Abstract
BackgroundAryl hydrocarbon receptor nuclear translocator like 2 (ARNTL2) is a member of the PAS superfamily. Previous studies explored the carcinogenic roles of transcription factor ARNTL2 in human malignancies. However, its roles in ccRCC have not been elucidated. This study sought to explore the roles of ARNTL2 in ccRCC and determine its correlations with tumor immunity.MethodsThe expression of ARNTL2 was analyzed using the GEO, TCGA and GTEx database, and verified in ccRCC tissue samples and cell lines by qRT-PCR and western blot analysis. Kaplan–Meier survival curve analysis, Cox regression analysis (including univariate and multivariate analysis) was utilized to evaluate the prognostic values of ARNTL2. Potential biological mechanisms of ARNTL2 were explored using GSEA method. Colony formation and wound healing assays were conducted to explore the oncogenic role of ARNTL2 in ccRCC. ssGSEA and xCell algorithm were used to explore the correlation between ARNTL2 expression and tumor immune microenvironment (TIME).ResultsARNTL2 was significantly upregulated in ccRCC tissues and cell lines compared to normal kidney tissues and cell line. Enhanced expression of ARNTL2 was strongly linked to advanced clinical stage and unfavorable overall survival in ccRCC. ARNTL2 was determined as an independent prognostic marker through cox regression analysis. A prognostic nomogram was constructed to predict 1-, 3- and 5-year overall survival of ccRCC patients by integrating ARNTL2 expression with other clinicopathologic variables. GSEA analysis showed that focal adhesion, T cell receptor, cell cycle, and JAK-STAT signaling pathway were significantly enriched in high ARNTL2 samples. Silencing of ARNTL2 suppressed the colony formation ability and wound healing efficacy of ccRCC cell lines. xCell analysis showed that high expression level of ARNTL2 exhibited an immune infiltration status similar to CD8 + inflamed ccRCC subtype, which was characterized by high infiltration level of CD8 + T cell and high expression level of the immune escape biomarkers such as PD-L1, PD-L2, PD1 and CTLA4.ConclusionARNTL2 is an independent adverse predictor of ccRCC patient survival. High expression level of ARNTL2 is associated with immune infiltration, and may be a novel therapeutic target in ccRCC.
Highlights
Aryl hydrocarbon receptor nuclear translocator like 2 (ARNTL2) is a member of the PAS superfamily
ARNTL2 is highly expressed in ccRCCThe transcriptional expression level of ARNTL2 in pan-cancer was preliminarily investigated by analyzing the RNA-seq from The cancer genome atlas (TCGA) and Genotype-tissue expression (GTEx) database
Three independent datasets from Gene expression omnibus (GEO) database were utilized to externally illustrate the expression of ARNTL2 in Clear cell renal cell carcinoma (ccRCC), which demonstrated the highly expressed of ARNTL2 at the transcriptional level in ccRCC (Fig. 1d, e), and increased expression of ARNTL2 significantly associated with the advanced ccRCC clinical stage (Fig. 1f, Table 1) and tumor histologic grade (Table 1), while not significantly associated with age and gender
Summary
Aryl hydrocarbon receptor nuclear translocator like 2 (ARNTL2) is a member of the PAS superfamily. This study sought to explore the roles of ARNTL2 in ccRCC and determine its correlations with tumor immunity. Studies reported that interactions between cancer cells and immune system play a vital role in carcinogenesis of ccRCC [4, 5]. Current immunotherapies, such as monoclonal antibodies targeting PD-L1 and/or CTLA4, have become the main therapy for advanced ccRCC treatment [6]. A series of clinical trials demonstrated that Nivolumab in combination with Ipililmumab was superior to targeted therapy in patients with advanced ccRCC [8, 9], ushering in a new era of first-line treatment for advanced kidney cancer. There is need to discover effective prognostic biomarkers and highly specific tumor immune related therapeutic targets to improve treatment of ccRCC patients
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
