INTERNATIONAL CONGRESS OF
CancerGenetics
Targeting DYRK1B in triple-negative breast cancer: CRISPR-Cas9 mediated knockout reveals potential for therapeutic intervention
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Asrin Rashidi,1 Fardin Fathi,2,* Zakaria Vahabzadeh,3 Farzad Soleymani,4 Asaad Azarnezhad,5 Arash Pooladi,6
1. Department of Molecular Medicine, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran
2. Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran
3. Department of Biochemistry, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran
4. Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran
5. Department of Molecular Genetic, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran
6. Department of Molecular Genetic, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran
- Introduction: Breast cancer is the most common cancer affecting women worldwide, with the triple-negative subtype (TNBC) being particularly aggressive and difficult to treat. DYRK1B, a dual-specificity kinase, is known to regulate cell cycle and quiescence. Although its role in various cancers has been studied, its impact on TNBC has not been well understood.
- Methods: In this study, we employed CRISPR-Cas9 technology to knock out DYRK1B in MDA-MB-231 cells, a model for TNBC. We then assessed the effects on cell proliferation, apoptosis, invasion, migration, angiogenesis, and response to the chemotherapy drug Paclitaxel.
- Results: The successful knockout (KO) of DYRK1B was confirmed using PCR, Sanger sequencing, and real-time qPCR. Compared to wild-type (WT) MDA-MB-231 cells, KO cells showed a marked decrease in cell proliferation, colony formation, migration, and invasion. Additionally, KO cells exhibited 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 CCND1, MCM2, PCNA, CDKN1B, HIF1A, VEGFA, and WASF3. Immunocytochemistry further demonstrated that KO cells had significantly lower Ki67 expression, indicating reduced cell proliferation.
- Conclusion: Our findings highlight the critical role of DYRK1B in the survival and invasive capabilities of TNBC cells. Targeting DYRK1B could offer a promising new therapeutic strategy for treating this challenging subtype of breast cancer. These results underscore the potential of DYRK1B as a novel target for therapeutic intervention in TNBC, paving the way for more effective treatments.
- Keywords: Brest cancer, CRISPR-Cas9, DYRK1B, Quiescent cancer cell, Targeted therapy
