Transgelin Depletion is Critical for the TGF beta 1-mediated Initiation of PLC gamma 1-Cofilin-driven Morphological and Migratory Changes in MDA-MB-231 Cells

Abstract

The role of transgelin (TAGLN) in cancer has been discussed; however, the mechanisms underlying its regulation and correlation with MDA-MB-231 cell plasticity and migratory patterns remain unclear. We generated stable TAGLN-knockdown MDA-MB-231 cells and treated them with phorbol 12-myristate 13-acetate or transforming growth factor (TGF)-beta. Chemotaxis, morphology, and invasion were assayed using three-dimensional matrices to evaluate cytoskeletal remodeling and migratory changes. Wound healing assays were conducted using cell inserts. TAGLN knockdown cells exhibited altered morphology due to cytoskeletal remodeling, yet only untreated and TGF beta 1-treated cells exhibited enhanced migration. Untreated and TGF beta 1-treated TAGLN knockdown cells showed increased N-WASP, ROCK1, and ROCK2 protein levels, which induce cytoskeletal remodeling. Evaluating phospholipase C gamma 1 (PLC gamma 1)-cofilin signaling-related proteins revealed that only TGF beta 1-treated TAGLN knockdown cells were influenced by PLC gamma 1-cofilin signaling. Taken together, TAGLN knockdown is necessary for the TGF beta 1-mediated activation of PLC gamma 1-cofilin pathway-driven amoeboid morphology and enhanced migratory properties in MDA-MB-231 cells.