Han Dongdong, Cao Chengjie, Su Ya, Wang Jun, Sun Jian, Chen Huasheng, Xu Aihua
Department of Pharmacology, Medical College, Yangzhou University, Yangzhou 225001, Jiangsu, China.
Department of Combination of traditional Chinese and Western Medicine, Medical College, Yangzhou University, Yangzhou 225001, Jiangsu, China.
J Ethnopharmacol. 2016 Nov 4;192:406-412. doi: 10.1016/j.jep.2016.09.018. Epub 2016 Sep 17.
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.
Study on the activity of GBEE against Lewis lung cancer (LLC) angiogenesis and its partial molecular mechanism.
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.
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.
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.
银杏果，也被称为白果，在传统中医中可用于治疗癌症。银杏肉质外皮（即银杏种仁的外种皮）的学名是外种皮。实验表明，银杏外种皮提取物（GBEE）具有免疫促进、抑制癌症等作用。
研究GBEE对Lewis肺癌（LLC）血管生成的活性及其部分分子机制。
采用MTT法体外检测GBEE对LLC细胞增殖的影响。建立LLC转移模型。将C57BL/6J小鼠随机分为正常对照组、模型对照组、阳性对照组和GBEE（50、100、200mg/kg）治疗组，每组n = 10。正常组和模型组小鼠均按0.1mL/10g（体重）的体积灌胃生理盐水（NS），阳性组腹腔注射环磷酰胺（CPA），剂量为20mg/kg（体重），GBEE治疗组分别按50、100、200mg/kg（体重）灌胃GBEE，每天1次，共20天。治疗后，计算肿瘤抑制率和抗转移率。通过免疫组织化学方法检测移植瘤中的微血管密度（MVD）。分别采用定量逆转录聚合酶链反应（qRT-PCR）或蛋白质免疫印迹法检测体外和体内血管内皮生长因子（VEGF）和VEGFR2 mRNA或Wnt3a、β-连环蛋白、VEGF、VEGFR2和p-Akt/Akt蛋白表达水平。
体外实验中，GBEE在5、10、20、40、80和160μg/mL剂量下呈剂量依赖性抑制LLC细胞生长。它能显著抑制LLC细胞中Wnt3a和β-连环蛋白蛋白表达以及VEGF和VEGFR2 mRNA含量。在体内，我们发现GBEE在50、100、200mg/kg剂量下呈剂量依赖性抑制LLC移植瘤生长，抑制肿瘤肺转移。CD34表达降低，这意味着MVD受到抑制，C57BL/6小鼠LLC移植瘤中β-连环蛋白、VEGF、VEGFR2和p-AKT/AKT蛋白表达以及VEGF和VEGFR2 mRNA表达水平也受到抑制。
GBEE通过抑制肿瘤血管生成发挥抗肿瘤和抗转移作用，这可能与其阻断LLC中Wnt/β-连环蛋白-VEGF信号通路的作用密切相关。
