LINC00665通过靶向miR-138/AKT1促进Hela细胞增殖和抑制凋亡的机制研究  

LINC00665promotes Hela cell proliferation and inhibits apoptosis by targeting miR-138/AKT1

作　　者：刘晓娟[1] 谢双双 张晶[1] 康燕华[1] LIU Xiao-juan;XIE Shuang-shuang;ZHANG Jing(Department of Obstetrics and Gynecology,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075002,China)

机构地区：[1]河北北方学院附属第一医院妇产科,河北张家口075002

出　　处：《中国实验诊断学》2021年第5期750-755,共6页Chinese Journal of Laboratory Diagnosis

摘　　要：目的探究长链非编码RNA LINC00665通过靶向微小RNA(microRNA,miR)-138/AKT1促进Hela细胞增殖和抑制凋亡的机制。方法分析TCGA数据库中宫颈癌患者miR-138水平与生存之间的关系。通过双荧光素酶报告验证LINC00665靶向miR-138和miR-138靶向AKT1。将宫颈癌细胞系Hela分为4组:对照组、mimic组、mimic+LINC00665组和LINC00665组,通过质粒转染技术过表达miR-138和/或LINC00665。分别通过qPCR和Western blot检测RNA和蛋白的水平。分别通过CCK-8法和流式细胞术检测各组的细胞活力和凋亡率。通过荷瘤裸鼠实验在体内验证miR-138和LINC00665对肿瘤生长的影响。结果在TCGA数据库中,miR-138高表达的宫颈癌患者的生存率显高于miR-138低表达患者(P<0.05)。LINC00665直接靶向miR-138,miR-138直接靶向AKT1。Mimic组的miR-138水平显著高于对照组,LINC00665、AKT1mRNA和蛋白水平显著低于对照组(P<0.05)。LINC00665组miR-138水平显著低于对照组,LINC00665、AKT1mRNA和蛋白水平显著高于对照组(P<0.05)。mimic+LINC00665组的miR-138水平显著低于mimic组,LINC00665、AKT1mRNA和蛋白水平显著高于mimic组(P<0.05)。mimic组的细胞活力显著低于对照组,细胞凋亡率显著高于对照组(P<0.05),LINC00665组的细胞活力显著高于对照组,细胞凋亡率显著低于对照组(P<0.05)。mimic+LINC00665组的细胞活力显著高于mimic组,细胞凋亡率显著低于mimic组(P<0.05)。miR-138组荷瘤裸鼠的肿瘤体积和质量均显著低于对照组(P<0.05),miR-138+LINC00665组肿瘤体积和质量显著高于miR-138组(P<0.05)。结论miR-138靶向抑制AKT1的表达,LINC00665直接靶向miR-138,并且可通过靶向miR-138促进AKT1的表达。LINC00665通过靶向调控miR-138/AKT1促进Hela细胞的增殖并抑制凋亡。Objective To investigate the mechanism of long-chain non-coding RNA LINC00665in promoting Hela cell proliferation and inhibiting apoptosis by targeting microRNA(miR)-138/AKT1.Methods The relationship between miR-138levels and survival in patients with cervical cancer in the TCGA database was analyzed.It was verified by double luciferase report that LINC00665targets miR-138and miR-138targets AKT1.The cervical cancer cell line Hela was divided into four groups:control group,mimic group,mimic+LINC00665group and LINC00665group.MiR-138and/or LINC00665were overexpressed by plasmid transfection technology.RNA and protein levels were detected by qPCR and Western blot,respectively.Cell viability and apoptotic rate of each group were detected by CCK-8method and flow cytometry,respectively.The effects of miR-138and LINC00665on tumor growth were verified in vivo by tumor-bearing nude mice experiments.Results In the TCGA database,the survival rate of patients with high expression of miR-138in cervical cancer was significantly higher than that of patients with low expression of miR-138(P<0.05).LINC00665directly targets miR-138and miR-138directly targets AKT1.The miR-138levels in the Mimic group were significantly higher than those in the control group,and the levels of LINC00665,AKT1mRNA and protein were significantly lower than those in the control group(P<0.05).The levels of miR-138in LINC00665group were significantly lower than those in control group,and the levels of mRNA and protein of LINC00665,AKT1were significantly higher than those in control group(P<0.05).The miR-138levels in the mimic+LINC00665group were signifi cantly lower than those in the mimic group,and the levels of mRNA and protein of LINC00665,AKT1were significantly higher than those in the mimic group(P<0.05).The cell viability of the mimic group was significantly lower than that of the control group,and the apoptosis rate was significantly higher than that of the control group(P<0.05).The cell viability of the LINC00665group was significantly higher than t

关 键 词：宫颈癌 长链非编码RNA LINC00665 微小RNA-138 AKT1 

分 类 号：R737.33[医药卫生—肿瘤]