雌激素对人子宫内膜样癌Ishikawa细胞增殖、形态结构的影响及可能的作用机制  

Effects of estrogen on proliferation and morphological structure of human endometrioid carcinoma Ishikawa cells,and the possible mechanism

作　　者：刘俊江 周正平 袁丹 刘蒙蒙 Liu Junjiang;Zhou Zhengping;Yuan Dan;Liu Mengmeng(Department of Pathology,Zunyi Medical University,Zunyi Guizhou 563099,China;Electron Microscopy Laboratory,Zunyi Medical University,Zunyi Guizhou 563099,China)

机构地区：[1]遵义医科大学病理学教研室,贵州遵义563099 [2]遵义医科大学电镜室,贵州遵义563099

出　　处：《遵义医学院学报》2019年第1期45-51,共7页Journal of Zunyi Medical University

基　　金：贵州省科技计划资助项目(NO:黔科合基础[2017]1216)

摘　　要：目的探讨雌激素(estrogen,E)对人高分化子宫内膜样癌Ishikawa细胞增殖、形态结构的影响及可能的作用机制。方法予浓度为10^(-4)、10^(-2)、1μmol/L的雌二醇(β-Estradiol,E_2)处理(设不含药物的等体积完全培养基为对照组)体外培养的Ishikawa细胞,采用MTT检测细胞生长情况,光镜、电镜观察细胞形态结构改变,Real-time PCR法(RT-PCR)、Western blot法(WB)分别检测细胞中p57^(kip2)和Cyclin E mRNA及蛋白表达水平。结果 MTT:与对照组相比,1μmol/L组Ishikawa细胞增殖能力减弱,余实验组细胞增殖能力则增强(P<0.05)。形态改变:与对照组相比,10^(-4)、10^(-2)μmol/L组细胞密度增加(以10^(-2)μmol/L组更明显),在各实验组细胞内可见线粒体、内质网肿胀(以1μmol/L组更明显)。RT-PCR及WB:随作用时间延长,p57^(kip2)mRNA在1μmol/L组表达逐渐增加,在10^(-4)、10^(-2)μmol/L组表达则逐渐降低,其中,在1μmol/L组表达最高、在10^(-2)μmol/L组表达最低,p57^(kip2)蛋白在各实验组表达则随E_2作用浓度增加而增加(P<0.05);Cyclin E mRNA及蛋白在1μmol/L组表达降低,在10^(-4)、10^(-2)μmol/L组表达则增加,其中,在1μmol/L组表达最低、在10^(-2)μmol/L组表达最高(P<0.05)。结论在Ishikawa细胞中,较低浓度E可能通过减少p57^(kip2)表达、增加Cyclin E表达,促使细胞通过G_1期,从而增强了细胞增殖能力;而较高浓度E则可能通过诱导p57^(kip2)表达、减少Cyclin E表达,阻碍细胞通过G_1期,还使细胞损伤明显,达到阻碍肿瘤进展的目的。Objective To investigate the effects of estrogen (E) on the proliferation and morphological structure of highly differentiated human endometrioid carcinoma Ishikawa cells,and try to find the possible mechanism. Methods Ishikawa cells were cultured in vitro and treated with β-Estradiol (E 2) at the concentration of 10 ^-4 ,10 ^-2 ,1 μmol/L,respectively.Control group was cultured in an equal volume of complete medium without drug.MTT was used to detecte the cell growth.Light and electron microscopes were used to observe the changes of cell morphology.Western blot and Real-time PCR were used to detecte the expression of p57 kip2 and Cyclin E mRNA and protein. Results MTT:Compared with the control group,the proliferation of Ishikawa cells in the 1 μmol/L group was weakened,and the proliferation ability of the remaining experimental groups was enhanced ( P <0.05).Morphological changes:Compared with the control group,the cell density increased in the 10^-4 and 10 ^-2 μmol/L groups (it was more obvious in the 10 ^-2 μmol/L group),and the mitochondria and endoplasmic were seen to be swollen in the cells of each experimental group (it was more obvious in the 1 μmol/L group).RT-PCR and WB:With the prolongation of the action time,the expression of p57 kip2 mRNA gradually increased in the 1 μmol/L group,but gradually decreased in the 10 ^-4 and 10 ^-2 μmol/L groups.The expression was highest in the 1 μmol/L group and lowest in the 10 ^-2 μmol/L group.While,the expression of p57 kip2 protein in each experimental group increased with the increase of E 2 concentration ( P <0.05).The expression of Cyclin E mRNA and protein decreased in the 1 μmol/L group,but increased in the 10 ^-4 and 10^-2 μmol/L groups,with the lowest expression in the 1 μmol/L group and the highest in the 10 ^-2 μmol/L group ( P <0.05). Conclusion In Ishikawa cells,lower concentrations of E may increase the expression of Cyclin E and decrease the expression of p57 kip2 ,promote cells passage through G 1 phase rapidly,and enhance cell proliferat

关 键 词：子宫内膜样癌 ISHIKAWA细胞 雌二醇 P57KIP2 CYCLINE 

分 类 号：R737.33[医药卫生—肿瘤]