Comprehensive Analysis And Targeted Drug Screening Of Oncogene AURKA In Cutaneous Melanoma Based On Bioinformatics And Molecular Docking Technology

Background:Cutaneous melanoma(CM)is a highly invasive and drug-resistant form of skin tumor with an insidious onset.It has the highest fatality rate among skin tumors.While surgical resection can cure early primary cutaneous melanoma,it is prone to metastasis,and the 5-year survival rate drops to only 15~20%when accompanied by lymph node or distant metastasis.In recent years,significant progress has been made in the treatment of advanced melanoma through the use of immunotherapy and targeted therapy.However,it is important to note that only a minority of CM patients(around 30%to 40%)are able to benefit from these treatments.Furthermore,more than half of CM patients experience high levels of drug resistance and serious side effects when undergoing immune checkpoint blockade therapy(ICB)or targeted therapy.As a result,the clinical application of these treatments has been limited.This study aims to investigate potential biomarkers and therapeutic targets that can predict the prognosis of patients with CM.Specifically,we seek to understand the expression pattern,prognostic value,immune response,biological function,and potential targeted drugs of the Aurora kinase(AURKA).Methods:1 Identification of the core hub genes of CM:1.1 To identify hub genes specific to cutaneous melanoma(CM),we analyzed original data sets of both CM and normal skin tissues.We utilized various techniques such as differential expression analysis,weighted gene coexpression network analysis(WGCNA),protein interaction analysis,and four screening HUB gene topology algorithms.Through the use of bioinformatics methods,we were able to successfully screen and identify CM hub genes.1.2 In order to identify the core hub gene that affect the occurrence,development,and prognosis of CM,various analyses were conducted on the selected hub genes.These included correlation analysis,single factor COX regression analysis,and ROC diagnostic analysis.2 This subject paper aims to comprehensively analyze the impact of the core hub gene on the prognosis and immune infiltration of CM:This subject paper explores the prognostic value,potential role,and impact of the CM core hub gene through differential analysis,prognostic analysis,and functional enrichment analysis.Additionally,various immune infiltration algorithms such as CIBERSORT,TIMER,EPIC,x Cell,MCPCOUNTER,QUANTISEQ,CIBERSORT＿ABS,ss GSEA,and ESTIMATE were utilized to study the influence of the tumor microenvironment on the core hub gene.3 This study aims to discuss the screening of small molecular compounds that target the protein encoded by the core hub gene and its mechanism of action.:3.1 To identify potential inhibitors of the protein encoded by the core hub gene,we used computer molecular docking technology to screen the ZINC database for small molecule compounds.We narrowed down our search to the top 100 compounds and then conducted MTT experiments to score their effectiveness.From this,we selected the top 10compounds and subjected them to cytotoxicity assays.3.2 In this study,we intervened A875 cells with ZINC000097018978,a small molecule compound that targets the protein encoded by the core hub gene.To assess the effects of the intervention,we used flow cytometry to detect changes in the cell cycle and cell apoptosis.Additionally,we employed scratch tests to measure cell migration and Transwell tests to evaluate cell invasion ability.we utilized RT-q PCR and western blot tests to detect cells and investigate the m RNA and protein expression levels of the core hub gene,Caspase-3,Bax,Bcl-2,and CDK1.Our aim was to explore the possible mechanism of the small molecule compound ZINC000097018978 targeting the protein encoded by the core hub gene to inhibit CM.Results:1 The study identified seven HUB genes(AURKA,KIF20A,TPX2,DLGAP5,ASPM,KIF23,CDKN3)associated with CM through differential expression analysis,WGCNA,protein interaction analysis,and four topological algorithms.Further analysis through correlation analysis,univariate COX regression analysis,and ROC diagnostic analysis of these seven genes revealed that AURKA is the primary HUB gene affecting differential expression and prognosis of CM.2 AURKA is considered an independent marker of poor prognosis in patients with CM,and the use of a constructed nomogram has been shown to have better predictive performance.Through the analysis of various immune infiltration algorithms,it has been discovered that the expression of AURKA is negatively correlated with the degree of infiltration of most immune activation cells(CD8~+T,CD4~+T,B cells).Conversely,it is positively correlated with the degree of infiltration of immunosuppressive cells such as Th2and Myeloid-Derived Suppressor Cell.Additionally,AURKA is negatively correlated with anti-tumor immune cell marker genes(CD8~+T,CD4~+T,B cells),antigen presentation and processing characteristic genes,immune checkpoints,and MHC molecules.High levels of AURKA expression are observed in patients with low expression of these markers.Furthermore,single-cell analysis revealed that AURKA is predominantly expressed in CM cells,exhausted CD8~+T cells,and other cells that facilitate tumor immune evasion and worsen tumor malignancy.AURKA was observed to impact the expression of genes characteristic of hypoxia in CM,and patients with elevated AURKA expression exhibited an increased tumor stemness index.Mechanistically,AURKA overexpression was linked to significant upregulation of the TGF-βsignaling pathway,which in turn influenced the patterns of chemokines and cytokines.3 Using molecular pair technology,we screened the ZINC database for the top 10small molecular compounds that can target and inhibit AURKA expression.The results of MTT experiments revealed that these compounds had varying degrees of toxicity on A875cells.Among them,ZINC000097018978 had the strongest cytotoxic effect on A875 cells.4 ZINC000097018978 has been found to have inhibitory effects on the proliferation and invasion of A875 cells.The mechanism behind this may be attributed to the arrest of A875 cells in the G2/M phase,as well as a reduction in the anti-apoptotic protein Bcl-2and an increase in pro-apoptotic proteins that cleave Caspase-3 and Bax,leading to the induction of apoptosis in A875 cells.These findings suggest that ZINC000097018978 has the potential to inhibit tumor cell proliferation and invasion.Conclusions:1 AURKA is likely the central gene responsible for driving the occurrence and development of CM.2 The overexpression of AURKA in CM patients indicates the development of a hypoxia-induced tumor microenvironment that is immunosuppressive.This discovery makes AURKA a promising biomarker for prognosis and immune infiltration,as well as a potential therapeutic target.3 ZINC000097018978 is an effective inhibitor of AURKA,which can inhibit the proliferation,invasion of A875 cells.