下调ADAR1表达对人肺癌细胞增殖、迁移和上皮间质转化的影响

doi:10.13481/j.1671-587x.20200401

吉林大学学报(医学版) ›› 2020, Vol. 46 ›› Issue (04): 669-674.doi: 10.13481/j.1671-587x.20200401

• 基础研究 •

下调ADAR1表达对人肺癌细胞增殖、迁移和上皮间质转化的影响

周宁¹, 武睿², 马振凯³, 陈微微², 李雪琳², 宫凯凯², 杨丽娟², 代娟娟², 武艳²

1. 滨州医学院附属医院耳鼻咽喉头颈外科, 山东滨州 256600;
2. 滨州医学院附属医院肿瘤研究实验室, 山东滨州 256600;
3. 山东省滨州市人民医院神经外科, 山东滨州 256600

收稿日期:2019-12-10 发布日期:2020-08-20
通讯作者: 武艳,讲师(Tel:0543-3258276,E-mail:wuyan55@126.com);代娟娟,讲师(Tel:0543-3258276,E-mail:juanjdai@163.com) E-mail:wuyan55@126.com;juanjdai@163.com
作者简介:周宁(1986-),男,山东省滨州市人,医师,医学硕士,主要从事肿瘤发病机制方面的研究。
基金资助:
国家自然科学基金资助课题（81903102）；山东省科技厅重点研发计划项目资助课题（2019GSF107099）；山东省卫健委医药卫生科技发展计划项目资助课题（2017WS154）；滨州医学院科技计划项目资助课题（BY2019KJ04）；山东省科技厅自然科学基金资助课题（ZR2016HB55，ZR2018BB024，ZR2017PH028）

Effects of down-regulation of ADAR1 expression on proliferation, migration and epithelial-mesenchymal transition of human lung cancer cells

ZHOU Ning¹, WU Rui², MA Zhenkai³, CHEN Weiwei², LI Xuelin², GONG Kaikai², YANG Lijuan², DAI Juanjuan², WU Yan²

1. Deparment of Otolaryngology-Head and Neck Surgery, Affiliated Hospital, Binzhou Medical University, Binzhou 256603, China;
2. Cancer Research Institute, Affiliated Hospital, Binzhou Medical University, Binzhou 256603, China;
3. Department of Neurosurgery, Binzhou People's Hospital, Binzhou 256603, China

Received:2019-12-10 Published:2020-08-20

摘要/Abstract

摘要： 目的：探讨下调作用于RNA的腺苷酸脱氢酶1（ADAR1）对肺癌H1299和H520细胞增殖、迁移和上皮间质转化（EMT）的影响，并阐明其作用机制。方法：H1299和H520细胞系分为对照组（转染negative shRNA）和shADAR1组（转染shADAR1）。实时荧光定量PCR法和Western blotting法检测各组细胞中ADAR1 mRNA和蛋白表达水平，CCK-8法和克隆形成实验检测各组细胞增殖活性和克隆形成数，细胞划痕实验检测各组细胞划痕愈合率，Western blotting法检测各组细胞中E钙黏附蛋白（E-cadherin）和波形蛋白（vimentin）蛋白表达水平。结果：与对照组比较，shADAR1组的H1299和H520细胞中ADAR1 mRNA及蛋白表达水平均明显降低（P<0.05或P<0.01）。与对照组比较，shADAR1组H1299和H520细胞增殖活性、克隆形成数量和细胞划痕愈合率均明显降低（P<0.05或P<0.01），H1299和H520细胞中E-cadherin蛋白表达水平明显升高（P<0.01），vimentin蛋白表达水平明显降低（P<0.01）。结论：下调ADAR1表达能够抑制肺癌细胞的增殖、迁移及EMT。

关键词: 作用于RNA的腺苷酸脱氢酶1, 肺肿瘤, 细胞增殖, 细胞迁移, 上皮间质转化

Abstract: Objective: To investigate the effects of down-regulation of adenosine deaminase acting on RNA enzyme 1 (ADAR1) on the proliferation, migration and epithelial-mesenchymal transition (EMT) of the lung cancer H1299 and H520 cells, and to elucidate their mechanisms. Methods: The H1299 and H520 cells were divided into control group (transfected with negative shRNA) and shADAR1 group (transfected with shADAR1). Real-time fluorescence quantitative PCR and Western blotting methods were used to detect the expression levels of ADAR1 mRNA and protein, respectively. CCK8 method and colony information assay were used to detect the proliferation activities and the amounts of colony formation;scratch assay was used to detect the scratch healing rates of the cells in various groups; Western blotting method was performed to detect the expression levels of E-cadherin and vimentin proteins in the cells in various groups. Results: Compared with control group, the expression levels of ADAR1 mRNA and protein in the H1299 and H520 cells in shADAR1 group were significantly decreased(P<0.05).Compared with control group, the proliferation activity, the colony formation amount and the scratch healing rate of the H1299 and H520 cells in shADAR1 group were significantly decreased (P<0.05).Compared with control group, the expression levels of E-cadherin protein in the H1299 and H520 cells in shADAR1 group were significantly increased(P<0.01), and the expression levels of vimentin protein were decreased (P<0.01). Conclusion: Down-regulation of ADAR1 expression contributes to inhibiting the proliferation, migration, and EMT of lung cancer cells.

Key words: adenosine deaminase acting on RNA enzyme 1, lung neoplasms, cell proliferation, cell migration, epithelial-mesenchymal transition

中图分类号:

R734.2

周宁, 武睿, 马振凯, 陈微微, 李雪琳, 宫凯凯, 杨丽娟, 代娟娟, 武艳. 下调ADAR1表达对人肺癌细胞增殖、迁移和上皮间质转化的影响[J]. 吉林大学学报(医学版), 2020, 46(04): 669-674.

ZHOU Ning, WU Rui, MA Zhenkai, CHEN Weiwei, LI Xuelin, GONG Kaikai, YANG Lijuan, DAI Juanjuan, WU Yan. Effects of down-regulation of ADAR1 expression on proliferation, migration and epithelial-mesenchymal transition of human lung cancer cells[J]. Journal of Jilin University(Medicine Edition), 2020, 46(04): 669-674.

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下调ADAR1表达对人肺癌细胞增殖、迁移和上皮间质转化的影响

Effects of down-regulation of ADAR1 expression on proliferation, migration and epithelial-mesenchymal transition of human lung cancer cells

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