Differential expression and prognostic significance of ANKRD6 in colon adenocarcinoma: a pan-cancer analysis and pathway-specific investigation

Abstract

Aim: This study aims to investigate the expression of the Ankyrin Repeat Domain 6 (ANKRD6) gene in Colon Adenocarcinoma (COAD) and its regulatory role in key signaling pathways, evaluating its clinical value as a potential therapeutic target.

Methods: To investigate the functional role of ANKRD6 in colon adenocarcinoma (COAD), we systematically analyzed its prognostic relevance, epigenetic regulation, and association with tumor stemness using publicly available TCGA (The Cancer Genome Atlas)-COAD data. Gene set variation analysis (GSVA) was applied to identify signaling pathways potentially modulated by ANKRD6. Least Absolute Shrinkage and Selection Operator (LASSO) regression was employed to identify key genes within the Wnt signaling pathway that are significantly associated with COAD patient survival. Immunohistochemical staining was performed to confirm ANKRD6 protein expression in COAD tissues. To investigate functional consequences, ANKRD6 was knocked down using small interfering RNAs, and subsequent quantitative real-time polymerase chain reaction and Western blot assays were applied to measure alterations in representative Wnt pathway-related genes and proteins.

Results: ANKRD6 expression was significantly associated with DNA methylation patterns, RNA modification regulators, and tumor stemness features in COAD. These findings suggest that epigenetic and post-transcriptional mechanisms may underlie the regulatory role of ANKRD6 in COAD pathogenesis. GSVA analysis revealed that ANKRD6 significantly influences and negatively regulates Wnt signaling pathway activity. LASSO regression analysis highlighted 25 Wnt pathway genes relevant to COAD patient survival, with ANKRD6 expression significantly correlated with multiple targets. Furthermore, immunohistochemical staining confirmed that ANKRD6 protein was markedly upregulated in COAD tissues. Upon ANKRD6 knockdown, the messenger RNA levels of WNT7A (Wnt Family Member 7A), JNK (Mitogen-Activated Protein Kinase 8), and RHOA (Ras Homolog Family Member A), as well as WNT7A protein levels, showed a significant decrease in the Wnt signaling pathway.

Conclusions: ANKRD6 plays a critical role in the development and progression of COAD by regulating the Wnt signaling pathway. It holds potential as a therapeutic target, warranting further investigation into its specific regulatory mechanisms and clinical applications.

Keywords

ANKRD6 / COAD / DNA methylation / pan-cancer analysis / tumor stemness / GSVA / LASSO regression analysis

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