摘要
目的探讨miR-9500通过靶向SMAD2调控肺腺癌细胞迁移侵袭的相关分子机制。方法通过生物信息学分析筛选出miR-9500的核心靶基因,并对其进行GO功能和KEGG信号通路富集及生存分析。预测miR-9500与其关键靶基因SMAD2之间的靶向结合位点,双荧光素酶报告实验验证miR-9500与SMAD2之间是否存在直接靶向关系,qRT-PCR和Western blot检测miR-9500对SMAD2 mRNA和蛋白表达水平的影响。划痕、Transwell实验及基质胶侵袭实验分析miR-9500对肺腺癌细胞迁移侵袭能力的影响。结果miR-9500核心靶基因主要富集于癌症通路、TGF-β信号通路和黏着斑信号通路等。排名前10的核心靶基因中,只有VAMP2、SMAD2及RXRA的表达水平与肺腺癌患者的总生存期显著相关。miR-9500可靶向结合SMAD2来下调SMAD2的表达水平。且过表达miR-9500可显著抑制肺腺癌细胞的迁移和侵袭能力,并显著降低迁移侵袭标志蛋白MMP2、MMP9的表达水平。结论miR-9500可通过靶向SMAD2抑制肺腺癌细胞的迁移侵袭,其可能作为抑癌因子在肺腺癌的发生发展中发挥重要调控作用。
Objective To explore the molecular mechanism underlying miR-9500 regulating the migration and invasion of lung adenocarcinoma cells by targeting SMAD2.Methods The core target genes of miR-9500 were screened out by bioinformatics analysis,and their GO function analysis,KEGG signaling pathway enrichment,and survival analysis were performed.The targeted binding sites between miR-9500 and SMAD2 were predicted,and the direct targeting relationship between miR-9500 and SMAD2 was verified by dual-luciferase reporter assay.qRT-PCR and Western blot were used to assess the effect of miR-9500 on the mRNA and protein expression levels of SMAD2.Wound healing assay,Transwell assay,and Matrigel invasion assay were used to determine the effect of miR-9500 on the migration and invasion of lung adenocarcinoma cells.Results The core target genes of miR-9500 were mainly enriched in the cancer pathway,TGF-βsignaling pathway,and focal adhesion.However,only the expression levels of VAMP2,SMAD2,and RXRA among the top 10 core target genes were significantly correlated with the overall survival of patients with lung adenocarcinoma.miR-9500 targeted SMAD2 and down-regulated the expression levels of SMAD2,and overexpression of miR-9500 significantly inhibited the migration and invasion of lung adenocarcinoma cells and markedly decreased the expression levels of MMP2 and MMP9.Conclusion miR-9500 can inhibit the migration and invasion of lung adenocarcinoma cells by targeting SMAD2,which may play an important role in the tumorigenesis and development of lung adenocarcinoma as a tumor suppressor.
作者
李芳
韩采利
王丽
沈慧
LI Fang;HAN Caili;WANG Li;SHEN Hui(School of Basic Medical Sciences,Medical School of Yan'an University,Yan'an 716000,China)
出处
《肿瘤防治研究》
CAS
2023年第3期236-242,共7页
Cancer Research on Prevention and Treatment
基金
陕西省自然科学基础研究计划(2022 JQ-907)
陕西省高校科协青年人才托举计划(20210309)
2021年国家级大学生创新创业训练计划(202110719015)
延安大学研究生教育创新计划项目(YCX2021088)。
