| Objective:According to recent research,the tumor microenvironment(TME)is crucial to the development of tumors.However,more research is required to determine the function of the TME in renal cell carcinoma(RCC)and how to use processes associated to the TME to inhibit the proliferation and invasion of renal cell carcinoma cells.It has been reported that CD3 E is closely related to the immune microenvironment of muscle-invasive bladder cancer,and is expected to be a prognostic indicator for muscle-invasive bladder cancer.It is yet unknown how CD3 E and the immune microenvironment of renal cell cancer relate to one another.This investigation’s goals are to learn more about the relationship between CD3 E and the immunological milieu of RCC and the possible applications of CD3 E in the Prognostic assessment and treatment of RCC.Methods:In this study,we obtained transcriptome and clinical information from the Cancer Genome Atlas(TCGA)database for 893 patients with RCC.The downloaded data was then combined and integrated using Perl,and the immunological and matrix components of the samples’ microenvironments were scored using R and the ESTIMATE calculating method.The samples were then separated into two groups based on the median score.According to the difference in clinical information and survival between the two groups,the score was analyzed in terms of prognosis and clinical correlation with renal cell carcinoma patients.After that,the differences in gene expression were analyzed for the high and low scores of immunity and matrix scores,and the gene sets obtained were intersected to obtain differentially expressed genes(DEGs).Next,we perform GO and KEGG enrichment analysis on DEGs to obtain gene set-related functions.After that,we used the string database to construct a protein-protein interaction(PPI)network related to DEGs to obtain the core genes of the network,performed univariate COX regression analysis on kidney cancer samples to obtain prognostic related genes,and intersected the above two sets of genes to obtain predictors-CD3 E and LCK.After that,we obtained the difference between LCK and CD3 E expression in kidney cancer and normal renal samples through differential analysis,and obtained their correlation with the survival of kidney cancer patients.We used the real-time quantitative polymerase chain reaction(q-PCR)experimental method to verify the differences in the expression levels of CD3E and LCK we obtained,and finally determined that CD3E was a predictive gene related to the prognosis of RCC according to the correlation of the test.Next,we used Western Blot(WB)and immunohistochemistry to further verify the high expression of CD3E in renal cancer cells.We divided the samples into two groups with high and low expression according to the expression of CD3E,and analyzed the clinical information of the samples to obtain the correlation between CD3E and the clinical information of patients with RCC.Based on the expression level of CD3E,we performed gene set enrichment analysis(GSEA)on the samples.We used CIBERSORT to calculate the proportion of tumor-infiltrating immune cells(TIC)in kidney cancer samples,and analyzed the correlation and difference between the proportion of TICs and the expression of CD3E,and finally obtained the correlation between CD3E and immune cells in the immune microenvironment.Results:The scores obtained based on the immune and stromal components of RCC samples had a significant negative correlation with the survival of kidney cancer patients,and the scores obtained based on immune components were more significant.Through differential analysis,we obtained a total of 1755 DEGs,and GO and KEGG enrichment analysis of DEGs showed that their overall function was mainly related to the immune activity of renal cell carcinoma.We further analyzed the selected CD3 E and LCK genes,and compared with normal kidney tissue,CD3 E and LCK were significantly higher expressed in RCC tissue,and the expression of genes in kidney cancer tissue was significantly upregulated compared to adjacent tissues in the same sample.The survival analysis of single genes showed that the expression of CD3 E and LCK had a significant negative correlation with the survival of kidney cancer patients,and the higher the expression of genes,the worse the prognosis of patients.We used q-PCR experiments to verify that CD3 E and LCK in renal cell cancer cells are highly expressed at the cellular level,and we found that the difference in CD3 E was more pronounced.We selected CD3 E for further WB and immunohistochemistry experiments,thereby verifying the high expressibility of CD3 E in kidney cancer patients at the tissue level.The correlation analysis of CD3 E and clinical information of renal cell carcinoma showed that there was a significant positive correlation between CD3 E and the stage and grade of renal cell carcinoma patients.Gene enrichment analysis was carried out on the high and low expression groups based on CD3 E expression group,and it can be seen that CD3 E is mainly manifested as immune-related enrichment.Through the correlation and differential analysis of immune cell infiltration and CD3 E in renal cell carcinoma,14 immune cells in the immune microenvironment had a significant correlation with CD3 E,8 kinds of immune cells were positively correlated,and 6 immune cells had obvious negative correlation with CD3 E.Conclusions:CD3E is correlated with the immune microenvironment of renal cell carcinoma,is a prognostic biomarker for patients with RCC,and is expected to participate in the treatment of RCC. |
PDF Full Text Request