lncRNA MEG3通过miR-9-5p/SOCS5轴对宫颈癌细胞恶性生物学行为的影响  

Effect of lncRNA MEG3 on the malignant biological behaviors of cervical cancer cells via miR-9-5p/SOCS5 axis

作　　者：张艳[1] 李封[2] 徐婉 张莉 段亚亭 余作琼 ZHANG Yan;LI Feng;XU Wan;ZHANG Li;DUAN Yating;YU Zuoqiong(Department of Gynaecology,Chongqing Hospital of Traditional Chinese Medicine,Chongqing 400011,China;Department of Oncology,Chongqing Hospital of Traditional Chinese Medicine,Chongqing 400011,China;7th Medical Center,General Hospital of PLA,Beijing 100039,China)

机构地区：[1]重庆市中医院妇科,重庆400011 [2]重庆市中医院肿瘤科,重庆400011 [3]解放军总医院第七医学中心,北京100039

出　　处：《中国肿瘤生物治疗杂志》2020年第7期725-734,共10页Chinese Journal of Cancer Biotherapy

基　　金：国家重点研发计划资助项目(No.2018YFC1704104)。

摘　　要：目的:探讨lncRNA母源性印记基因3(maternal imprinting gene 3,MEG3)通过miR-9-5p/SOCS5轴对宫颈癌细胞增殖、迁移、侵袭和上皮间质转化(epithelial-mesenchymal transition,EMT)的调控作用。方法:收集2017年1月至2019年6月重庆市中医院手术切除的20例宫颈癌患者的癌及癌旁组织标本;利用脂质体转染技术分别将pcDNA3.1-MEG3、si-MEG3、miR-9-5p mimics、miR-9-5p inhibitor及其对照质粒等转染进宫颈癌HeLa和SiHa细胞,构建过表达和沉默细胞模型。用qPCR检测宫颈癌组织及细胞模型中MEG3、miR-9-5p和SOCS5表达水平,用CCK-8法、Transwell小室法检测细胞的增殖、迁移和侵袭能力,用细胞免疫荧光实验检测细胞中E-cadherin和vimentin表达水平。通过在线生物信息学TargetScan数据库预测靶基因,用双荧光素酶报告基因实验验证miR-9-5p分别与MEG3和SOCS5的靶向关系。结果:分别与癌旁组织和宫颈上皮HcerEpic细胞比较,宫颈癌组织和细胞系中MEG3和SOCS5表达显著下调、miR-9-5p表达显著上调(均P<0.01)。TargetScan数据库分析和双荧光素酶报告基因实验证实miR-9-5p与MEG3或SOCS5存在靶向关系。MEG3和SOCS5显著抑制宫颈癌细胞的增殖、迁移与侵袭能力(均P<0.01),miR-9-5p显著提高细胞的增殖、迁移与侵袭能力(均P<0.01)。MEG3和SOCS5促进E-cadherin表达、抑制vimentin表达;miR-9-5p抑制E-cadherin表达、促进vimentin表达(P<0.05或P<0.01)。结论:lncRNA MEG3通过miR-9-5p/SOCS5分子轴调控宫颈癌细胞的增殖、迁移、侵袭与EMT进程。Objective:To explore the regulatory effect of lncRNA maternal imprinting gene 3(MEG3)on proliferation,migration,invasion and epithelial-mesenchymal transition(EMT)of cervical cancer cells via miR-9-5p/SOCS5 axis.Methods:A total of 20 pairs of cancer and para-cancerous tissue specimens resected from cervical cancer patients in Chongqing Hospital of Traditional Chinese Medicine from January 2017 to June 2019 were collected for this study.Using liposome transfection technology,pcDNA3.1-MEG3,si-MEG3,miR-9-5p mimics,miR-9-5p inhibitor and their control plasmids were transfected into cervical cancer HeLa and SiHa cells respectively to construct overexpression and silence cell model.qPCR was used to detect the expression levels of MEG3,miR-9-5p and SOCS5 in cervical cancer tissues and cell lines.CCK-8 method and Transwell chamber method were used to detect cell proliferation,migration and invasion ability.The expression levels of E-cadherin and vimentin in cells were detected by cellular immunofluorescence experiments.Target genes were predicted through the Online Bioinformatics TargetScan database.Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-9-5p and MEG3,SOCS5,respectively.Results:Compared with para-cancerous tissues and cervical epithelial HcerEpic cells,the expressions of MEG3 and SOCS5 were significantly down-regulated and the expression of miR-9-5p was significantly up-regulated in cervical cancer tissues and cell lines(all P<0.01).TargetScan database analysis and Dual luciferase reporter gene assay confirmed the targeting relationship between miR-9-5p and MEG3 or SOCS5.MEG3 and SOCS5 significantly inhibited while miR-9-5p significantly promoted cell proliferation,migration and invasion ability(all P<0.01).MEG3 and SOCS5 promoted E-cadherin expression and inhibited vimentin expression,while miR-9-5p inhibited E-cadherin expression and promoted vimentin expression(P<0.05 or P<0.01).Conclusion:lncRNA MEG3 regulates proliferation,migration,invasion and EMT of cervical

关 键 词：lncRNA母源性印记基因3 miR-9-5p/SOCS5轴 宫颈癌 HELA细胞 SIHA细胞 增殖 迁移 侵袭 上皮间质转化 

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