Correlations of coactivator-associated arginine methyltransferase 1 in renal clear cell carcinoma with epithelial-mesenchymal transition and immune cell infiltration
Objective To observe the expression of coactivator-associated arginine methyltransferase 1(CARM1)in renal clear cell carcinoma and to explore its impact on epithelial-mesenchymal transition(EMT)and immune cell infiltration.Methods Caki-1 cells in logarithmic growth phase were randomly divided into CARM1 knockdown group(transfected with CARM1 knockdown lentivirus),overexpression group(transfected with CARM1 overexpressed lentivirus),and negative control group(transfected with negative control lentivirus).The relative expression of CARM1 mRNA was detected by real-time fluorescence quantitative PCR in three groups 48 h after transfection.The number of invading cells was measured by Transwell assay.The relative expressions of CARM1,E-cadherin,N-cadherin and vimentin proteins were detected by Western blot 72 h after transfection.The Coexpedia database was utilized to select genes co-expressed with CARM1 in renal clear cell carcinoma tissues,and a protein interaction network was constructed using the STRING database.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis was performed on proteins co-expressed with CARM1.The quanTIseq tumor-infiltrating immune cell quantification analysis explored the Pearson correlation coefficients of CARM1 expression with immune cell infiltration scores in renal clear cell carcinoma,papillary renal cell carcinoma,mixed renal cell carcinoma,and chromophobe renal cell carcinoma,identifying the significantly correlated tumor immune cells of renal cancer.TIMER immune cell infiltration analysis was used to examine the correlation between CARM1 expression and levels of immune cell infiltration in different types of renal cancer.Results The relative expression of CARM1 mRNA 48 h after transfection was higher in overexpression group(3.59±0.27)than that in negative control group(1.00±0.01)and knockdown group(0.43±0.06)(P＜0.05),and higher in negative control group than that in knockdown group(P＜0.05).The relative expression of E-cadherin protein 72 h after transfection was lower in overexpression group(0.44±0.04)than that in negative control group(1.00±0.05)and knockdown group(2.06±0.10)(P＜0.05),and lower in negative control group than that in knockdown group(P＜0.05),while the relative expressions of CARM1,N-cadherin and vimentin proteins were significantly higher in overexpression group(2.15±0.19,4.44±0.39,2.87±0.23)than those in negative control group(1.00±0.07,1.00±0.05,1.00±0.07)and knockdown group(0.45±0.08,0.48±0.08,0.45±0.04)(P＜0.05),and higher in negative control group than those in knockdown group(P＜0.05).The number of invading cells 72 h after transfection was more in overexpression group(178±10)than that in knockdown group(44±4)and negative control group(83±4)(P＜0.05),and more in negative control group than that in knockdown group(P＜0.05).The Coexpedia database identified the top 100 genes co-expressed with CARM1 in renal clear cell carcinoma tissues.The protein interaction network highlighted interactions between CARM1 and transcription factors such as FOS,JUN and CREBBP,suggesting CARM1's involvement in the translation of downstream proteins.KEGG enrichment analysis indicated that CARM1 co-expressed genes were primarily enriched in the pathways as cell cycle,PI3K/Akt signaling pathway,RNA transport,and focal adhesion.The quanTIseq analysis revealed a positive correlation between CARM1 expression and neutrophil levels in different types of renal cancer.TIMER analysis showed a positive correlation of CARM1 expression with infiltration levels of B cells,CD8+T cells,CD4+T cells,macrophages,neutrophils,and dendritic cells in renal clear cell carcinoma.Conclusions CARM1 regulates EMT in renal clear cell carcinoma to promote invasion by downregulating E-cadherin and upregulating N-cadherin and vimentin.CARM1 may interact with a variety of proteins,influence celluar life activities,and participate in the immune response of various types of renal cancer.
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