ER拮抗剂对Ishikawa细胞中p57^(kip2)、Cyclin D1表达及形态变化的影响研究  

Effects of ER antagonists on the expression of p57^(kip2),Cyclin D1 and morphological changes in Ishikawa cells

作　　者：袁丹[1,2] 刘俊江 周正平 刘蒙蒙[1] Yuan Dan;Liu Junjiang;Zhou Zhengping(Department of Pathology,Zunyi Medical College,Zunyi 563099;Department of Electron Microscopy,Zunyi Medical College,Zunyi 563099;Department of Pathology,Affiliated Hospital 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]遵义医学院附属医院病理科,遵义563099 [3]遵义医学院电镜室,遵义563099

出　　处：《现代妇产科进展》2019年第1期6-12,共7页Progress in Obstetrics and Gynecology

基　　金：贵州省科技计划项目(黔科合基础[2017]1216)

摘　　要：目的:研究雌激素受体(ER)拮抗剂对人子宫内膜样癌(EC) Ishikawa细胞(高分化)形态变化及细胞中p57^(kip2)、Cyclin D1、CDK4表达情况的影响,探讨女性激素调控、细胞周期调控和EC发生、发展的影响。方法:分别用含雌二醇(E2)、他莫昔芬(TAM)、氟维司群(Faslodex,ICI182780)、E2+TAM、E2+ICI182780的培养基培养Ishikawa细胞,用不含药物的培养基作为对照。培养24、48、72、96h后,MTT法观察细胞增殖情况,培养24、48、72h后,通过光镜和电镜观察细胞的形态变化,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,E2组、ICI182780组、E2+ICI182780组Ishikawa细胞增殖活性降低,以E2+ICI182780组更甚(P<0.05);与E2组比较,E2+ICI182780组的细胞增殖活性降低(P<0.05);实验组及对照组细胞的增殖活性均随着药物作用时间延长而增强(P<0.05)。光镜下:与对照组比较,E2组、ICI182780组、E2+ICI182780组细胞密度降低,以E2+ICI182780组更甚,TAM组则稍增加;与E2组比较,E2+ICI182780组细胞密度降低,E2+TAM组则增加;各组细胞形态变化均不明显。电镜下:与对照组比较,E2组部分细胞可见内质网、线粒体明显肿胀,其余各组内质网呈不同程度肿胀,可见凋亡现象;与E2组比较,TAM组、ICI182780组内质网肿胀较轻,E2+TAM、E2+ICI182780组肿胀明显。Western blot结果示:与对照组比较,随着药物作用时间延长,p57^(kip2)蛋白在E2组、ICI182780组、E2+ICI182780组表达逐渐增加,其中E2+ICI182780组表达最高(P <0. 05); Cyclin D1、CDK4蛋白则在E2组、ICI182780组、E2+ICI182780组表达逐渐降低,其中E2+ICI182780组表达最低(P <0. 05);与E2组比较,p57^(kip2)蛋白在E2+ICI182780组表达增加,在E2+TAM组表达降低(P<0.05); Cyclin D1、CDK4蛋白在E2+ICI182780组表达降低,在E2+TAM组表达增加(P<0.05)。结论:ER拮抗剂ICI182780单独或联合使用雌激素,可能通过诱导p57^(kip2)蛋白表达,下调Cyclin D1、CDK4蛋Objective:To study the effects of estrogen receptor antagonist on the expressions of p57^kip2,Cyclin D1 and CDK4 protein and the morphological changes in human endometrioid carcinoma cells(EC)named Ishikawa(highly differentiated)in vitro,which to explore hormonal regulation,cell cycle regulation on the occurrence and development of EC.Methods:The Ishikawa cells cultured in vitro were treated with estradiol(E2),Tamoxifen(TAM),fulvestrant(ICI182780),E2+TAM,E2+ICI182780 and another group was cultured without any drugs as control group.After 24,48,72,96h,the cells proliferation and morphological changes were measured by MTT.After 24,48,72h,the cells morphological changes were measured by light and electron microscopic.Western blot was used to detect the expressions of p57^kip2,Cyclin D1 and CDK4 protein in cells.Results:The results of MTT showed that compared with the control group,the E2 group,ICI182780 group and E2+ICI182780 group restrained the proliferation of Ishikawa cells,and E2+ICI182780 group was more obvious(P<0.05).Compared with the E2 group,E2+ICI182780 group restrained the proliferation of Ishikawa cells(P<0.05).The proliferative activity of the experimental groups and the control group increased with the prolongation of the time of action of the drug(P<0.05).Light microscope:Compared with the control group,the density of E2 group,ICI182780 group and E2+ICI182780 group decreased in Ishikawa cell,the E2+ICI182780 group was more obvious.The density of the TAM group increased slightly.compared with E2 control group,the density of E2+ICI182780 group decreased in Ishikawa cell.The density of E2+TAM group increased.The morphological changes of cells in each group were not obvious.Electron microscope:compared with the control group,a part of E2 group cells of endoplasmic reticulum and mitochondria were swollen.The other groups dominated by vary degrees of endoplasmic reticulum swelling,and seen apoptotic phenomenon.Compared with E2 group,the TAM group and ICI182780 group endoplasmic reticulum swelling and light

关 键 词：子宫内膜癌 ER拮抗剂 P57^KIP2 CYCLIND1 CDK4 

分 类 号：R737.33[医药卫生—肿瘤]