Cytoarchitecture of Breast Cancer Cells under Diabetic Conditions: Role of Regulatory Kinases—Rho Kinase and Focal Adhesion Kinase.

Simple Summary: Diabetes rapidly advances the growth and spread of breast cancer. This study explores how high-glucose conditions in diabetes affect the structure of cancer cells, which makes them more aggressive and more likely to spread to other parts of the body. This is partly due to the involvement of two proteins associated with cytoskeletal structure, Rho kinase (ROCK) and focal adhesion kinase (FAK), which are important for cell movement. Cell shape and stiffness were studied using advanced microscopic techniques in cancer cells and normal cells exposed to varying glucose levels. It was found that cancer levels made the cancer cells less stiff and more likely to move, which can lead to the cancer spreading. However, when the ROCK or FAK proteins were blocked, these harmful changes were lessened. Diabetes greatly reduces the survival rates in breast cancer patients due to chemoresistance and metastasis. Reorganization of the cytoskeleton is crucial to cell migration and metastasis. Regulatory cytoskeletal protein kinases such as the Rho kinase (ROCK) and focal adhesion kinase (FAK) play a key role in cell mobility and have been shown to be affected in cancer. It is hypothesized that diabetes/high-glucose conditions alter the cytoskeletal structure and, thus, the elasticity of breast cancer cells through the ROCK and FAK pathway, which can cause rapid metastasis of cancer. The aim of the study was to investigate the role of potential mediators that affect the morphology of cancer cells in diabetes, thus leading to aggressive cancer. Breast cancer cells (MDA-MB-231 and MCF-7) were treated with 5 mM glucose (low glucose) or 25 mM glucose (high glucose) in the presence of Rho kinase inhibitor (Y-27632, 10 mM) or FAK inhibitor (10 mM). Cell morphology and elasticity were monitored using atomic force microscopy (AFM), and actin staining was performed by fluorescence microscopy. For comparative study, normal mammary breast epithelial cells (MCF-10A) were used. It was observed that high-glucose treatments modified the cytoskeleton of the cells, as observed through AFM and fluorescence microscopy, and significantly reduced the elasticity of the cells. Blocking the ROCK or FAK pathway diminished the high-glucose effects. These changes were more evident in the breast cancer cells as compared to the normal cells. This study improves the knowledge on the cytoarchitecture properties of diabetic breast cancer cells and provides potential pathways that can be targeted to prevent such effects. [ABSTRACT FROM AUTHOR]