4.7 Article

Ginkgo biloba exocarp extracts inhibits angiogenesis and its effects on Wnt/β-catenin-VEGF signaling pathway in Lewis lung cancer

Journal

JOURNAL OF ETHNOPHARMACOLOGY
Volume 192, Issue -, Pages 406-412

Publisher

ELSEVIER IRELAND LTD
DOI: 10.1016/j.jep.2016.09.018

Keywords

Ginkgo biloba exocarp extracts; Wnt3a; beta-catenin; VEGF; VEGFR2; Angiogenesis; Lewis lung cancer cells

Funding

  1. Medical High-tech Research Projects of Jiangsu Province [BG2007609]
  2. science and technology innovation fund projects of college students in Yangzhou University [x2015767]

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Ethnopharmacological relevance: 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. Aim of study: Study on the activity of GBEE against Lewis lung cancer (LLC) angiogenesis and its partial molecular mechanism. Materials and 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, 200 mg/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 0.1 mL/10 g (b.w.), positive group were intraperitoneal (i.p.) injection cyclophosphamide (CPA) at a dose of 20 mg/kg (b.w.), the GBEE treatment groups were respectively i.g. GBEE 50, 100, and 200 mg/kg (b.w.), once a day for 20 d. 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, beta-catenin, VEGF, VEGFR2 and beta-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 160 mu g/mL in vitro. It can suppressed Wnt3a and P-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, 200 mg/kg, suppressing tumor lung metastasis. The expression of CD34 was reduced, which means MVD was inhibited and so do beta-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 /beta-catenin-VEGF signaling pathway in LLC. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

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