The role of Rho GTPase in cell stiffness and cisplatin resistance in ovarian cancer cells

Abstract

Changes in cell stiffness (Young's modulus, E), as measured via Atomic Force Microscopy (AFM), is a newly recognized characteristic of cancer cells and may play a role in platinum drug resistance of ovarian cancers. We previously showed that, compared to their syngeneic cisplatin-sensitive counterpart, cisplatin-resistant ovarian cancer cells are stiffer, and this cell stiffness was dependent on actin polymerization and presence of stress fibers. Here, we measured the correlation between Young's modulus (via AFM measurements on live, non-apoptotic cells in physiological buffer) and cisplatin-sensitivity (IC₅₀ as determined via the XTT cell viability assay) in a panel of nine ovarian cancer cell lines representing a range of cisplatin sensitivities. We found that cisplatin-sensitive cells had a lower Young's modulus, compared to cisplatin-resistant cells and resistant cells had a cytoskeleton composed of long actin stress fibers. As Rho GTPase mediates stress fiber formation, we examined the role of Rho GTPase in cell stiffness and platinum resistance. Rho inhibition decreased cell stiffness in cisplatin-resistant CP70 cells and increased their cisplatin sensitivity while Rho activation increased cell stiffness in cisplatin-sensitive A2780 cells and decreased their cisplatin sensitivity. Based on changes in cell stiffness, IC₅₀ and cellular actin stress fiber organization in CP70 and A2780 cells, our findings reveal a direct role of Rho mediated actin remodeling mechanism in cisplatin resistance of ovarian cancer cells. These findings suggest the potential applicability of cell mechanical phenotyping as a model for determining sensitivity of ovarian cancer cells that could have major implications in ovarian cancer diagnosis and personalized medicine.