title: Ginkgo biloba exocarp extracts inhibits angiogenesis and its effects on Wnt/β-catenin-VEGF signaling pathway in Lewis lung cancer.
abstract: BACKGROUND
  A fruit of Ginkgo biloba L. also known as Ginkgo biloba, can be used for the treatment of cancer in Chinese traditional medicine. The scientific name of succulent skin, which is the episperm of Ginkgo nuts, is exocarp. Experiment shows that Ginkgo biloba exocarp extracts (GBEE) has the effects of immune promotion, cancer inhibition and etc.
  
  
  OBJECTIVE
  Study on the activity of GBEE against Lewis lung cancer (LLC) angiogenesis and its partial molecular mechanism.
  
  
  METHODS
  The effect of GBEE on proliferation of LLC cells was detected by MTT method in vitro. The metastasis model of LLC was set up. The C57BL/6J mice were randomly separated into normal control, model control, positive control and GBEE (50, 100, 200mg/kg) treatment groups, n=10. The mice in normal group and model group were both intragastric gavage (i.g.) normal saline (NS) in a volume of 0.1mL/10g (b.w.), positive group were intraperitoneal (i.p.) injection cyclophosphamide (CPA) at a dose of 20mg/kg (b.w.) , the GBEE treatment groups were respectively i.g. GBEE 50, 100, and 200mg/kg (b.w.), once a day for 20d. After treatment, we calculated the tumor inhibition rate and anti-metastasis rate. The microvessel density (MVD) was measured by immunohistochemistry method in transplanted tumor. The expression levels of vascular en-dothelial growth factor (VEGF) and VEGFR2 mRNA or Wnt3a, β-catenin, VEGF, VEGFR2 and p-Akt/Akt protein expression were respectively tested by Quantitative Reverse transcription Polymerase chain reaction (qRT-PCR) or western blot in vitro and vivo.
  
  
  RESULTS
  GBEE suppressed the growth of LLC cells in a dose-dependent way at the dose of 5, 10, 20, 40, 80 and 160µg/mL in vitro. It can suppressed Wnt3a and β-catenin protein expression and the content of mRNA of VEGF and VEGFR2 in LLC cells significantly. In vivo, we discovered GBEE can retard the growth of LLC transplanted tumor in a dose-dependent way at the dose of 50, 100, 200mg/kg, suppressing tumor lung metastasis. The expression of CD34 was reduced, which means MVD was inhibited and so do β-catenin, VEGF, VEGFR2 and p-AKT/AKT protein expression and VEGF and VEGFR2 mRNA expression levels in LLC transplanted tumor of C57BL/6 mice.
  
  
  CONCLUSIONS
  GBEE played the effects of anti-tumor and anti-metastatic depending upon the inhibition of tumor angiogenesis, which may be closely relevant to its effect in blockage of Wnt /β-catenin-VEGF signaling pathway in LLC.
authors.affiliation: Department of Pharmacology, Medical College, Yangzhou University, Yangzhou 225001, Jiangsu, China.
authors.forename: Dongdong
authors.initials: D
authors.lastname: Han
authors.affiliation: Department of Pharmacology, Medical College, Yangzhou University, Yangzhou 225001, Jiangsu, China.
authors.forename: Chengjie
authors.initials: C
authors.lastname: Cao
authors.affiliation: Department of Pharmacology, Medical College, Yangzhou University, Yangzhou 225001, Jiangsu, China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Su
authors.affiliation: Department of Pharmacology, Medical College, Yangzhou University, Yangzhou 225001, Jiangsu, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Combination of traditional Chinese and Western Medicine, Medical College, Yangzhou University, Yangzhou 225001, Jiangsu, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Sun
authors.affiliation: Department of Combination of traditional Chinese and Western Medicine, Medical College, Yangzhou University, Yangzhou 225001, Jiangsu, China.
authors.forename: Huasheng
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Pharmacology, Medical College, Yangzhou University, Yangzhou 225001, Jiangsu, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, Jiangsu, China. Electronic address: ahxu@yzu.edu.cn.
authors.forename: Aihua
authors.initials: A
authors.lastname: Xu
mesh_terms: D020533:Angiogenesis Inhibitors / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology*; D018827:Carcinoma, Lewis Lung / Q000098:blood supply / Q000188:drug therapy* / Q000378:metabolism / Q000556:secondary; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003520:Cyclophosphamide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D020441:Ginkgo biloba / Q000737:chemistry*; D008810:Mice, Inbred C57BL; D055806:Microvessels / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D009389:Neovascularization, Pathologic; D010766:Phosphorylation; D008517:Phytotherapy; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D010946:Plants, Medicinal; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D047368:Tumor Burden / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects*; D060509:Wnt3A Protein / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism; D000096983:Ginkgo Extract
chemical_list: D020533:Angiogenesis Inhibitors; D000972:Antineoplastic Agents, Phytogenic; D003520:Cyclophosphamide; D010936:Plant Extracts; D051057:Proto-Oncogene Proteins c-akt; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2; D060509:Wnt3A Protein; D051176:beta Catenin; C495267:CTNNB1 protein, mouse; C555511:Wnt3a protein, mouse; C467480:vascular endothelial growth factor A, mouse; C063170:Ginkgo biloba extract; D000096983:Ginkgo Extract; C000592875:Kdr protein, mouse
keywords: Angiogenesis; Arabinose (PubChem CID: 66308); Cis-Dichlorodiammine platinum(II) (PubChem CID: 5702198); Cyclophosphamide (PubChem CID: 2907); Galactose (PubChem CID: 6036); Galacturonic acid (PubChem CID: 439215); Ginkgo biloba exocarp extracts; Glucose (PubChem CID: 5793); Lewis lung cancer cells; Mannose (PubChem CID: 18950); Rhamnose (PubChem CID: 25310); VEGF; VEGFR2; Wnt3a; β-catenin