ADAR1 p150和p110参与肝癌细胞增殖、迁移和侵袭  

Adenosine deaminase acting on RNA 1 p150 and p110 are involved in the proliferation,migration,and invasion of liver cancer cells

作　　者：巩雪 柳静 蹇文文 崔易红 涂增 Gong Xue;Liu Jing;Jian Wenwen;Cui Yihong;Tu Zeng(Department of Pathogen Biology,School of Basic Medical Sciences,Chongqing Medical University)

机构地区：[1]重庆医科大学基础医学院病原生物学教研室,重庆400016

出　　处：《重庆医科大学学报》2024年第7期820-829,共10页Journal of Chongqing Medical University

基　　金：国家自然科学基金青年科学基金资助项目(编号81501751);重庆市科技局自然科学基金面上资助项目(编号:cstc2021jcyj-msxmX0158)。

摘　　要：目的:研究腺苷脱氨酶1(adenosine deaminase acting on RNA 1,ADAR1)对肝细胞癌(liver hepatocellular carcinoma,LIHC)增殖、迁移与侵袭的影响。方法:利用基因表达谱交互式分析网站GEPIA检测ADAR1在LIHC中的表达水平和预后价值;提取人体肝癌标本的蛋白,通过Western blot实验检测ADAR1的表达情况。将ADAR1 p150和p110过表达质粒和靶向ADAR1 p150和p110的小干扰RNA分别转染进HepG2和Huh7细胞,利用Western blot和qPCR检测转染后细胞的ADAR1表达情况。通过CCK-8实验检测ADAR1对肝癌细胞增殖能力的影响。通过细胞划痕实验和Transwell实验检测ADAR1对肝癌细胞迁移和侵袭的影响。最后,在DAVID在线数据库中分析与ADAR1相关的信号通路,利用Western blot实验检测信号通路中相关蛋白的表达。结果:肝癌组织中ADAR1蛋白质水平明显高于癌旁组织,这与生物信息学的预测一致,且高表达的ADAR1与肝癌的不良预后相关;通过CCK-8实验,本研究发现过表达ADAR1可以促进肝癌细胞的增殖能力(P<0.05),敲低ADAR1使肝癌细胞增殖能力下降(P<0.05)。通过划痕和Transwell实验,本研究发现ADAR1的过表达促进了肝癌细胞的迁移和侵袭(P<0.05),敲低ADAR1后,肝癌细胞迁移和侵袭能力受到抑制(P<0.05)。DAVID在线分析结果显示,ADAR1主要富集在Wnt信号通路中。Western blot结果提示,过表达ADAR1抑制GSK3β表达和β-catenin的磷酸化水平,敲低ADAR1后GSK3β表达和β-catenin的磷酸化水平升高。结论:本研究结果表明,ADAR1在肝癌中处于高表达水平,可激活Wnt/β-catenin信号通路,从而促进肝癌的增殖、迁移和侵袭。Objective:To investigate the effect of adenosine deaminase acting on RNA 1(ADAR1)on the proliferation,migration,and invasion of liver hepatocellular carcinoma(LIHC)cells.Methods:The gene expression profiling interactive analysis website GEPIA was used to obtain the expression level and prognostic value of ADAR1 in LIHC;proteins were extracted from human hepatocellular carcinoma specimens,and Western blot was used to measure the expression level of ADAR1.HepG2 and Huh7 cells were transfected with ADAR1 p150 and p110 overexpression plasmids or small interfering RNAs targeting both ADAR1 p150 and p110,and Western blot and qPCR were used to measure the expression of ADAR1 in cells after transfection.CCK-8 assay was used to observe the effect of ADAR1 on the proliferative ability of hepatocellular carcinoma cells,and scratch assay and Transwell assay were used to observe the effect of ADAR1 on the migration and invasion of LIHC cells.Finally,the DAVID online database was used to analyze the signaling pathways related to ADAR1,and Western blot was used to measure the expression of proteins associated with the signaling pathways.Results:The protein expression level of ADAR1 in hepatocellular carcinoma tissue was significantly higher than that in paracancerous tissue,which was consistent with the prediction based on bioinfor⁃matics,and the high expression of ADAR1 was associated with the poor prognosis of hepatocellular carcinoma.CCK-8 assay showed that overexpression of ADAR1 promoted the proliferation ability of hepatocellular carcinoma cells(P<0.05),and knockdown of ADAR1 reduced the proliferation ability of hepatocellular carcinoma cells(P<0.05).Scratch assay and Transwell assay showed that overexpression of ADAR1 significantly promoted the migration and invasion of hepatocellular carcinoma cells(P<0.05),and knock⁃down of ADAR1 inhibited the migration and invasion abilities of hepatocellular carcinoma cells(P<0.05).The results of DAVID online analysis showed that ADAR1 was mainly enriched in the Wnt signaling pat

关 键 词：腺苷脱氨酶1 肝癌 增殖 迁移侵袭 WNT/Β-CATENIN信号通路 

分 类 号：R735.7[医药卫生—肿瘤]