Construction of CXCR2 gene knockout HeLa cell line and transcriptome sequencing analysis
Objective:To investigate CXCR2 related functional genes and signaling pathways through gene knockout and transcriptome sequencing,revealing its molecular role in cervical cancer.Methods:The CRISP R-Cas9 knockout plasmid vector targeting CXCR2 gene was designed and constructed.HeLa cells were transfected and CXCR2 gene deletion cell lines were screened.Subsequently,the differentially expressed genes were analyzed by transcriptome sequencing,and the signaling pathways and important differentially expressed genes were screened by gene ontology(GO)and kyoto encyclopedia of genes and genome(KEGG)enrichment analysis.The genes were verified by real-time fluorescence quantitative PCR.Results:The CXCR2 gene knockout cells were successfully constructed,and 1 519 differentially expressed genes were obtained by transcriptomic sequencing,of which 428 were up-regulated and 1 091 were down-regulated.Analysis of KEGG signaling pathway enrichment showed that the tumor pathway was the most enriched signaling pathway,followed by the PI3K-Akt signaling pathway and HPV infection pathway.Seven common differentially expressed genes including EGF,FN1,ITGA2B,CCND3,PIK3CD,PDGFRB and CDKN1A were obtained.Among them,the expression of the first six genes was significantly down-regulated,while the expression of CDKN1A was significantly up-regulated.Conclusions:The results of transcriptome analysis further prompts the role of CXCR2 in cervical cancer,and preliminarily identifies its related signaling pathways and key functional genes,providing a theoretical basis for the molecular mechanism and clinical research of cervical cancer.
国家科技期刊平台
NSTL
万方数据
维普
