Mitosis, Cytoskeleton Regulation, and Drug Resistance in Receptor Triple Negative Breast Cancer

Abstract

During cell division, the receptor triple-negative MDA-MB-231 mitotic spindles are the largest in comparison to other BC cell lines. Many of the MDA-MB-231 spindles exhibit rapid lateral twisting during metaphase, which remains unaffected by knockdown of the oncogene Myc and treatment with inhibitors of the serine/threonine-protein kinase B-Raf and the epidermal growth factor receptor (EGFR), alone or in any combination. The MDA-MB-231 cells are the most aggressive and rapidly form metastatic tumors in xenograft transplant models, and exhibited very high proliferation rates when plated as three-dimensional cultures in Matrigel. Quantitative image analysis of microtubules (MTs) in six BC cell lines - MDA-MB-231 (receptor negative), HCC-1143 (receptor negative), HCC-3153 (receptor negative), ZR75B (estrogen receptor-positive), LY2 (progesterone receptor-positive), HCC-1428 (estrogen receptor-positive, progesterone receptor-positive) - demonstrated that the rotational spindle rocking of MDA-MB-231 cells during metaphase appears coupled with a significant increase in MT polymerization rates during interphase, which likely shortens interphase and accelerates cell cycle progression and mitotic entry. Unlike the uniform treadmilling rates of about 21 um/min in kinetochore MTs during metaphase we measured across cell lines, MDA-MB-231 cells in interphase exhibit the fastest MT polymerization dynamics of about 19 um/min and this is coupled with abnormal mitotic spindle oscillations of almost 30 um/min. This aberrant behavior in MDA-MB-231 spindles may represent a therapeutically targetable disrupted mechanism of spindle positioning in receptor triple-negative breast cancer (TNBC) cells leading to tumor aggressiveness. In this manuscript, we outline a strategy for the selection of the most optimal tubulin inhibitor based on the ability to affect MT dynamics.