转录因子ZEB1对肺腺癌细胞增殖、迁移和侵袭作用机制的研究OA

Objective To investigate the effects of zinc finger E-box binding homeobox 1(ZEB1)on the prolifera-tion,migration,and invasion of lung adenocarcinoma H322 cells,as well as its underlying molecular mechanisms.Methods The gene expression characteristics of the transcription factor ZEB1 in lung adenocarcinoma were ana-lyzed using data from the GEO and TCGA public databases.RT-qPCR and Western blot were employed to measure mRNA and protein expression levels of ZEB1 in lung adenocarcinoma cell lines(H322,A549,95-D)and normal human bronchial epithelial cells(BEAS-2B).Lentiviral transduction was utilized to establish stable ZEB1-overex-pressing(Oe-ZEB1)and vector control(Oe-NC)H322 cell lines.Cell proliferation was assessed using CCK-8,colony formation,and EdU assays,while apoptosis was evaluated by Hoechst33258/PI double staining.Wound healing and Transwell assays were performed to examine cell migration and invasion capabilities.Cell cycle distri-bution was determined by flow cytometry,and Western blot was used to analyze protein expression changes in rel-evant signaling pathways.Results The findings from GEO and TCGA indicated that ZEB1 expression in lung ad-enocarcinoma varied with tumor malignancy grade.RT-qPCR and Western blot analyses revealed significantly higher ZEB1 expression in lung adenocarcinoma cell lines compared to BEAS-2B cells(P<0.05).Results from the CCK-8,colony formation,EdU,wound healing,and Transwell assays demonstrated that,compared with the un-transfected control(Control)group,Oe-ZEB1 H322 cells exhibited enhanced proliferation,migration,and in-vasion capabilities(P<0.05).Hoechst33258/PI double staining and flow cytometry analyses showed that,rela-tive to the Control group,apoptosis was reduced in Oe-ZEB1 H322 cells(P<0.05).Additionally,a decreased proportion of cells in the G1 phase and an increased proportion in the S phase were observed in Oe-ZEB1 cells,indi-cating accelerated cell cycle progression.Western blot analysis further revealed that,compared with the Control group,Oe-ZEB1 H322 cells exhibited upregulated expression of N-cadherin,mutant p53(mutp53),and Cyclin D1(P<0.05),while expression levels of E-cadherin,murine double minute 2(MDM2),and p21 were down-regulated(P<0.05).Conclusion Overexpression of ZEB1 promotes the proliferation,migration,and invasion of lung adenocarcinoma H322 cells and may facilitate cell cycle progression by modulating the MDM2/mutp53/p21 signaling pathway,thereby promoting the transition of cells from the G0/G1 phase to the S phase.