Hyaluronan modulates tumor progression, angiogenesis and proliferation during Doxorubicin treatment

VITALE DAIANA; SPINELLI FIORELLA; CAON I; DEMARCHI G; PASSI A; CRISTINA C; ALANIZ L

Workshop; 6th FEBS Advanced Lecture Course Matrix Pathobiology, Signaling and Molecular Targets; 2017

The microenvironment generated around the tumor can modulate chemotherapy response. Extracellular matrix (ECM) deregulation could be crucial for tumor progression and metastasis. Hyaluronan (HA), a ECM glycosaminoglycan, interact with its principal receptor (CD44) and induces signals that can promote cell proliferation, angiogenesis, and could be related with apoptosis and multidrug resistance. The objective of our study is to analyze the mechanisms triggered by HA through the interaction with CD44: modulation of signaling pathways associated with angiogenesis and drug resistance.EL4 lymphoma T murine cell line was treated with low molecular weight HA (LMW HA 20-100 μg/ml) and Doxorubicin (DOX 0,5-1-2,5 μM). CD44 expression and HA binding capability were measured by flow cytometry. HA effect during DOX treatment was studied analyzing cell survival, angiogenesis and intracellular signaling. Apoptosis (by AnnexinV) and DOX accumulation assays were performed by flow citometry. Angiogenesis modulation was studied evaluating VEGF expression by ELISA assay and endothelial cells (EC) migration through wound healing assay. To analyze Wnt/β-Catenin pathway, Wnt ligand (β-Catenin) total expression was determined by Western Blot. Three independent experiments were performed for each test, analyzed by one-way or two-way ANOVA considering a significant p value