Screening Of Lung Adenocarcinoma-Associated Gene TRIM9 Based On High-Throughput Sequencing And Its Correlation With Clinical Parameters

Objective:To investigate the expression of TRIM9 gene in lung adenocarcinoma and its correlation with clinicopathologic factors.Methods:Four patients with lung adenocarcinoma who were diagnosed between January 2020 and December 2020 in the Third Clinical Medical College of Xinjiang Medical University and four healthy subjects who visited our hospital for physical examination at the same time were selected.(1)Illumina high-throughput sequencing(RNA-Seq)was used to extract total m RNA and quality inspection of the collected samples,and the data after quality control were compared with the reference genome sequence,Pearson correlation test was used to assess sample consistency,| log2 FC | ≥ 1,P ≤ 0.05 was used as the screening criteria for differential gene m RNA expression profiling,and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases were used to analyze differential gene-related enrichment and signaling pathways.(2)TRIM9,which is most closely related to lung adenocarcinoma,was selected,and the expression of TRIM9 in different lung adenocarcinoma cell lines and human pulmonary artery smooth muscle cells was verified by q RT-PCR,while 15 cases of lung adenocarcinoma and 15 cases of peripheral blood of healthy subjects were collected to verify the expression level of TRIM9 and analyze the correlation between TRIM9 expression and clinical factors.Results:(1)A total of 623 differentially expressed genes(DEGs)were differentially expressed in lung adenocarcinoma compared with healthy subjects,of which 318 were up-regulated differential genes and 305 were down-regulated differential genes;significantly upregulated genes were OLAH,AC090587.2,TRIM9,etc.;and significantly down-regulated genes were AC090772.1,DSC1,BHLHA15,etc.The GO database results showed that DEGs were expressed in many different functional domains in lung adenocarcinoma,and the top five functional enrichment were mainly concentrated in mitochondrial ATP synthesis coupled with electron transport,nucleosomes,respiratory chain NADH dehydrogenase activity and other functions in biological pathways.KEGG analysis showed that DEGs were mainly involved in various pathways such as systemic lupus erythematosus,oxidative phosphorylation,and Parkinson’s disease.(2)The results of cell level showed that the expression of TRIM9 was different in lung adenocarcinoma cell lines compared with human pulmonary artery smooth muscle cells,and the expression level was the highest in A549 cells;the results of tissue level showed that the relative expression level of TRIM9 was increased in the lung adenocarcinoma group compared with the healthy control group(P ≤ 0.05),and in the lung adenocarcinoma group,the expression level of TRIM9 was negatively correlated with the carcinoembryonic antigen(CEA)level(P > 0.05),but not significantly correlated with the age,gender,smoking index,and tumor stage of the patients(P > 0.05).Conclusion:(1)Differential gene expression profiles were selected from patients with lung adenocarcinoma,of which OLAH,AC090587.2,TRIM9,AC090772.1,DSC1 and other genes may be involved in the occurrence and development of lung adenocarcinoma;(2)GO and KEGG analysis showed that differential genes may be involved in nucleosomes,oxidative phosphorylation and other processes,involving systemic lupus erythematosus,oxidative phosphorylation and other related pathways;(3)TRIM9 expression levels were the highest in A549 cells,and the expression levels in the peripheral blood of patients with lung adenocarcinoma were significantly higher than those in healthy controls,while its high expression was negatively correlated with CEA in the peripheral serum tumor marker indicators of patients with lung adenocarcinoma,so we believe that TRIM9 may be involved in the occurrence and development of lung adenocarcinoma.