periostin促进Ishikawa子宫内膜癌肿瘤干细胞的增殖和侵袭  

periostin promotes proliferation and invasion of Ishikawa endometrial cancer stem cells

作　　者：苏萍萍 施琦蓉 陈少蓉[1] 张书铟 黄巧艺 Su Pingping;Shi Qirong;Chen Shaorong(Department of Obstetrics and Gynecology,The Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000)

机构地区：[1]福建医科大学附属第二医院妇产科,泉州362000

出　　处：《现代妇产科进展》2020年第6期409-415,共7页Progress in Obstetrics and Gynecology

基　　金：福建省自然科学基金资助项目(No:2017J01182)。

摘　　要：目的:探讨periostin对子宫内膜癌肿瘤干细胞增殖和侵袭的影响,及其可能作用机制。方法:收集100例手术切除的子宫内膜癌组织及距病灶3cm的癌旁组织,免疫组化、荧光定量PCR技术检测子宫内膜癌组织及癌旁组织中periostin的表达。流式细胞仪分选出子宫内膜癌细胞系Ishikawa CD133^+细胞并培养,免疫荧光检测Ishikawa肿瘤细胞球中CD133的表达情况。Western bolt法检测CD133^+Ishikawa和parental Ishikawa细胞中periostin的表达。构建针对periostin的shRNA表达载体转染Ishikawa CD133^+细胞和过表达periostin的载体转染亲本parental Ishikawa细胞。Western bolt法检测细胞中periostin蛋白表达水平,CCK-8和Transwell小室检测细胞的增殖和侵袭能力,Western bolt法检测Wnt信号通路蛋白(β-catenin、c-myc、CyclinD1、GSK3β)的表达。结果:periostin在子宫内膜癌组织的表达高于癌旁组织(P<0.01),并与子宫内膜癌的病理级别呈正相关(r=0.65,P<0.01)。子宫内膜癌肿瘤干细胞具有"干细胞"特性,能形成肿瘤球并表达干细胞标记物CD133。periostin在CD133^+Ishikawa细胞中的表达高于亲本parental Ishikawa细胞(P<0.01);相对于转染空载体组细胞,periostin shRNA能有效抑制Ishikawa CD133^+细胞的增殖和侵袭,且抑制Wnt信号通路蛋白(β-catenin、c-myc、CyclinD1、GSK3β)的表达(均P<0.01)。Parental Ishikawa细胞过表达periostin慢病毒能增强其增殖和侵袭能力,表现为Wnt/β-catenin信号通路被激活。结论:periostin通过激活Wnt/β-catenin信号通道促进子宫内膜癌肿瘤干细胞的增殖和侵袭。periostin可能成为临床治疗子宫内膜癌的新靶标。Objective:To investigate the effects of periostin on the proliferation and invasion of endometrial cancer stem cells and its possible mechanism.Methods:100 cases of surgically resected endometrial carcinoma tissues and 3 cm adjacent tissues were collected.Immunohistochemistry and real-time PCR were used to detect the expression of periostin in endometrial carcinoma tissues and adjacent tissues.Flow cytometry was used to isolate and culture the cells of Ishikawa CD133^+in endometrial cancer cells.The expression of CD133 in Ishikawa tumor cells was detected by immunofluorescence,Western bolt was used to detect the expression of CD133^+Ishikawa periostin and parental Ishikawa periostin.The shRNA expression vector for periostin was transfected into human endometrial cancer Ishikawa CD133^+cells.The parental Ishikawa cells were transfected with the overexpressing periostin vector.The expression level of periostin protein in all the group cells were detected by Western blot,the proliferation and invasion ability of the cells were detected by CCK-8 and Transwell chamber assay.The expression levels of Wnt signal related moleculesβ-catenin,c-myc,CyclinD1,GSK3βof all the group cells were detected by Western blot.Results:The expression of periostin in endometrial carcinoma tissues was higher than that in adjacent tissues(P<0.01)and positively correlated with the pathological grade of endometrial cancer(r=0.65,P<0.01).Endometrial cancer stem cells had"stem cell"characteristics,which can form tumor spheres and express stem cell marker CD133.periostin expression in CD133^+Ishikawa cells was higher than that in parental Ishikawa cells(P<0.01).periostin shRNA can effectively inhibit the proliferation and invasion of ECSCs and inhibit the expression of Wnt signaling pathway proteins(β-catenin,c-myc,CyclinD1,GSK3β)compared with transfected empty vector cells(P<0.01).In contrast,parental Ishikawa cells overexpressing the periostin lentiviral enhanced its proliferation and invasion,and the Wnt/β-catenin signaling pathway was ac

关 键 词：子宫内膜癌 肿瘤干细胞 PERIOSTIN 信号通路 增殖 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]