Identification And Mechanism Of Key Genes In The Efficacy Of Radiotherapy For Cervical Cancer

Objective:To explore the key genes related to the efficacy of radiotherapy for cervical cancer.To explore the potential mechanisms affecting the efficacy of radiotherapy for cervical cancer.Methods:Tissue samples were collected from 20 cervical cancer patients undergoing radiotherapy.Tumor tissue samples were taken before and after radiotherapy to evaluate the therapeutic effect and perform tissue sequencing.The samples of cervical cancer treated with radiotherapy in The Cancer Genome Atlas were downloaded and collated.The sequencing data of cervical normal tissues,cervical intraepithelial neoplasia and cervical cancer tissues from Gene Expression Omnibus data were downloaded.Various immune signatures and marker genes in the tumor immune microenvironment were obtained by searching the literature.Single Sample Gene Set Enrichment Analysis was used to calculate the enrichment score of immune signatures in all samples.Random Forest was used to construct an efficacy classifier of immune signatures to screen the immune signatures that contributed greatly to the classification of radiotherapy efficacy in cervical cancer.Then,the signatures that changed significantly before and after radiotherapy were screened as the key signatures affecting the efficacy of radiotherapy.Weighted Gene Co-expression Network Analysis was used to identify the hub genes that were significantly positively correlated with the key signatures.Enrichment analysis was performed on the differentially expressed genes before and after radiotherapy to obtain the pathways activated by radiotherapy.The activation degree was quantified by Single Sample Gene Set Enrichment Analysis to screen the pathways with significant differences in different therapeutic groups.According to the expression matrix of each sample,the immune score of cervical cancer tissue and the proportion of immune cell infiltration in the immune microenvironment were calculated.Finally,the correlation between hub genes and pathways,hub genes and immune cell infiltration was analyzed to explore the potential mechanisms affecting the efficacy of cervical cancer radiotherapy.Results:FcR-Like Protein 5(FCRL5),Tumor Necrosis Factor Receptor Superfamily Member 17(TNFRSF17),Pim-2 Proto-Oncogene were identified by bioinformatics analysis.Serine/Threonine Kinase,(PIM2),Sterile Alpha Motif Domain-Containing Protein3(SAMD3).These genes are hub genes affecting the efficacy of radiotherapy.There were significant differences in the expression of four hub genes in the samples before and after radiotherapy and in different efficacy groups.These genes were highly expressed after radiotherapy and in the samples from the better response group.Among them,TNFRSF17 and PIM2 also showed an increased expression trend in the cervical cancer development group(normal cervical tissue,cervical intraepithelial neoplasia tissue,cervical cancer tissue).The pathways related to radiotherapy efficacy were identified by enrichment analysis,and then the activation of pathways was quantified to explore their correlation with hub gene expression.Four pathways were identified to be significantly associated with hub genes.The activation levels of Cytokine-cytokine receptor interaction,Chemokine signaling pathway,Cell adhesion molecules and B cell receptor signaling pathway increased with the expression of the four hub genes.The results of immune infiltration and immune score analysis showed that CD4+T cells,CD8+T cells,plasma cells,naive B cells and macrophages increased with the expression of hub genes,while dendritic cells decreased with the expression of Hub gene.Anti-tumor T cells and plasma cells responsible for humoral immunity were elevated.M1 macrophages,the classic immune-promoting cells,also showed an elevated status.As antigen presenting cells,dendritic cells showed a decreased state.The decrease of naive B cells and M0 cells with the increase of hub gene expression also indirectly reflects the change of immune microenvironment.Conclusions:The expression of FCRL5,TNFRSF17,PIM2 and SAMD3 has an important impact on the efficacy of radiotherapy for cervical cancer.FCRL5,TNFRSF17,PIM2 and SAMD3 are significantly up-regulated during radiotherapy,and their high expression indicates a better radiotherapy efficacy.The infiltration of some anti-tumor immune cells and the activation of immune-related pathways were increased with the increase of the expression levels of FCRL5,TNFRSF17,PIM2 and SAMD3.In the course of radiotherapy for cervical cancer,the immune response is enhanced and suppressed,but in general,it is enhanced.