Necroptosis-related genes are associated with prognostic features of kidney renal clear cell carcinoma

Abstract

Abstract Introduction: Renal clear cell carcinoma is a kind of typical adult urological system cancer, with 30% of patients developing metastasis and 60% dying 1–2 years after diagnosis. Currently, the progress of tumor immunology and necroptosis has brought new ideas for the kidney cancer therapy. Hence, it is very necessary to find potential targets for immunotherapy combined with necroptosis. Materials and methods Based on GSE168845 and necroptosis-related genes, necroptosis-related differentially genes were identified. The prognostic value of DEGs was detected through differential expression analysis, prognostic analysis, as well as Cox regression analysis. The GSE53757 dataset was applied to verify MYCN and CDKN2A expression level. In our cohorts, the association among DEGs and clinicopathological features and overall survival was analyzed. Subsequently, the lasso Cox regression model was constructed to assess the correlation of DEGs with immune score, ICP and OCLR score. To detect the expression of two genes (MYCN, CDKN2A and ZBP1) in KIRC, the qRT-PCR were conducted in different samples. We verified the expression levels of two genes in a normal kidney cell line (HK-2 cells) and two KIRC cell lines (786-O, ACHN). Then, the protein level of MYCN and CDKN2A were detected by IHC. SiRNA was used to silence MYCN and CDKN2A expression in the ACHN cell line. Cell migration was measured by wound healing assays. Results The MYCN, CDKN2A and ZBP1 were necroptosis related genes with independent prognostic value, by which a risk prognostic model was determined. In the GSE53757 dataset, the expression of the CDKN2A gene was significantly higher in KIRC tissues compared to the normal tissues, and the expressions of the MYCN gene was significantly lower in KIRC tissues. The MYCN and CDKN2A expression was related to T, M stage and OS in our cohort. The MYCN, CDKN2A and ZBP1 were obviously correlated with immune score, ICP and OCLR score. The expression levels of CDKN2A and ZBP1 were obviously enhanced in KIRC cells than the normal kidney cells, while the expression of MYCN was decreased in KIRC cells. MYCN and CDKN2A were detected with the same results in two kinds of tissues, while ZBP1 was not obviously different. Then, the protein expression of MYCN and CDKN2A were detected by IHC,and found that MYCN and CDKN2A were obviously enhanced in KIRC tissues. The results of wound healing assay showed that, the cell migration ability of the si-CDKN2A group was significantly inhibited and the cell migration property of si-MYCN group was obviously enhanced. Conclusions MYCN and CDKN2A are potential targets and valuable prognostic biomarkers for immunotherapy combined with necroptosis in kidney renal clear cell carcinoma. CDKN2A promotes the migration of renal cancer cells. MYCN INHIBITs the migration of renal cancer cells.