lncRNA HNF1A-AS1通过miR-320d/SOX4轴调节宫颈癌的发展  

LncRNA HNF1A-AS1 regulates the development of cervical cancer through miR-320d/SOX4 axis

作　　者：谢锦霞 刘晶晶[2] XIE Jinxia;LIU Jingjing(Department of Reproductive Medicine,Xi'an Fourth Hospital,Xi'an Shaanxi 710100,P.R.China;Department of Obstetrics and Gynecology,Xi'an Fourth Hospital,Xi'an Shaanxi 710100,P.R.China)

机构地区：[1]西安市第四医院生殖科,陕西西安710100 [2]西安市第四医院妇产科,陕西西安710100

出　　处：《中国计划生育和妇产科》2021年第6期45-50,共6页Chinese Journal of Family Planning & Gynecotokology

基　　金：陕西省重点研发计划项目(项目编号:2019 SF-210)。

摘　　要：目的探讨长链非编码RNA HNF1A-AS1(long noncoding RNA HNF1A-AS1,lncRNA HNF1A-AS1)对宫颈癌细胞增殖、凋亡和上皮细胞-间充质转化(epithelial-mesenchymal transition,EMT)的影响及机制研究。方法采用实时荧光定量PCR(quantitative real-time PCR,RT-qPCR)检测微小RNA 320d(microRNA-320d,miR-320d)在人宫颈癌细胞C33A和人宫颈上皮永生化细胞株H8细胞中的表达水平,分别用四甲基偶氮唑蓝(methylthiazolyl-tetrazolium,MTT)比色法、流式细胞仪和Western blot检测miR-320d对C33A细胞增殖、凋亡和EMT的影响。采用miRanda和双荧光素酶报告基因实验分析miR-320d与HNF1A-AS1作用的靶点,检测HNF1A-AS1通过miR-320d对C33A细胞增殖、凋亡与EMT的影响。通过TargetScan和双荧光素酶报告基因实验分析miR-320d与SOX4之间的关系,检测SOX4对C33A细胞增殖、凋亡和EMT的影响,采用RT-qPCR和Western blot分析HNF1A-AS1及miR-320d对SOX4表达的影响。结果在C33A细胞中,miR-320d表达明显降低,HNF1A-AS1和SOX4表达明显升高,上调miR-320d抑制C33A细胞的增殖和EMT,促进癌细胞凋亡。HNF1A-AS13'UTR靶向miR-320d,敲低HNF1A-AS1抑制了C33A细胞增殖和EMT,促进癌细胞凋亡。下调HNF1A-AS1和miR-320d表达能部分逆转miR-320d下调对C33A细胞增殖、凋亡和EMT的影响。miR-320d和SOX4具有靶向作用,敲低SOX4抑制了C33A细胞增殖和EMT,促进了癌细胞凋亡。下调HNF1A-AS1使SOX4表达量下降,miR-320d表达量升高,而且HNF1A-AS1能够通过miR-320d调控SOX4的表达。结论lncRNA HNF1A-AS1通过miR-320d/SOX4轴调节宫颈癌的发展。Objective To investigate the effect of lncRNA HNF1 A-AS1 on proliferation,apoptosis and epithelial-mesenchymal transition(EMT)of cervical cancer cells and the underlying mechanism.Methods RT-qPCR was used to detect the expression of miR-320 d in C33 A and H8 cells.The effects of miR-320 d on the proliferation,apoptosis of C33 A cells and EMT were analyzed by MTT experiments,flow cytometry and Western blot experiments respectively.The miRanda and Dual luciferase reporter gene assay were used to analyze the correlation between HNF1 A-AS1 and miR-320 d.The effects of HNF1 A-AS1 on the proliferation,apoptosis and EMT of C33 A cells through miR-320 d were analyzed.The TargetScan and Dual luciferase reporter gene assay were used to analyze the correlation between miR-320 d and SOX4.The effects of SOX4 on the proliferation,apoptosis of C33 A cells and EMT were analyzed.The effects of HNF1 A-AS1 and miR-320 d on SOX4 expression were analyzed by RT-qPCR and Western blot.Results In C33 A cells,the expression of miR-320 d was significantly decreased,and the expressions of HNF1 A-AS1 and SOX4 were significantly increased.Upregulation of miR-320 d inhibited the proliferation of C33 A cells and EMT and promoted the apoptosis of cancer cells.HNF1 A-AS13’UTR targeted miR-320 d.The down-regulation of HNF1 A-AS1 inhibited the proliferation of C33 A cells and EMT and promoted the apoptosis of cancer cells.The down-regulation of the HNF1 A-AS1 and miR-320 d can partially reverse the effects of miR-320 d down-regulation on the proliferation,apoptosis of C33 A cells and EMT.SOX43’UTR targeted miR-320 d.The down-regulation of SOX4 inhibited C33 A cells proliferation,EMT and promoted cells apoptosis.The down-regulation of the HNF1 A-AS1 decreased the expression of SOX4 and increased the expression of miR-320 d,and HNF1 A-AS1 can regulate the expression of SOX4 through miR-320 d.Conclusion LncRNA HNF1 A-AS1 regulates the development of cervical cancer through miR-320 d/SOX4 axis.

关 键 词：HNF1A-AS1 miR-320d SOX4 宫颈癌 增殖 EMT 

分 类 号：R737.33[医药卫生—肿瘤]