天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (9): 1069-1072.doi: 10.11958/20160064

• 细胞与分子生物学 • 上一篇    下一篇

人参皂苷 Rg3 对人乳腺癌 MCF-7 细胞血管生成拟态的影响

黎国栋 1， 杨慧 2， 陆颖 2△   

  1. 1广西医科大学第四附属医院/柳州市工人医院药学部 （邮编 545055）， 2肿瘤科
  • 收稿日期:2016-02-22 修回日期:2016-04-25 出版日期:2016-09-15 发布日期:2016-09-28
  • 基金资助:

    广西壮族自治区卫生厅计划课题 （Z2014394）

Effects of ginsenoside Rg3 on vasculogenic mimicry of breast cancer MCF-7 cell line

LI Guodong1， YANG Hui2， LU Ying2△   

  1. 1 Department of Pharmacy, 2 Department of Medical Oncology, the Fourth Affiliated Hospital of Guangxi Medical University / Liuzhou Worker’ s Hospital, Liuzhou 545055, China
  • Received:2016-02-22 Revised:2016-04-25 Published:2016-09-15 Online:2016-09-28

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摘要： 摘要： 目的 探讨人参皂苷 Rg3 对人乳腺癌 MCF-7 细胞体外血管生成拟态的抑制作用及其机制。方法 取 MCF-7 对数生长期细胞分为实验组和对照组， 实验组加入含有终浓度为 10、 20、 50、 100、 150 及 300 mg/L 的 Rg3 完 全培养基； 对照组加入含 DMSO 完全培养基。CCK-8 法分析 Rg3 对 MCF-7 细胞增殖的影响， 分析实验组各浓度下 的抑制率和 Rg3 半数抑制浓度 （IC50） 并确定实验浓度。体外管道形成抑制实验观察 Rg3 对 MCF-7 细胞血管生成拟 态的影响。实时荧光定量 PCR 法检测 Rg3 对细胞血管内皮生长因子-A（VEGF-A）和基质金属蛋白酶（MMP） 9 mRNA 的表达水平的影响。蛋白免疫印迹法检测 Rg3 对细胞低氧诱导因子 （HIF） -1α、 VEGF-A 和 MMP9 蛋白表达 的影响。双抗夹心酶联免疫法测定 Rg3 对细胞培养上清液中 VEGF-A 、 MMP9 蛋白表达的影响。结果 人参皂苷 Rg3 可以有效抑制 MCF-7 细胞增殖， 其平均 IC50为 （115.34±8.50） mg/L； 选取 50、 100 和 150 mg/L 的 Rg3 作为实验浓 度。50、 100 和 150 mg/L 组形成的管状结构数目分别为 （19.0±1.0）、（15.0±1.5）、（10.0±1.7） 个/视野， 呈逐渐降低趋势； 且均较对照组（22.0±1.8）减少（均 P < 0.05）。对照组、 50 mg/L 组、 100 mg/L 组及 150 mg/L 组细胞中的 VEGF-A 和 MMP9 的 mRNA 和蛋白表达水平、 HIF-1α 的蛋白表达水平均呈逐渐下降趋势 （P < 0.05）。对照组、 50 mg/L 组、 100 mg/ L 组及 150 mg/L 组细胞培养上清液中的 VEGF-A 蛋白呈逐渐下降趋势 （P < 0.05）； 后 3 组的 MMP9 蛋白表达水平均呈 逐渐下降趋势 （P < 0.05）， 但 50 mg/L 组与对照组的 MMP9 蛋白表达水平差异无统计学意义。结论 人参皂苷 Rg3 可以抑制 MCF-7细胞体外血管形成拟态的能力， 其分子机制可能与抑制 MCF-7细胞中 HIF-1α、 VEGF-A和 MMP9的蛋白表达有关

关键词: 乳腺肿瘤, 血管内皮生长因子 A, 基质金属蛋白酶 9, 低氧诱导因子-1α, MCF-7 细胞, 血管生成拟态, 人 参皂苷 Rg3

Abstract: Abstract: Objective To investigate the inhibitory effects and mechanism of ginsenoside Rg3 on vasculogenic mimicry of human breast cancer MCF-7 cell line. Methods The MCF-7 cells at logarithmic growth phase were obtained, and were cultured with different concentrations (0, 20, 50, 100, 150 and 300 mg/L) of ginsenoside Rg3. Cells cultured without Rg3 were served as controls. The IC50 were determined by CCK8 assay and anti-angiogenic effects were performed for testing the potential of tube-like structure (TLSs) formation. The expression levels of VEGF-A, MMP9 and HIF-1α were detected by Western blotting and real-time PCR. The secreted contents of VEGF-A and MMP9 were detected by enzyme linked immunosorbent assay (ELISA). Results The ginsenoside Rg3 suppressed the proliferation of MCF-7 cells in a dosedependent manner, in which IC50 was (115.34±8.50) mg/L. The formation numbers of TLSs in MCF-7 cells were significantly inhibited by Rg3 in concentration dependent manner in 50 mg/L, 100 mg/L and 150 mg/L for (19.0±1.0), (15.0±1.5), and (10.0±1.7) vs. controls (22.0±1.8, F=150.805, P < 0.05). The mRNA and protein expression levels of VEGF-A, MMP9 and HIF-1α protein were inhibited by 50 mg/L,100 mg/L and 150 mg/L Rg3 vs. controls (P < 0.05). Meanwhile, the contents of VEGF-A in MCF-7 cell supernatant was down-regulated by 50 mg/L,100 mg/L and 150 mg/L Rg3 vs. controls (P < 0.05). The contents of MMP-9 in MCF-7 cell supernatant was down-regulated by 100 mg/L and 150 mg/L Rg3 vs. controls (P < 0.05). There was no significant difference in MMP- 9 expression between 50 mg/L group and control group. Conclusion The ginsenoside Rg3 is able to inhibit the vasculogenic mimicry of MCF- 7 cells, which may be related with the down-regulation of VEGF-A, MMP9 and HIF-1α.

Key words:  breast neoplasms, vascular endothelial growth factor A, matrix metalloproteinase 9, hypoxia inducible factor-1α, michigan cancer foundation-7 cell, vasuclogenic mimicry, ginsenoside Rg3