Bioinformatic Analysis And Functional Study Of AURKA In Lung Adenocarcinoma

ObjectiveTo investigate the expression level and clinical significance of Auroral kinase A(AURKA)in lung adenocarcinoma(LUAD).And the purpose of this study is to clarify the effect of AURKA overexpression on the functional phenotype of LUAD cells and provide some insights into clinical treatment and drug development of patients with LUADMethod1?Bioinformatics analysisThe expression of AURKA mRNA in LUAD and normal lung tissues were analyzed by GSE10072 and GSE40791 gene chip data from GEO database and RNA-seq(HTSeq-FPKM)data from TCGA database.TCGA database was used to analyze the correlation between AURKA mRNA expression and pathological features and prognosis of patients with LUAD.GSEA enrichment analysis was performed to explore AURKA-related pathways.2?Immunohistochemistry staining(IHC)The expression of AURKA protein in LUAD and normal lung tissues was evaluated using Human Protein Atlas.3? Human LUAD cell lines A549 and H1299 were transfected with AURKA overexpression plasmid(OE-AURKA)or overexpression control plasmid(Ctrl).Then,q RT-PCR was used to evaluate transfection efficiency4?Cell experimentAnd then cell migration,proliferation,and invasion viability were,respectively,assessed by wound healing assay,CCK-8 assay,clone formation assay,and transwell assay,so as to evaluate the effect of overexpression of AURKA on the biological behavior of LUAD cells.Result1?The data analysis of GEO and TCGA database indicated that expression levels of AURKA mRNA and protein in LUAD were significantly higher than those in normal tissues.2?Kruskal-Wallis test showed that the expression of AURKA mRNA was different in TNM stages.Logistic regression showed that the expression of AURKA mRNA was significantly correlated with TNM stage,N stage,age,and gender.3?Kaplan-Meier survival analysis showed that the expression of AURKA mRNA was related to the overall survival of patients.Univariate and multivariate Cox analysis revealed that high TNM stage and high AURKA mRNA were significantly associated with poor prognosis and may be independent prognostic factors.4?GSEA enrichment analysis indicated that cell cycle,p53 pathway,small cell lung cancer,and glycolysis gluconeogenesis were mainly enriched in AURKA high expression phenotype.5?The results of cytological experiments indicated that overexpression of AURKA promoted migration,proliferation,and invasion of A549 and H1299 cells.ConclusionsThe AURKA mRNA and protein levels in LUAD was significantly higher than that in normal lung tissues,associated with aggressive progression and poor survival in patients.The elevated AURKA expression promoted migration,proliferation,and invasion of A549 and H1299 cells.Our findings indicated that AURKA may serve as independent prognostic factor and potential biomarker in LUAD.