title: Anti-tumorigenic and anti-angiogenic effects of natural conifer Abies sibirica terpenoids in vivo and in vitro.
abstract: OBJECTIVE
  The natural terpenoid compound was explored in vitro and in vivo to investigate the anti-HCC properties.
  
  
  METHODS
  For our study we used Abisilin®- a novel natural pharmacological terpenoid compound extracted and purified from coniferous Pinaceae trees. Anti-tumorigenic properties of different concentrations of Abisilin®were tested on murine hepatoma Hepa 1-6 cell lines. The analysis of proliferation and apoptosis was performed using immunofluorescence microscopy, FACS and qPCR. As an in vivo approach, we tested Abisilin®(400mg/kg/day, 14 days, orally) in xenograft mouse models of liver cancer and investigated tumor growth, proliferation, apoptosis and angiogenesis by means of Western blotting, immunofluorescence microscopy and qPCR.
  
  
  RESULTS
  Application of Abisilin®for 24h at a dosage ranging from 0.03 to 0.045mg significantly reduced the number of viable Hepa 1-6 cells and induced apoptotic cell death with microscopic evidence of changes in cell morphology, and positive TUNEL, cleaved caspase 3 and Annexin V/Propidium Iodide (PI) stainings. Furthermore, treatment with Abisilin®strongly inhibited proliferation, impaired mitosis and prompted cell cycle arrest by down-regulation of the Cyclin D1, E1 and A2 expression levels. In Hepa 1-6 xenograft in vivo model, Abisilin®considerably decreased the xenograft tumor size and tumor volume. Consistently with in vitro Abisilin®administration elicited apoptosis and inhibit proliferation in the xenograft tumor. We also found that Abisilin®remarkably decrease microvessel density, diminished tumor angiogenesis and reduced expression of ICAM-1. Moreover, the expression of pAMPK, a cellular energy sensor, was up-regulated after Abisilin®application.
  
  
  CONCLUSIONS
  Anti-proliferative, pro-apoptotic activity and anti-angiogenic potential of natural conifer terpenoids might turn these compounds into an attractive drug candidate for combination therapy against liver cancer.
authors.affiliation: Department of Internal Medicine III, University Hospital RWTH Aachen, Pauwelsstrasse 30, D-52074, Aachen, Germany. Electronic address: ynevzorova@ukaachen.de.
authors.forename: Yulia A
authors.initials: YA
authors.lastname: Nevzorova
authors.affiliation: Department of Internal Medicine I, Evangelical Hospital Bethesda, Ludwig-Weber-Strasse 15, 41061, Mönchengladbach, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Grossmann
authors.affiliation: Department of Internal Medicine III, University Hospital RWTH Aachen, Pauwelsstrasse 30, D-52074, Aachen, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Trautwein
mesh_terms: D028202:Abies / Q000737:chemistry*; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D008297:Male; D051379:Mice; D009374:Neoplasms, Experimental / Q000188:drug therapy; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology; D013729:Terpenes / Q000737:chemistry*
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D010936:Plant Extracts; D013729:Terpenes
keywords: Conifer terpenoids; Hepa 1–6 cells; Hepatocellular carcinoma; Xenograft model