title: 4-methylumbelliferone inhibits hepatocellular carcinoma growth by decreasing IL-6 production and angiogenesis.
abstract: Cirrhosis is characterized by an excessive accumulation of extracellular matrix components including hyaluronic acid (HA) and is widely considered a preneoplastic condition for hepatocellular carcinoma (HCC). 4-Methylumbelliferone (4MU) is an inhibitor of HA synthesis and has anticancer activity in an orthotopic HCC model with underlying fibrosis. Our aim was to explore the effects of HA inhibition by 4MU orally administered on tumor microenvironment. Hepa129 tumor cells were inoculated orthotopically in C3H/HeJ male mice with fibrosis induced by thioacetamide. Mice were orally treated with 4MU. The effects of 4MU on angiogenesis were evaluated by immunostaining of CD31 and quantification of proangiogenic factors (vascular endothelial growth factor, VEGF, interleukin-6, IL-6 and C-X-C motif chemokine 12, CXCL12). IL-6 was also quantified in Hepa129 cells in vitro after treatment with 4MU. Migration of endothelial cells and tube formation were also analyzed. As a result, 4MU treatment decreases tumor growth and increased animal survival. Systemic levels of VEGF were significantly inhibited in 4MU-treated mice. Expression of CD31 was reduced after 4MU therapy in liver parenchyma in comparison with control group. In addition, mRNA expression and protein levels of IL-6 and VEGF were inhibited both in tumor tissue and in nontumoral liver parenchyma. Interestingly, IL-6 production was dramatically reduced in Kupffer cells isolated from 4MU-treated mice, and in Hepa129 cells in vitro. Besides, 4MU was able to inhibit endothelial cell migration and tube formation. In conclusion, 4MU has antitumor activity in vivo and its mechanisms of action involve an inhibition of angiogenesis and IL-6 production. 4MU is an orally available molecule with potential for HCC treatment.
authors.affiliation: Liver Unit, Gene Therapy Laboratory, Facultad de Ciencias Biomédicas, Universidad Austral, Avenida Presidente Perón 1500 (B1629ODT), Derqui-Pilar, Buenos Aires, Argentina.
authors.forename: Flavia
authors.initials: F
authors.lastname: Piccioni
authors.affiliation: Liver Unit, Gene Therapy Laboratory, Facultad de Ciencias Biomédicas, Universidad Austral, Avenida Presidente Perón 1500 (B1629ODT), Derqui-Pilar, Buenos Aires, Argentina.
authors.forename: Esteban
authors.initials: E
authors.lastname: Fiore
authors.affiliation: Liver Unit, Gene Therapy Laboratory, Facultad de Ciencias Biomédicas, Universidad Austral, Avenida Presidente Perón 1500 (B1629ODT), Derqui-Pilar, Buenos Aires, Argentina.
authors.forename: Juan
authors.initials: J
authors.lastname: Bayo
authors.affiliation: Liver Unit, Gene Therapy Laboratory, Facultad de Ciencias Biomédicas, Universidad Austral, Avenida Presidente Perón 1500 (B1629ODT), Derqui-Pilar, Buenos Aires, Argentina CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas), Buenos Aires, Argentina.
authors.forename: Catalina
authors.initials: C
authors.lastname: Atorrasagasti
authors.affiliation: Liver Unit, Gene Therapy Laboratory, Facultad de Ciencias Biomédicas, Universidad Austral, Avenida Presidente Perón 1500 (B1629ODT), Derqui-Pilar, Buenos Aires, Argentina.
authors.forename: Estanislao
authors.initials: E
authors.lastname: Peixoto
authors.affiliation: Liver Unit, Gene Therapy Laboratory, Facultad de Ciencias Biomédicas, Universidad Austral, Avenida Presidente Perón 1500 (B1629ODT), Derqui-Pilar, Buenos Aires, Argentina.
authors.forename: Manglio
authors.initials: M
authors.lastname: Rizzo
authors.affiliation: Liver Unit, Gene Therapy Laboratory, Facultad de Ciencias Biomédicas, Universidad Austral, Avenida Presidente Perón 1500 (B1629ODT), Derqui-Pilar, Buenos Aires, Argentina CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas), Buenos Aires, Argentina.
authors.forename: Mariana
authors.initials: M
authors.lastname: Malvicini
authors.affiliation: Laboratory of Reproductive Medicine, Medicine University of Berlin, Charité Centre 12 Internal Medicine and Dermatology, Berlin, Germany.
authors.forename: Irene
authors.initials: I
authors.lastname: Tirado-González
authors.affiliation: Liver Unit, Gene Therapy Laboratory, Facultad de Ciencias Biomédicas, Universidad Austral, Avenida Presidente Perón 1500 (B1629ODT), Derqui-Pilar, Buenos Aires, Argentina CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas), Buenos Aires, Argentina.
authors.forename: Mariana G
authors.initials: MG
authors.lastname: García
authors.affiliation: CIT NOBA, Universidad Nacional del Noroeste de la Pcia. de Bs. As., Junín, Buenos Aires, Argentina CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas), Buenos Aires, Argentina gmazzoli@cas.austral.edu.ar laualaniz@yahoo.com.ar.
authors.forename: Laura
authors.initials: L
authors.lastname: Alaniz
authors.affiliation: Liver Unit, Gene Therapy Laboratory, Facultad de Ciencias Biomédicas, Universidad Austral, Avenida Presidente Perón 1500 (B1629ODT), Derqui-Pilar, Buenos Aires, Argentina CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas), Buenos Aires, Argentina gmazzoli@cas.austral.edu.ar laualaniz@yahoo.com.ar.
authors.forename: Guillermo
authors.initials: G
authors.lastname: Mazzolini
mesh_terms: D000284:Administration, Oral; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000235:genetics / Q000401:mortality / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D054377:Chemokine CXCL12 / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D006923:Hymecromone / Q000494:pharmacology*; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D007728:Kupffer Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008103:Liver Cirrhosis / Q000139:chemically induced / Q000188:drug therapy* / Q000235:genetics / Q000401:mortality; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000401:mortality / Q000473:pathology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D016019:Survival Analysis; D013853:Thioacetamide; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000093242:Interleukin-6 Inhibitors
chemical_list: D000970:Antineoplastic Agents; D054377:Chemokine CXCL12; D006923:Hymecromone; D015850:Interleukin-6; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D013853:Thioacetamide; D042461:Vascular Endothelial Growth Factor A; D000093242:Interleukin-6 Inhibitors; C515256:Cxcl12 protein, mouse; C467480:vascular endothelial growth factor A, mouse
keywords: 4-methylumbelliferone; HCC; IL-6; VEGF; angiogenesis; hyaluronic acid; liver fibrosis