miR-760在宫颈鳞状细胞癌组织和细胞中的表达及其对SiHa细胞恶性生物学行为的作用  

Expression of miR-760 in cervical squamous cell carcinoma tissues and cells and its effect on malignant biological behaviors of SiHa cells

作　　者：邓振宇 何传凤 牛占杰[2] DENG Zhenyu;HE Chuanfeng;NIU Zhanjie(Department of Obstetrics and Gynecology,Liaocheng Maternal and Child Health Hospital,Liaocheng 252000,Shandong,China;Department of Obstetrics and Gynecology,Liaocheng People's Hospital,Liaocheng 252000,Shandong,China)

机构地区：[1]山东省聊城市妇幼保健院妇产科,山东聊城252000 [2]山东省聊城市人民医院妇产科,山东聊城252000

出　　处：《中国肿瘤生物治疗杂志》2019年第10期1120-1127,共8页Chinese Journal of Cancer Biotherapy

基　　金：山东省医药卫生科技发展计划资助项目（No.2016WS0583）~~

摘　　要：目的:探讨微小RNA(miR)-760在人宫颈鳞状细胞癌(CSCC)组织和细胞中的表达及其对SiHa细胞增殖、凋亡、侵袭、迁移、EMT影响的分子机制。方法:选用2015年4月至2018年8月山东省聊城市妇幼保健院妇科手术切除、经病理确诊的80例CSCC组织及相应的癌旁组织标本和40例子宫肌瘤切除术获取的正常宫颈组织标本,应用qPCR检测CSCC组织、癌旁组织和正常宫颈组织、人CSCC细胞系SiHa、HCC94和人宫颈鳞状上皮永生化细胞株H8中miR-760的表达水平,分析miR-760表达与CSCC患者临床病理特征的相关性。用脂质体转染法,将miR-760mimics和NC-mimics质粒分别转染至SiHa细胞,用qPCR检测SiHa细胞中miR-760的表达,用CCK-8实验和流式细胞术分别检测SiHa细胞增殖能力和凋亡水平,用Transwell实验检测SiHa细胞的侵袭和迁移能力,WB检测SiHa细胞EMT相关蛋白上皮型钙黏蛋白(E-cadherin)、波形蛋白(vimentin)和神经型钙黏蛋白(N-cadherin)的表达变化。用生物学信息法预测FOXA1与miR-760的靶向关系,用双荧光素酶报告基因验证miR-760对FOXA1的直接靶向调控作用。结果:CSCC组织中miR-760表达明显低于癌旁组织及正常宫颈组织(均P<0.01),miR-760表达与患者淋巴结转移和临床分期密切相关(均P<0.01)。SiHa、HCC94细胞中miR-760的表达明显低于H8细胞(均P<0.01)。通过转染miR-760mimics上调miR-760表达,能够显著抑制SiHa细胞的增殖能力和侵袭、迁移能力(均P<0.01)、促进其凋亡(P<0.01),并上调Ecadherin的表达(P<0.01)、下调vimentin和N-cadherin的表达(均P<0.01)。FOXA1是miR-760的直接靶基因(P<0.01),上调miR-760表达显著抑制SiHa细胞中FOXA1mRNA和蛋白的表达(均P<0.05)。结论:在CSCC组织和细胞中miR-760低表达,其通过靶向作用FOXA1基因调控SiHa细胞的增殖、侵袭、迁移并促进凋亡,同时影响癌细胞的EMT进程。Objective:To investigate the expression of microRNA(miR)-760 in human cervical squamous cell carcinoma(CSCC)tissues and cells,and it’s effects on the proliferation,apoptosis,invasion,migration and epithelial-mesenchymal transition(EMT)of SiHa cells,as well as its molecular mechanism.Methods:Eighty pairs of CSCC cancerous and corresponding para-cancerous tissue specimens which were pathologically confirmed and 40 cases of normal cervical tissue specimens obtained by myomectomy in Liaocheng Maternal and Child Health Hospital of Shandong Province from April 2015 to August 2018 were selected.The expression of miR-760 in CSCC tissues,para-cancerous tissues and normal cervical tissues,human CSCC cell lines(SiHa,HCC94)and human cervical squamous epithelial immortalized H8 cells were detected by qPCR,and the relationship between miR-760 and clinicopathological characteristics of CSCC patients was analyzed.miR-760 mimics and NC-mimics plasmids were transfected into SiHa cells by liposome transfection.The expression of miR-760 in SiHa cells was detected by qPCR,the proliferation activity and apoptosis rate were detected by CCK-8 test and flow cytometry,respectively.The invasion and migration of SiHa cells were detected by Transwell assay.The expres-sions of EMT-related proteins,such as E-cadherin,vimentin and N-cadherin,in SiHa cells were detected by WB.Bioinformatics was used to predict the targeting relationship between FOXA1 and miR-760,and double luciferase reporter gene assay was used to verify the direct regulation of miR-760 on FOXA1.Results:The expression of miR-760 in CSCC tissues was significantly lower than that in para-cancerous tissues and normal cervical tissues(all P<0.01),and the expression of miR-760 was closely related to lymphnode metastasis and clinical stage(all P<0.01).The expression of miR-760 in SiHa and HCC94 cells was significantly lower than that in H8 cells(all P<0.01).Up-regulation of the expression of miR-760 could significantly inhibit the proliferation,invasion and migration of SiHa cells(

关 键 词：宫颈鳞状细胞癌 SIHA细胞 miR-760 FOXA1 增殖 侵袭 迁移 凋亡 

分 类 号：R737.33[医药卫生—肿瘤] R730.2[医药卫生—临床医学]