyclin D1、CDK4表达及形态变化的影响。方法:分别采用含3种不同浓度雌二醇(E_2)(10^(-6)、10^(-8)、10^(-10)mol/L)的培养基培养Ishikawa细胞24、48、72h,MTT检测细胞增殖情况,光镜和电镜观察细胞形态变化,Western blot法检测p57^(kip2)、Cyclin D1、CDK4蛋白表达。结果:MTT结果显示,与对照组比较,低、中浓度E_2组Ishikawa细胞增殖明显(P<0.05),高浓度E_2组细胞增殖不明显(P>0.05)。光镜及电镜结果显示,与对照组比较,低、中浓度E2组Ishikawa细胞密度增加,部分细胞内线粒体、内质网轻微肿胀,随着作用时间延长,其肿胀程度增加;高浓度E_2组Ishikawa细胞密度减少,较多细胞内线粒体明显肿胀,可见细胞凋亡现象,随着作用时间延长,线粒体、内质网弥漫肿胀、空泡化,细胞凋亡增多。Western blot法结果显示,随着E_2作用时间延长及浓度增加,p57^(kip2)、Cyclin D1与CDK4蛋白表达均逐渐增加(P<0.05),其中p57^(kip2)、Cyclin D1、CDK4蛋白均在高浓度E_2组表达最高,而p57^(kip2)蛋白在低浓度E_2组表达最低,Cyclin D1与CDK4蛋白在对照组表达最低。Cyclin D1和CDK4蛋白表达呈正相关(P<0.05)。结论:E_2浓度较低时可能以上调Cyclin D1与CDK4蛋白表达为主,发挥正向调控细胞周期进程的作用,促进Ishikawa细胞增殖;E_2浓度较高时可能以上调p57^(kip2)蛋白表达为主,从而抑制Ishikawa细胞增殖,还可导致较强细胞损伤。Objective： To study the effects of estrogen on the expressions of p57^（kip2）,Cyclin D1 and CDK4 and morphological changes in endometrial carcinoma（ EC） cells named Ishikawa（ high differentiation） in vitro.Methods： The Ishikawa cells cultured in vitro were treated with three different concentrations（ 10-6 mol/L,10-8 mol/L,10-10 mol/L） of estradiol（ E_2）,after 24,48,72 hours.The cells proliferation was measured by MTT.Light and electron microscopic was observed on morphological changes of Ishikawa cells. Western blot was used to detect the expressions of p57^（kip2）,Cyclin D1 and CDK4 protein in Ishikawa cells. Results： The results of MTT showed that compared with the control group,the medium and low concentration of E_2 group promoted the proliferation of Ishikawa cells obviously（ P0.05）.The proliferation of cells in high concentration of E_2 group was not obvious（ P 0.05）. Light and electron microscopic：compared with the control group,the density of low concentration of E_2 group increased in Ishikawa cells.Some cells of mitochondria and endoplasmic reticulum was swell slightly. With theextension of the time,the swelling degree increased. The density of high concentration of E_2 group was reduced in Ishikawa cells,swelling of mitochondria in many cells obviously,and visible cell apoptosis. With the prolongation of time,mitochondria,endoplasmic reticulum was diffuse swelling and vacuolization,the cell apoptosis increased. Western blot results showed that with the increase in the concentration of E_2 and the extension of the time,the expressions of p57^（kip2）,Cyclin D1 and CDK4 protein were increased（ P0.05）.Including the expressions of p57^（kip2）,Cyclin D1,CDK4 protein were the highest in high concentration of E_2 group.p57^（kip2） protein was the lowest in the low concentration of E_2 group. Cyclin D1 and CDK4 protein was the lowest in the control group.There was a positive correlation between the expression of Cyclin D1 and CDK4 protein（ P 0.05）. Conclusi

关 键 词：子宫内膜癌 ISHIKAWA细胞 雌激素 P57KIP2 Cyclin D1 CDK4 

分 类 号：R737.33[医药卫生—肿瘤]