Quantification of Invadopodia Formation and Matrix Degradation Activity

Abstract

Cancer cells possess a remarkable capacity to dissociate from a primary tumor, invade the surrounding tissues and vasculature, and eventually form metastases in distant organs. This complex and multistep process remains one of the major causes of mortality in cancer patients worldwide. Multiple studies have highlighted the role of actin-rich structures called invadopodia ("invasive feet"), which adhere to the matrix, contain and secrete matrix-degrading proteinases, and apply protrusive forces generated by the actin cytoskeleton, which drive the invasive process. Here, we describe a fluorescent microscopy-based protocol for imaging and quantifying both invadopodia formation and matrix degradation.

Keywords: Actin cytoskeleton; Cancer cell lines; Extracellular matrix (ECM); Fluorescent microscopy; Fluorescently labeled gelatin; Gelatin degradation; Image processing; Invadopodia; Invasion; Metastasis.