miR-92a通过调控NF-κB信号通路及EMT过程对宫颈癌细胞放疗敏感性的影响  

Effect of miR-92a on radiosensitivity of cervical cancer cells by regulating NF-κB signal pathway and EMT process

作　　者：戴刚毅[1] 严沁[1] 曾贵林[1] 杨雪梅[1] 黄远东[1] 何朗 DAI Gangyi;YAN Qin;ZENG Guilin;YANG Xuemei;HUANG Yuandong;HE Lang(Cancer Prevention and Treatment Institute of Chengdu,Department of Oncology,Chengdu Fifth People's Hospital,The Fifth Affiliated People's Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都市第五人民医院·成都中医药大学附属第五人民医院肿瘤科•成都市肿瘤防治所,四川成都611137

出　　处：《西部医学》2023年第5期639-647,共9页Medical Journal of West China

基　　金：成都市卫计委项目(2021144)。

摘　　要：目的探究miR-92a通过调控核因子-κB(NF-κB)信号通路及上皮间质转化(EMT)过程对宫颈癌细胞放疗敏感性的影响。方法收集2021年6月—2022年6月于本院行手术切除的50例宫颈癌组织标本及其癌旁正常组织标本,体外培养的细胞株包括正常人宫颈上皮HCvEpC细胞及宫颈癌细胞系Me 180、Caski、C-33A、Hela。qRT-PCR检测宫颈癌组织和细胞中miR-92a、NF-κB mRNA的表达;分析miR-92a表达与患者临床病理参数的关系;靶基因预测、双荧光素酶验证miR-92a对NF-κB的靶向调控作用;Pearson检验分析宫颈癌组织中miR-92a与NF-κB mRNA的相关性;将Hela细胞分为对照组、miR-NC组、miR-92a组、miR-92a inhibitor组、miR-92a inhibitor+pcDNA-NC组和miR-92a inhibitor+NF-κB组,采用脂质体瞬时转染法分别转染相应的质粒;MTT法、CCK-8、Transwell小室实验、划痕实验、流式细胞术分别检测各组细胞的增殖、侵袭、迁移、凋亡及细胞周期分布情况;Western blot分析各组细胞中NF-κB及EMT相关蛋白的表达。结果宫颈癌组织和细胞中miR-92a和NF-κB mRNA的表达均显著上调(P<0.05);miR-92a表达与患者的肿瘤分化程度、临床分期、淋巴结转移有明显的相关性(P<0.05);miR-92a靶向促进NF-κB表达;宫颈癌组织中miR-92a与NF-κB mRNA呈明显正相关(P<0.05);与对照组和miR-NC组相比,miR-92a组细胞OD值、单克隆形成数目、细胞侵袭数目、划痕愈合率、停留在S期的比例明显增加,NF-κB、N-cadherin、Vimentin蛋白表达水平明显升高,细胞凋亡率、停留在G0/G1期的比例、E-cadherin、α-catenin蛋白表达水平明显下降,miR-92a inhibitor组获得与上述相反的结果(P<0.05);与miR-92a inhibitor+pcDNA-NC组相比,miR-92a inhibitor+NF-κB组细胞OD值、单克隆形成数目、细胞侵袭数目、划痕愈合率、停留在S期的比例明显增加,NF-κB、N-cadherin、Vimentin蛋白表达水平明显升高,细胞凋亡率、停留在G0/G1期的比例、E-cadherin、α-catObjective To explore the effect of miR-92a on radiosensitivity of cervical cancer cells by regulating nuclear factor-κB(NF-κB)signal pathway and epithelial-mesenchymal transformation(EMT).Methods 50 cases of cervical cancer tissues and their adjacent normal tissues were collected from June 2021 to June 2022 in our hospital.The cell lines cultured in vitro included normal cervical epithelial HCvEpC cells and cervical cancer cell lines Me 180,Caski,Cmer 33A and Hela.The expression of miR-92a and NF-κB mRNA in cervical cancer tissues and cells was detected by qRT-PCR,and the relationship between miR-92a expression and clinicopathological parameters was analyzed.Target gene prediction and double luciferase were used to verify the targeted regulation of miR-92a on NF-κB.The correlation between miR-92a and NF-κB mRNA in cervical cancer was analyzed by Pearson test.Hela cells were divided into control group,miR-NC group,miR-92a group,miR-92a inhibitor group,miR-92a inhibitor+pcDNA-NC group and miR-92a inhibitor+NF-κB group,and the corresponding plasmids were transfected by liposome transient transfection,and the proliferation,invasion,migration,apoptosis and cell cycle distribution of cells were detected by MTT,CCK-8,Transwell chamber test,scratch test and flow cytometry,respectively.Western blot was used to analyze the expression of NF-κB and EMT-related proteins in each group.Results The expressions of miR-92a and NF-κB mRNA in cervical cancer tissues and cells were significantly up-regulated(P<0.05).The expression of miR-92a was significantly correlated with tumor differentiation,clinical stage and lymph node metastasis(P<0.05).MiR-92a targeted promoted the expression of NF-κB,and there was a significant positive correlation between miR-92a and NF-κB mRNA in cervical cancer(P<0.05).Compared with the control group and miR-NC group,the OD value,the number of monoclonal formation,the number of cell invasion,the rate of scratch healing and the proportion of staying in S phase were significantly increased,the ex

关 键 词：宫颈癌 miR-92a 核因子-ΚB 上皮间质转化 放疗敏感性 

分 类 号：R737.33[医药卫生—肿瘤]