摘要
目的探讨长链非编码RNA(lncRNA)白介素增强因子3反义1(ILF3-AS1)对宫颈癌细胞增殖、迁移和侵袭的影响和可能机制。方法实时荧光定量PCR(RT-qPCR)检测宫颈癌组织、癌旁组织中ILF3-AS1和miR-204的表达水平。将ILF3-AS1小干扰RNA、miR-204模拟物分别转染SiHa细胞,采用四甲基偶氮唑蓝(MTT)法和集落形成实验检测细胞增殖,流式细胞术检测细胞周期分布,Transwell实验检测细胞迁移和侵袭,蛋白质印记(Western blot)检测白介素6受体(IL-6R)表达。双荧光素酶报告实验验证ILF3-AS1与miR-204、miR-204与IL-6R的调控关系。结果与癌旁组织比较,宫颈癌组织中ILF3-AS1表达显著升高,miR-204表达显著降低(P<0.05)。干扰ILF3-AS1表达后SiHa细胞存活率、克隆形成数、迁移和侵袭细胞数、S期细胞比例、IL-6R蛋白表达显著降低,G0-G1期细胞比例、miR-204表达显著升高(P<0.05)。过表达miR-204后SiHa细胞存活率、克隆形成数、迁移和侵袭细胞数、S期细胞比例、IL-6R蛋白表达显著降低,G0-G1期细胞比例显著升高(P<0.05)。抑制miR-204表达可逆转干扰ILF3-AS1对SiHa细胞增殖、周期分布、迁移和侵袭的影响(P<0.05)。结论干扰ILF3-AS1可能通过上调miR-204/IL-6R途径来抑制宫颈癌细胞SiHa的增殖、迁移和侵袭。
This study was designed to investigate the effect and possible mechanism of long-chain non-coding RNA(lncRNA)interleukin-enhancing factor 3 antisense 1(ILF3-AS1)in the proliferation,migration and invasion of cervical cancer cells.Real-time quantitative PCR(RT-qPCR)was used to detect the expression levels of ILF3-AS1 and miR-204 in cervical cancer tissues and adjacent tissues.ILF3-AS1 small interfering RNA and miR-204 mimics were transfected into SiHa cells,respectively.Then cell proliferation was detected by methyl thiazolyl tetrazolium(MTT)method and colony formation experiment;flow cytometry was used to detect cycle distribution;Transwell experiment was used to detect cell migration and invasion;Western blot was used to detect the expression of interleukin 6 receptor(IL-6R)protein.The targeted relationships of ILF3-AS1 with miR-204,and miR-204 with IL-6R were verified by dual-luciferase reporter assay.Data showed that ILF3-AS1 expression in cervical cancer tissue was significantly increased,while miR-204 expression was significantly reduced(P<0.05),as compared to adjacent tissues.ILF3-AS1 interference could reduce the survival rate of SiHa cells,the number of clone formation,the number of migrating and invasive cells,the proportion of S-phase cells,and the expression of IL-6R protein significantly,while up-regulate the proportion of G0-G1 phase cells and the expression of miR-204 significantly(P<0.05).miR-204 overexpression could also reduce the survival rate of SiHa cells,the number of clone formation,the number of migrating and invasive cells,the proportion of S-phase cells,the expression of IL-6R protein significantly,but up-regulate the proportion of G0-G1 phase cells significantly(P<0.05).Inhibiting the expression of miR-204 can reverse ILF3-AS1 interference-induced effect on the proliferation,cycle distribution,migration and invasion of SiHa cells(P<0.05).In conclusion,ILF3-AS1 interference may inhibit the proliferation,migration and invasion of cervical cancer cell SiHa by up-regulating miR-204/IL-6R pathway.
作者
罗健玮
黄泓轲
胡艳丽
LUO Jianwei;HUANG Hongke;HU Yanli(Medical Department of Leshan Vocational and Technical College,Leshan 614000,China;Department of Pharmacy,Leshan Vocational and Technical College,Leshan 614000,China;Department of Gynecology and Obstetrics,Qianjiang Central Hospital,Chongqing 409099,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2021年第8期710-718,共9页
Immunological Journal
基金
乐山市科技局课题(20SZD004)。
