title: Hepatocyte growth factor and inducible nitric oxide synthase are involved in multidrug resistance-induced angiogenesis in hepatocellular carcinoma cell lines.
abstract: Based on literature, it is possible to hypothesize that multidrug resistance (MDR) and angiogenic phenotypes are linked to each other in human liver cancer cells. Our goal is to assess whether MDR cells trigger angiogenesis and to study the possible molecular mechanisms involved. Conditioned medium from parental drug-sensitive P5 cells (P5-CM) and MDR-positive P1(0.5) cells [P1(0.5)-CM] stimulated human umbilical vein endothelial cells (HUVEC) survival, proliferation, migration, and microtubular structure formation, but P1(0.5)-CM had a significantly greater effect than P5-CM. Cell implants were done in the rabbit avascular cornea to measure angiogenesis in vivo: P1(0.5) cells induced an important neovascular response in rabbit cornea after 1 week, whereas P5 cells had no effect. P1(0.5) and P5 cells produced vascular endothelial growth factor, but only P1(0.5) secreted hepatocyte growth factor (HGF) into the medium, and small interfering RNA specific for MDR1 clearly reduced HGF production in P1(0.5) cells. The transcription factor Ets-1 and the HGF receptor c-Met were up-regulated in P1(0.5) cells and in HUVEC cultured in P1(0.5)-CM. Inducible nitric oxide synthase (iNOS) seemed to play a major role in the proangiogenic effect of P1(0.5), and its inhibition by 1400W blunted the capacity of P1(0.5) cells to stimulate HUVEC proliferation, migration, and Ets-1 expression. In conclusion, these data show that development of MDR and angiogenic phenotypes are linked to each other in MDR cells. HGF production, Ets-1 and c-Met up-regulation, and iNOS expression can be part of the molecular mechanisms that enhance the angiogenic activity of the MDR-positive hepatocellular carcinoma cell line.
authors.affiliation: Department of Internal Medicine, Postgraduate School in Oncology, DENOthe, University of Florence, Azienda Ospedaliero-Universitaria Careggi, Florence, Italy.
authors.forename: Nadia
authors.initials: N
authors.lastname: Lasagna
authors.forename: Ornella
authors.initials: O
authors.lastname: Fantappiè
authors.forename: Michela
authors.initials: M
authors.lastname: Solazzo
authors.forename: Lucia
authors.initials: L
authors.lastname: Morbidelli
authors.forename: Serena
authors.initials: S
authors.lastname: Marchetti
authors.forename: Greta
authors.initials: G
authors.lastname: Cipriani
authors.forename: Marina
authors.initials: M
authors.lastname: Ziche
authors.forename: Roberto
authors.initials: R
authors.lastname: Mazzanti
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000098:blood supply* / Q000201:enzymology; D002450:Cell Communication / Q000502:physiology; D048708:Cell Growth Processes / Q000502:physiology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D016510:Corneal Neovascularization; D051546:Cyclooxygenase 2 / Q000096:biosynthesis; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis / Q000502:physiology*; D006801:Humans; D008113:Liver Neoplasms / Q000098:blood supply* / Q000201:enzymology; D009389:Neovascularization, Pathologic / Q000201:enzymology / Q000473:pathology; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism / Q000502:physiology*; D050784:Proto-Oncogene Protein c-ets-1 / Q000096:biosynthesis; D019859:Proto-Oncogene Proteins c-met / Q000096:biosynthesis; D034622:RNA Interference; D011817:Rabbits; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis
chemical_list: D050784:Proto-Oncogene Protein c-ets-1; D042461:Vascular Endothelial Growth Factor A; D017228:Hepatocyte Growth Factor; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2; D019859:Proto-Oncogene Proteins c-met