Target Gene Analysis Of ESRP1 In Cutaneous Melanoma And Its Effect On Tumor Biological Behaviors

Objective:By using modern data mining technology,bioinformatics analysis was performed on the expression and function regulation of the epithelial splicing regulator proteinl(ESRP1)in cutaneous melanoma,and ESRP1 related kinase,miRNA and transcription factor protein interaction network was constructed.Methods:From the transcriptional sequencing data of 103 cutaneous melanoma samples in the Cancer Genome Atlas database,the expression level of ESRP1 and its gene regulation networks were analyzed.LinkedOmics was used to identify differential gene expression with ESRP1 and to analyze Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways.Gene enrichment analysis examined target networks of kinase,miRNA and transcription factor.Results:The expression of ESRP1 in cutaneous melanoma was lower than that in normal tissues,and its expression was inversely proportional to the malignant degree such as tumor stage and lymph node metastasis.The first 50 important genes positively and negatively related to ESRP1 were extracted.Functional network analysis suggested that ESRP1 regulated epidermal development,translation initiation,ribosome,drug metabolism and chemical carcinogenesis via pathways involving several cancer-related kinases,miRNAs and transcription factors.The kinase-target networks significantly associated with ESRP1 were CDK1,GRK3,PRKACB,PRKACK,and PRKX.miRNA-target networks included MIR-138,MIR-205,MIR-219,MIR-453 and MIR-423.Transcription factor-target networks included the ETF,E2F,EN,USF,and CEBPB transcription factor families.Conclusion:Our results demonstrated that data mining efficiently revealed information about ESRP1 expression and potential regulatory networks in cutaneous melanoma,laying a foundation for further study of the role of ESRP1 in carcinogenesis.Chapter 2 Effects of ESRP1 on the biological behaviors of cutaneous melanoma cellsObjective:ESRP1 was described as an epithelial-specific splicing regulator involved in tumorigenesis and development,but its role in cutaneous melanoma was unclear.This section aimed to explore the effect of ESRP1 on the biological behaviors of cutaneous melanoma cells and its potential regulatory mechanism.Methods:We detected the expression level of ESRP1 in skin melanoma tissues and cells by real-time quantitative PCR and western blot,and used lentiviral transfection technology to overexpress and interfere with ESRP1 gene.CCK8 method was used to detect cell proliferation ability in vitro,the cell invasion and metastasis ability were detected by transwell,and western blot was used to detect the expression of epithelial and mesenchymal transformation phenotypic proteins.Results:The expressions of ESRP1 in metastatic cutaneous melanoma tissues were lower than that in primary tumors,which was associated with epithelial-mesenchymal transformation phenotypic proteins.We found that the proliferation,invasion and metastatic ability of ESRP1-shRN A melanoma cells was significantly enhanced compared with the blank control and negative control groups(p<0.05).Meantime,the expression of E-cadherin was reduced,N-cadherin and Vimentin were increased,while the results obtained by overexpressing ESRP1 were the opposite.Conclusion:ESRP1 was lowly expressed in metastatic cutaneous melanoma tissues,which is consistent with bioinformatics analysis.The ability of melanoma cells with low expression of ESRP1 to proliferate,invade and metastasize was significantly enhanced.This process may be regulated by epithelial-mesenchymal transformation.