Inhibition of TNIK with NCB-0846 Reduces Viability and Induces Apoptosis in Triple Negative Breast Cancer Cells

Abstract

Objective: This study aimed to explore the therapeutic potential of NCB-0846, a TNIK inhibitor targeting the Wnt/β-catenin pathway, on triple-negative breast cancer (TNBC) at the molecular level. Materials and Methods: TNBC cell line MDA-MB-231 and non-cancerous breast epithelial cell line MCF-10A were treated with NCB-0846. Cytotoxicity was evaluated using WST-1 assay, apoptosis with Annexin V assay, cell cycle changes with flow cytometry, and morphological alterations with acridine orange staining. RT-PCR was used to assess NCB-0846’s effect on CTNNB1 gene expression. Results: NCB-0846 displayed dose- and time-dependent cytotoxicity in MDA-MB-231 cells, significantly reducing cell viability at 3 μM after 72 hours (p<0.01). Apoptotic cell death was markedly increased at all doses in MDA-MB-231 cells, with notable cell cycle arrest in the G2/M phase (p<0.01). Notably, NCB-0846 induced non-selective toxicity, affecting both cancerous and non-cancerous cells. While CTNNB1 expression decreased in MDA-MB-231 cells, it significantly increased in MCF-10A cells, indicating a potential resistance mechanism in normal breast cells. Conclusion: NCB-0846 reduced TNBC cell viability and induced apoptosis via β-catenin inhibition. However, its lack of selective cytotoxicity may limit its use as a monotherapy. Future research could focus on developing combinational strategies to enhance NCB-0846’s specificity for TNBC, improving its therapeutic potential in clinical settings.

Keywords

• Triple negative breast cancer
• Wnt/β-catenin
• TNIK
• NCB-0846

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