Upregulation of ARNTL2 is associated with poor survival and immune infiltration in clear cell renal cell carcinoma

Abstract

Background Aryl 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. Methods The 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). Results ARNTL2 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. Conclusion ARNTL2 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.

ARNTL2 is a Prognostic Biomarker and Correlates with Immune Cell Infiltration in Triple-Negative Breast Cancer

“…Moreover, a recent study demonstrated that high expression of ARNTL2 was associated with infiltration of CD8 + T cell, CD4 + memory T cell, Myeloid dendritic cell, macrophage and CD4 + Th2 T cell in clear cell renal cell carcinoma (ccRCC). 43 Thus, the results of the present study and others demonstrated that ARNTL2 may functionally link with immune responses in the tumor microenvironment and represent a potential indicator for immune cell infiltration.…”

Targeting the ARNTL2 Gene as a Potential Strategy for Lung Adenocarcinoma

“…As an important biorhythm-related gene, ARNTL has been reported to play an oncogenic role in many human cancers. For example, Prior studies found that upregulation of ARNTL2 is associated with poor survival and immune in ltration in clear cell renal cell carcinoma 15 . Mazzoccoli et al reported that ARNTL2 is upregulated in colorectal cancer and is related to tumor invasiveness and aggressiveness 16 .…”

Prognostic Value of Long Noncoding RNA DLEU2 and Its Relationship with Immune Infiltration in Kidney Renal Clear Cell Carcinoma and Liver Hepatocellular Carcinoma

“… 7 , 8 Previous studies have reported that the prognosis of KIRC is closely related to immune-infiltrating macrophages, CD4+ T cells, CD8+ T cells, and regulatory T cells. 9 , 10 The treatment outcome of LIHC is believed to be correlated with infiltrating macrophages, B cells, CD4+ T cells, neutrophils, and dendritic cells. 11 The immunoregulatory molecules PD1, PD-L1, PD-L2, and CTLA4 are believed to be involved in the malignant progression of KIRC, 9 and PD-1 and CTLA-4 are believed to be associated with a poor prognosis of LIHC.…”

“… 9 , 10 The treatment outcome of LIHC is believed to be correlated with infiltrating macrophages, B cells, CD4+ T cells, neutrophils, and dendritic cells. 11 The immunoregulatory molecules PD1, PD-L1, PD-L2, and CTLA4 are believed to be involved in the malignant progression of KIRC, 9 and PD-1 and CTLA-4 are believed to be associated with a poor prognosis of LIHC. 12 These findings suggest that tumour-infiltrating immune cells may be a promising therapeutic target for KIRC and LIHC.…”