Effect of extracellular environmental factors in cholangiocarcinoma cells invasiveness

Abstract

The function of the extracellular matrix (ECM) in the tumor microenvironment is not limited to forming a barrier against tumor invasion. As demonstrated in pathological specimens, cholangiocarcinoma samples display an enrichment of the ECM surrounding the tumor cells. In this study, we examined the role that the ECM plays in the regulation of the invasiveness of cholangiocarcinoma cells. The cholangiocarcinoma cell line RMCCA1 was cultured in culture plates either with or without a coating of reconstituted ECM basement membrane preparation (BD Matrigel matrix). In vitro invasion assays were then performed. In addition, the protein expression profile of the cell line was examined using two-dimensional (2D) gel electrophoresis and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The proteins expressed and their functional associations with cancer progression were determined. Culturing the RMCCA1 cell line in the BD Matrigel matrix induced cell invasion. Many proteins were induced by culturing the RMCCA1 cells in the matrix gel. The expression of L-plastin, an actin-binding protein, and pyruvate kinase M2 (PKM2), responsible for energy production within the glycolytic pathway, were significantly upregulated. The knockdown of L-plastin and PKM2 expression by siRNA silencing significantly suppressed the cellular response to matrix gel-stimulated cancer cell invasion. The ECM promotes the invasiveness of cholangiocarcinoma cells by upregulating L-plastin and PKM2. These findings suggest the potential exploitation of this mechanism as a means of inhibiting the invasiveness of cholangiocarcinoma cells.