The Role And Mechanism Of ZNF831 During The Process Of Chemical-induced Lung Adenocarcinoma

Objective:1.To analyze the expression of ZNF831 gene in clinical lung adenocarcinoma patients and its relationship with clinicopathological parameters and survival time.2.To investigate the expression changes of ZNF831 gene during the process of lung adenocarcinoma induced by chemical carcinogens and its expression in different lung adenocarcinoma cell lines.3.To preliminarily explore the main functions and molecular mechanisms of ZNF831 gene in the occurrence and development of lung adenocarcinoma.Methods:1.The expression of ZNF831 in tumor tissues and adjacent normal tissues of lung adenocarcinoma patients and its relationship with clinicopathological parameters and postoperative survival and prognosis were analyzed based on the TCGA database.K-M plotter database was used to verify the data of survival and prognosis.2.QRT-PCR and RT-PCR were used to detect the expression of ZNF831 gene in HBE malignant transformation cells induced by chemical carcinogen 3-MCA and in different lung adenocarcinoma cell lines.Quantitative RT-PCR was used to analyze the expression of ZNF831 gene in lung cancer cell lines after treatment with demethylation agent 5-Aza-dC.3.Overexpression and knockdown expression models of ZNF831 gene in different lung cancer cells were established and used to analyze the role of ZNF831 gene during the development of lung adenocarcinoma.Cell proliferation and growth were detected by CCK-8 and plate colony formation experiments.Cell cycle and apoptosis were detected by flow cytometry.Cell migration and invasion were analyzed by wound healing experiments and Transwell experiments.4.The effect of ZNF831 gene on tumor growth,invasion and migration in vivo was analyzed by subcutaneous tumor formation and tail vein injection tumor formation experiments in nude mice.5.QRT-PCR and Western Blot were used to detect the mRNA and protein expression levels of downstream genes regulated by ZNF831 gene in lung adenocarcinoma,and to identify the main signaling pathways.Results:1.In the TCGA database,the expression level of ZNF831 in tumor tissues of lung adenocarcinoma patients was significantly lower than that in adjacent normal tissues(P < 0.05).The expression of ZNF831 gene was highly correlated with T stage,N stage and tumor stage(P < 0.05).Kaplan Meier analysis showed that the survival time of patients with high expression of ZNF831 gene was significantly higher than that of patients with low expression of ZNF831 gene(P < 0.05).Single factor and multivariate Cox regression analysis showed that ZNF831 expression was an independent protective factor in patients with lung adenocarcinoma(P < 0.05).2.With the increase of HBE malignant transformation algebra induced by 3-MCA,the expression level of ZNF831 gene was down-regulated gradually(P < 0.01).3.The expression of ZNF831 gene was different in different lung adenocarcinoma cell lines.Compared with HBE cell lines,ZNF831 gene was only highly expressed in A549 cell lines and low in SPC-A-1,LTEP-a-2,NCI-H1650 and other lung adenocarcinoma cell lines(P < 0.05).The expression of ZNF831 gene was up-regulated after demethylation agent 5-Aza-dC treatment.4.The overexpression of ZNF831 gene promoted the apoptosis of SPC-A-1 and LTEP-a-2 cells,inhibited cell proliferation,clone formation,invasion and migration,and blocked cells in G1 phase(P < 0.01).5.After interfering with the expression of ZNF831 in A549 cells,the ability of cell proliferation,clone formation,invasion and migration were significantly enhanced(P < 0.01).6.Interference with ZNF831 gene significantly promoted tumor growth,invasion and migration in nude mice.7.The anti-tumor effect of ZNF831 gene is mainly related to Wnt/?-catenin signaling pathway in lung adenocarcinoma.ZNF831 gene inhibited Wnt/?-catenin signaling pathway,down-regulate the mRNA and protein expression of CTNNB1(?-catenin)and its downstream target molecules MMP2,MMP7 and C-MYC.Knockdown of ZNF831 gene expression promoted the mRNA and protein expression of CTNNB1,MMP2,MMP7 and C-MYC.Treatment with Wnt/?-catenin signal pathway inhibitor significantly diminished the effect of ZNF831 in lung adenocarcinoma.Conclusions:1.ZNF831 gene was downregulated in lung adenocarcinoma.Low expression of ZNF831 gene was positively correlated with the survival and prognosis of lung adenocarcinoma patients.2.The expression of ZNF831 gene was down-regulated in 3-MCA-induced HBE cells with the increase of malignant transformation algebra.ZNF831 gene expression downexpressed in lung cancer cell lines associated with DNA methylation.3.ZNF831 gene inhibited the proliferation,invasion and migration of lung adenocarcinoma cells,and blocked the G1 phase of cells.4.The inhibitory effect of ZNF831 gene on lung adenocarcinoma is mainly related to the regulation of Wnt/?-catenin signaling pathway and downstream key molecules.