Targeting DYRK۱B in triple-negative breast cancer: CRISPR-Cas۹ mediated knockout reveals potential for therapeutic intervention

Introduction: Breast cancer is the most common cancer affecting women worldwide, with the triple-negative subtype (TNBC) being particularly aggressive and challenging to treat. DYRK۱B, a dual-specificity kinase, plays a role in regulating the cell cycle and cellular quiescence. While its function in various cancers has been studied, its impact on TNBC remains poorly understood.Methods: In this study, we utilized CRISPR-Cas۹ technology to knock out DYRK۱B in MDA-MB-۲۳۱ cells, a model for TNBC. We assessed the effects of this knockout on cell proliferation, apoptosis, invasion, migration, angiogenesis, and response to the chemotherapy drug Paclitaxel.Results: The successful knockout of DYRK۱B was confirmed through PCR, Sanger sequencing, and real-time qPCR. Compared to wild-type (WT) MDA-MB-۲۳۱ cells, DYRK۱B knockout (KO) cells exhibited a marked decrease in cell proliferation, colony formation, migration, and invasion. Additionally, KO cells showed increased apoptosis and heightened sensitivity to contact inhibition and Paclitaxel. Gene expression analysis revealed altered levels of several genes associated with the cell cycle, angiogenesis, and cell motility, including CCND۱, MCM۲, PCNA, CDKN۱B, HIF۱A, VEGFA, and WASF۳. Immunocytochemistry further indicated significantly lower Ki۶۷ expression in KO cells, reflecting reduced cell proliferation.Conclusion: Our findings highlight the critical role of DYRK۱B in the survival and invasive capabilities of TNBC cells. Targeting DYRK۱B may provide a promising therapeutic strategy for treating this challenging breast cancer subtype. These results underscore the potential of DYRK۱B as a novel target for therapeutic intervention in TNBC, paving the way for more effective treatments.

کلیدواژه ها:

Apoptosis ، Breast cancer ، CRISPR-Cas۹ ، DYRK۱B ، Quiescent cancer cell ، Targeted therapy