Abstract 2146 Studying Cellular and Protein Dynamics of Cultured Cell Lines Exposed to Tubulin Disruptors, Bisphenol-A and Taxol

The presence of tubulin disruptors or tubulin-targeting agents, like Taxol and Colchicine are used to help treat diseases like cancer and gout, respectively. Both types of chemicals work either by polymerization or depolymerization of microtubules arresting the cell or causing cell death. Previous studies have shown that Bisphenol-A (BPA), a weakly estrogenic compound can affect tubulin structures in oocytes, early embryonic development, and cell cultures by displaying disruptions in the spindle apparatus during cell division in mitosis and meiosis. This study intends to use both biochemistry and cellular microscopy to analyze and characterize spindle assembly proteins and cytoskeletal proteins that are expressed in cancerous and non-cancerous cells when exposed to BPA compared to other known tubulin disruptors, like Taxol. There will also be focused attention on understanding the different binding mechanisms that these tubulin disruptors have to the three different types of tubulin proteins, α-, β-, and γ-tubulin. We conducted this study by growing and synchronizing cell cultures, HEK-293 (non-cancerous) and MCF7 (cancerous) cell lines, in both control and experimental groups. After cells were released from the thymidine block, they were exposed to BPA or Taxol during M phase and samples were collected every two hours. We fixed samples on coverslips and collected cells for protein analysis. Fixed samples will be stained with fluorescence using antibodies against the three tubulin proteins and imaged using confocal microscopy. Protein samples will also be analyzed by SDS Page and Western Blot to confirm the disruptions of the tubulin proteins. The application of the information obtained by this study will allow other researchers to further understand the mechanisms of how different tubulin disruptors can bind to cytoskeletal proteins and how it can be used to improve treatments in cancer cells in future studies. This project was supported by grants from the National Institutes of Health (NIH), National Institute of General Medical Sciences (NIGMS), IDeA Networks of Biomedical Research Excellence (INBRE), Award number: . The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.