摘要
目的长链非编码RNA(lncRNA)SATB2的反义转录物(SATB2-AS1)对肺腺癌细胞生物学功能的影响和机制研究。方法收集25例肺腺癌患者的肿瘤组织和癌旁组织,检测SATB2-AS1表达水平。用lncRNA SATB2-AS1的过表达载体(pcDNA-SATB2-AS1)和shRNA或IGF2BP2的shRNA(sh-SATB2-AS1;sh-IGF2BP2)和/或SLC7A11的shRNA(sh-SLC7A11)转染肺腺癌细胞A549。RIP分析评估IGF2BP2蛋白分别与SATB2-AS1或SLC7A11 mRNA的结合;CCK-8和Transwell实验分别用于检测肺腺癌细胞的增殖和侵袭能力;RT-qPCR和Western blot分别检测基因和蛋白表达。结果与癌旁组织和人正常支气管上皮细胞系BEAS-2B相比,SATB2-AS1在肺腺癌患者的肿瘤组织和肺腺癌细胞系中的表达显著下调(P<0.01)。过表达SATB2-AS1抑制A549细胞增殖和侵袭,沉默SATB2-AS1促进细胞增殖和侵袭(P<0.05)。过表达SATB2-AS1后A549细胞中的Fe2+浓度、活性氧(ROS)水平以及丙二醛(MDA)含量显著降低(P<0.01),还原性谷胱甘肽(GSH)含量及铁死亡关键蛋白SLC7A11、GPX4的表达显著升高(P<0.01)。SATB2-AS1通过与IGF2BP2蛋白结合显著促进IGF2BP2蛋白与SLC7A11 mRNA结合,降低SLC7A11 mRNA的稳定性(P<0.01)。沉默SLC7A11显著逆转了SATB2-AS1沉默对A549细胞的影响。结论lncRNA SATB2-AS1通过招募IGF2BP2蛋白破坏SLC7A11mRNA稳定性,诱导肺腺癌细胞铁死亡,抑制细胞增殖和侵袭,从而抑制肺腺癌进展。
Objective To investigate the effect of long non-coding RNA(lncRNA)antisense transcript of SATB2 protein(SATB2-AS1)on the biological function of lung adenocarcinoma cells and its mechanism.Methods Tumor tissues and adjacent tissues from 25 patients with lung adenocarcinoma were collected to detect the expression of SATB2-AS1.Then,the overexpression vector of lncRNA SATB2-AS1(pcDNA-SATB2-AS1)and shRNA or shRNA of IGF2BP2(sh-SATB2-AS1;sh-IGF2BP2)and/or shRNA of SLC7A11(sh-SLC7A11)were transfected into lung adenocarcinoma cell A549.RIP assay was used to estimate the binding of IGF2BP2 protein to SATB2-AS1 or SLC7A11 mRNA,respectively;CCK-8 and Transwell assay were used to detect the proliferation and invasion ability of lung adenocarcinoma cells;RT-qPCR and Western blot assay were used to detect gene expression and protein expression,respectively.Results Compared with adjacent tissues and human bronchial epithelial cell BEAS-2B,the expression of SATB2-AS1 was significantly down-regulated in tumor tissues of patients with lung adenocarcinoma and lung adenocarcinoma cells(P<0.01).Overexpression of SATB2-AS1 significantly inhibited the proliferation and invasion of A549 cells,while silencing SATB2-AS1 significantly promoted the cell proliferation and invasion(P<0.05).Moreover,overexpression of SATB2-AS1 significantly reduced Fe2+concentration,reactive oxygen species(ROS)level,and malondialdehyde(MDA)content in A549 cells(P<0.01),and increased glutathione(GSH)content,the expression of SLC7A11 and GPX4 which were the key proteins of ferroptosis(P<0.01).Meanwhile,SATB2-AS1 significantly promoted IGF2BP2 protein binding to SLC7A11 mRNA by binding to IGF2BP2 protein,and reduced the stability of SLC7A11 mRNA(P<0.01).Silencing SLC7A11 significantly reversed the effects of silencing SATB2-AS1 on A549 cells.Conclusion LncRNA SATB2-AS1 destabilizes SLC7A11 mRNA by recruiting IGF2BP2 protein,induces the ferroptosis of lung adenocarcinoma cells,inhibits the cell proliferation and invasion,and thus inhibits the progression of lung a
作者
乔勃伟
张勇
李明涛
李岩
QIAO Bo-wei;ZHANG Yong;LI Ming-tao;LI Yan(Department of Thoracic Surgery,the Second Affiliated Hospital of Air Force Medical University,Xi'an Shaanxi 710038,China)
出处
《局解手术学杂志》
2024年第10期876-882,共7页
Journal of Regional Anatomy and Operative Surgery
基金
陕西省自然科学基础研究计划项目(2020JQ-920)。
