Co-treatment of hyaluronan and doxorubicin in tumor cells affects endothelial cell behavior independently of VEGF expression

VITALE, D.L.; SPINELLI, F.M.; DEL DAGO, D.; ICARDI, A.; DEMARCHI, G.; CAON, I.; PASSI, A.; BOLONTRADE, M.F.; CRISTINA, C.; ALANIZ, L.

Oncotarget

Impact Journals, LLC

Lugar: New York; Año: 2018

Hyaluronan is the major glycosaminoglycan component of extracellular matrices, concentrated in tissues with high cell proliferation and migration rates. During the course of cancer, hyaluronan expression is usually altered and it becomes fragmented into low molecular weight forms, affecting mechanisms associated with cell proliferation, invasion, angiogenesis and multidrug resistance. In this work, we analyzed the effect of low molecular weight hyaluronan on doxorubicin efflux and response in lymphoma, osteosarcoma and mammary adenocarcinoma cell lines. Besides, we also analyzed whether tumor cells treated with low molecular weight hyaluronan and doxorubicin differentially modified endothelial cells behavior. After tumor cells? co-treatment by hyaluronan and doxorubicin, we observed that hyaluronan altered drug accumulation and modulated expression of ATP-binding cassette (ABC) transporters in lymphoma cells. Contrasting, neither changes nor an increase in apoptosis were observed when these parameters were analyzed in cells derived from solid tumors, indicating that in this conditions hyaluronan did not affect drug efflux. However, when the effects on angiogenic mechanisms were evaluated, supernatant obtained from tumor cells treated with doxorubicin unexpectedly exhibited a pro-angiogenic effect on endothelial cells. Moreover, we observed that doxorubicin incremented migration and vessel formation ability in co-treatment with hyaluronan. Nevertheless, this effect showed no relation with VEGF levels, which remained similar to basal conditions or even decreased after treatment. It was found that other factors are involved in the angiogenic response, such as FGF-2. On the other hand, we evaluated Wnt/β-catenin and PI3K/Akt pathways as mediators of hyaluronan signals in proliferation and angiogenesis. We observed an increase of β-catenin expression after different LMW HA treatment doses with doxorubicin in all tumor cell lines, indicating that hyaluronan affects doxorubicin treatment. Moreover, we observed higher levels of p-Akt in the MDA-MB-231 cell line, indicanting that hyaluronan in combination with doxorubicin treatment altered this signaling pathway affecting tumor cells elimination. Even though the angiogenic action of hyaluronan in the tumor context is well known, our results demonstrate for the first time that hyaluronan is also a potential modulator of the angiogenic response during doxorubicin treatment, conferring an adverse tumor stroma during chemotherapy.