MEX3A在非小细胞肺癌中的表达与功能研究

朱贝; 郭小朋; 董天祺; 肖春杰

doi:10.3969/j.issn.1000-8179.2020.15.571

MEX3A在非小细胞肺癌中的表达与功能研究

doi: 10.3969/j.issn.1000-8179.2020.15.571

朱贝^①,,
郭小朋^②,
董天祺^②,
肖春杰^②, ,

①.
黄石市中医医院(市传染病医院)湖北中医药大学黄石临床学院医学检验科(黄石市 435000)
②.
云南大学医学院

基金项目:

国家自然科学基金项目 81860499

详细信息

作者简介:
朱贝专业方向为肿瘤遗传与分子机理的相关研究。E-mail:2671668602@qq.com

通讯作者:
肖春杰 chjxiao@ynu.edu.cn

计量
- 文章访问数: 143
- HTML全文浏览量: 4
- PDF下载量: 22
- 被引次数: 0
出版历程
- 收稿日期: 2020-05-08
- 刊出日期: 2020-12-26

Expression level and function of MEX3A in non-small cell lung cancer

Bei Zhu^{①
,},
Xiaopeng Guo^②,
Tianqi Dong^②,
Chunjie Xiao^{②
, ,}

①.
Medicine Clinical Laboratory, Huangshi Clinical College of Hubei University of Chinese Medicine, Huangshi Hospital of TCM(Infectious Disease Hospital), Huangshi 435000, China
②.
Yunnan University School of Medicine, Kunming 650091, China

Funds:

National Natural Science Foundation of China 81860499

More Information

Corresponding author: Chunjie Xiao; E-mail: chjxiao@ynu.edu.cn

摘要

摘要: 目的探究MEX3A基因在非小细胞肺癌（non-small cell lung cancer，NSCLC）中的表达水平及沉默MEX3A基因后对NSCLC增殖、侵袭、迁移和凋亡及周期的影响。方法通过mRNA转录组分析及癌症基因图谱（The Cancer Genome Atlas，TCGA）数据库信息分析，筛选出在NSCLC中显著高表达的MEX3A基因。通过qRT-PCR检测MEX3A在4种常见NSCLC细胞系中的mRNA表达水平，发现MEX3A在A549和NCI-H292中显著高表达。利用RNA干扰技术（RNA interference，RNAi）沉默A549和NCI-H292中MEX3A的表达，CCK8及Transwell试验检测沉默MEX3A表达后对NSCLC增殖、侵袭及迁移的影响，流式细胞术分析沉默MEX3A表达后对A549和NCI-H292细胞周期及凋亡的影响。结果 MEX3A在A549和NCI-H292中高表达。沉默MEX3A后NSCLC增殖、侵袭及迁移被显著抑制，细胞周期阻滞于G2/M期，促进细胞凋亡。结论 MEX3A作为NSCLC的促癌基因，参与了NSCLC的生长及增殖过程。
- MEX3A /
- 非小细胞肺癌 /
- 基因沉默 /
- 促癌基因
Abstract: Objective To investigate the expression of MEX3A in non-small cell lung cancer (NSCLC) cells and the effects of MEX3A knockout on cell cycle, proliferation, invasion, migration, and apoptosis. Methods We screened out MEX3A, which was significantly highly expressed, by mRNA microarray and The Cancer Genome Atlas (TCGA) database information analysis. Quantitative real-time polymerase chain reaction (qRT- PCR) was used to detect the expression level of MEX3A in NSCLC cells. A549 and NCI- H292 cells showed high expression of MEX3A. RNA interference (RNAi) method was used to silence MEX3A in A549 and NCI- H292 cells. Cell counting kit-8 (CCK8) and Transwell assays were used to determine the effects of MEX3A knockout on proliferation, invasion, and migration in NSCLC cells. Flow cytometry was used to analyze the effects of MEX3A knockout on cell cycle and apoptosis. Results A549 and NCI-H292 cells showed high expression of MEX3A. After silencing MEX3A, the proliferation, invasion, and migration of NSCLC cells were significantly decreased, while the cell cycle was blocked at G2/M phase and its apoptotic ability was weakened. Conclusions MEX3A may play an important role as an oncogene in the growth and proliferation of NSCLC cells.
- muscle excess-3 A (MEX3A) /
- non-small cell lung cancer (NSCLC) /
- gene silencing /
- oncogene

HTML全文

图 1 MEX3A在NSCLC中高表达

A：TCGA数据库数据统计分析显示与正常组织样本（n=20）相比，MEX3A mRNA在肺腺癌组织（n=226）中显著高表达；B：RT-qPCR结果显示A549和NCIH292的MEX3A mRNA表达水平显著高于正常癌旁细胞BEAS-2B；C：qRT-PCR结果显示沉默MEX3A基因后A549和NCI-H292的mRNA表达水平显著降低；***：P＜0.001

下载: 全尺寸图片幻灯片

图 2 沉默MEX3A基因后抑制NSCLC的增殖

下载: 全尺寸图片幻灯片

图 3 沉默MEX3A基因后抑制NSCLC的侵袭及迁移

***：P＜0.001

下载: 全尺寸图片幻灯片

图 4 沉默MEX3A基因后诱导NSCLC细胞凋亡

*：P＜0.05；***：P＜0.001

下载: 全尺寸图片幻灯片

图 5 沉默MEX3A基因后阻滞NSCLC细胞周期

***：P＜0.001

下载: 全尺寸图片幻灯片

表 1 引物序列

表 2 新鲜组织样本临床病理资料

表 3 mRNA转录组芯片筛选出在NSCLC中上调的mRNAs

参考文献(17)

[1]	Wen C, Dehnel T. China wrestles with lung cancer[J]. Lancet Oncol, 2011, 12(1):15. doi: 10.1016/S1470-2045(10)70303-X
[2]	Travis WD, Brambilla E, Nicholson AG, et al. The 2015 World Health Organization Classification of lung tumors:impact of genetic, clinical and radiologic advances since the 2004 classification[J]. J Thorac Oncol, 2015, 10(9):1243-1260. doi: 10.1097/JTO.0000000000000630
[3]	Buchet-poyau K, Coerchet J, Le hir H, et al. Identification and characterization of human Mex-3 proteins, a novel family of evolutionarily conserved RNA-binding proteins differentially localized to processing bodies[J]. Nucleic Acids Res, 2007, 35(4):1289-1300. doi: 10.1093/nar/gkm016
[4]	Valverde R, Edwards L, Regan L. Structure and function of KH domains[J]. FEBS J, 2008, 275(11):2712-2726. doi: 10.1111/j.1742-4658.2008.06411.x
[5]	Cano F, Bye H, Duncan LM, et al. The RNA-binding E3 ubiquitin ligase MEX-3C links ubiquitination with MHC-I mRNA degradation[J]. EMBO J, 2012, 31(17):3596-3606. doi: 10.1038/emboj.2012.218
[6]	Jiang H, Zhang X, Luo J, et al. Knockdown of hMex-3A by small RNA interference suppresses cell proliferation and migration in human gastric cancer cells[J]. Mol Med Rep, 2012, 6(3):575-580. http://cn.bing.com/academic/profile?id=c8c0a41133e1453e029ceb4d2f40e5e9&encoded=0&v=paper_preview&mkt=zh-cn
[7]	Krepischi ACV, Maschietto M, Ferreira EN, et al. Genomic imbalances pinpoint potential oncogenes and tumor suppressors in Wilms tumors[J]. Mol Cytogenet, 2016, 9:20. doi: 10.1186/s13039-016-0227-y
[8]	Shi JW, Huang Y. Mex3a expression and survival analysis of bladder urothelial carcinoma[J]. Oncotarget, 2017, 8(33):54764-54774. doi: 10.18632/oncotarget.18399
[9]	Yang DJY, Li Y, Fang X. Clinical characteristics and prognostic value of MEX3A mRNA in liver cancer[J]. PeerJ, 2020, 8:e8252. doi: 10.7717/peerj.8252
[10]	徐嵩, 赵世康, 任凡, 等.非小细胞肺癌新辅助靶向及免疫治疗研究进展和展望[J].中国肿瘤临床, 2020, 47(6):299-303. doi: 10.3969/j.issn.1000-8179.2020.06.151
[11]	Pao W, Girard N. New driver mutations in non-small cell lung cancer[J]. Lancet Oncol, 2011, 12(2):175-180. doi: 10.1016/S1470-2045(10)70087-5
[12]	Draper BW, Mello CC, Bowerman B, et al. MEX-3 is a KH domain protein that regulates blastomere identity in early C. elegans embryos[J]. Cell, 1996, 87(2):205-216. doi: 10.1016/S0092-8674(00)81339-2
[13]	Sheth U, Parker R. Decapping and decay of messenger RNA occur in cytoplasmic processing bodies[J]. Science, 2003, 300(5620):805-808. doi: 10.1126/science.1082320
[14]	Teixeira D, Sheth U, Valencin-Sanchez MA, et al. Processing bodies require RNA for assembly and contain nontranslating mRNAs[J]. RNA, 2005, 11(4):371-382. doi: 10.1261/rna.7258505
[15]	陈云云, 房超, 于秋爽, 等.MEX3A在人膀胱癌组织细胞中的表达及分析[J].生物医学工程与临床, 2018, 22(1):82-86. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=swyxgcylc201801020
[16]	房超, 尹晶, 于秋爽, 等.RNA干扰MEX3A基因对膀胱癌细胞增殖和凋亡的影响[J].解剖学报, 2017, 48(6):688-692. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=jpxb201706011
[17]	Pereira B, Sousa S, Barros R, et al. CDX2 regulation by the RNA-binding protein MEX3A:impact on intestinal differentiation and stemness[J]. Nucleic Acids Res, 2013, 41(7):3986-3999. doi: 10.1093/nar/gkt087