TORC2 Mediated Chemotaxis in Mammary Epithelial Cells

Abstract

Chemotaxis is the directional movement of cells in response to an extracellular chemical gradient. It is thought to be involved in cancer cell metastasis by recognizing chemokines and growth factors; therefore, deregulation of chemotactic pathways can result in increased tumor cell metastasis. The Target of Rapamycin Complex 2 (TORC2) regulates chemotaxis downstream of these signaling molecules, or chemoattractants. Examining the role of TORC2 in chemotaxis of cancer cells could provide insight into the deregulation of signals leading to cancer cell metastasis. The non-tumorgenic cell line MCF10A and the two breast cancer cell lines MCF7 and MDA-MB-231 were employed in this study. Epidermal Growth Factor (EGF) and Insulin-like Growth Factor 1 (IGF-1) showed potential as chemoattractants by stimulating TORC2. Wound healing assays were performed on MCF10A, MCF7, and MDA-MB-231 cells exposed to TOR inhibitors, as well as MCF10A cells with Rictor knocked down. Cells with Rictor knocked down and MCF10A cells exposed to Torin 2 showed a decrease in cell migration. Ibidiμ-Slide chemotaxis slides were used to perform a chemotaxis assay with MCF10A cells in response the EGF. Cells showed greater directionality toward EGF in the experimental well as compared to the control with EGF on both sides of the cell chamber. Future examination of other potential chemoattractants as well as chemotaxis assays with other chemoattractants will give more insight into the goals of this research.