摘要
目的:探究长链非编码RNA(lncRNA)NORAD调控肺腺癌(LUAD)发生发展及其可能的作用机制。方法:RT-PCR检测19例肺腺癌与邻近正常组织NORAD的表达,分析NORAD表达量与临床病理分期关系;RT-PCR检测NORAD在A549和NCI-H1299细胞表达情况,si-NORAD干扰沉默A549和NCI-H1299细胞,检测si-NORAD对细胞增殖、迁移和侵袭能力的影响;Western Blot检测MMP9、MMP2、Vimentin、N-Cadherin及E-Cadherin表达情况;基于TCGA数据库提取LUAD转录组数据行加权基因共表达网络分析(WGCNA),进一步对促进LUAD可能的信号通路进行基因集富集分析(GSEA)NORAD,再行Western Blot验证。结果:NORAD在肺腺癌组织中高表达,且更易发生淋巴结转移;NORAD在A549和NCI-H1299细胞明显过表达,敲低NORAD能明显降低A549和NCI-H1299细胞增殖、迁移和侵袭能力;敲低NORAD,MMP9、MMP2、Vimentin、N-Cadherin表达上调,E-Cadherin表达下调;WGCNA、GSEA及Western Blot实验证明NORAD通过PI3K/AKT信号通路促进LUAD的发生发展。结论:NORAD的过表达可能导致LUAD患者的不良预后。NORAD可通过PI3K-AKT-mTOR信号通路影响LUAD细胞株的基因组稳定性,从而促进LUAD的发生发展和转移。
Objective:To investigate the role of long non-coding RNA(lncRNA)NORAD in lung adeno-carcinoma(LUAD)development and its potential mechanism.Methods:By analyzing the expres-sion of NORAD in 19 cases of lung adenocarcinoma and adjacent normal tissues,we demonstrated the relationship between NORAD expression and clinicopathologic stage.The expression of NORAD in A549 and NCI-H1299 cells was detected by RT-PCR.si-NORAD interference silenced A549 and NCI-H1299 cells,and the effects of si-NORAD on cell proliferation,migration and invasion were de-tected.Western Blot was used to detect the expression of MMP9,MMP2,Vimentin,N-Cadherin,and E-Cadherin;based on the TCGA database,LUAD transcriptome data were extracted for Weight-ed Correlation Network Analysis(WGCNA).Further Gene Set Enrichment Analysis(GSEA)was used to analyze the possible signaling pathways of NORAD promoting LUAD,and then Western Blot was used to validate the effect.Results:NORAD was highly expressed in lung adenocarcinoma and prone to lymph node metastasis.NORAD was significantly overexpressed in A549 and NCI-H1299 cells,and the knockdown of NORAD significantly attenuated the proliferation,migration,and invasion of A549 and NCI-H1299 cells.Knockdown of NORAD up-regulated the expression of MMP9,MMP2,Vimentin,and N-Cadherin,and down-regulated the expression of E-Cadherin.The results of WGCNA,GSEA,and Western Blot experiments showed that NORAD promoted the development of LUAD through the PI3K/AKT signaling pathway.Conclusion:Overexpression of NORAD in LUAD could lead to a poor prognosis for LUAD patients.NORAD could affect the ge-nome stability of LUAD cell lines through PI3K-AKT-mTOR transduction,thereby promoting the occurrence and metastasis of LUAD.
作者
冉勇
石红杰
施嘉俊
董哲
赵金平
RAN Yong;SHI Hongjie;SHI Jiajun;DONG Zhe;ZHAO Jinping(Dept.of Thoracic Surgery,Zhongnan Hospital of Wuhan University,Wuhan 430071,Hubei,China)
出处
《武汉大学学报（医学版）》
CAS
2024年第4期435-441,共7页
Medical Journal of Wuhan University
