人参皂苷Rg3抗骨肉瘤LJH-OS细胞裸鼠移植瘤的作用及其对肿瘤血管生成拟态的影响

Effect of ginsenoside-Rg3 on osteosrcoma cells LJH-OS xenograft in nude mice and vasculogenic mimicry

目的 观察人参皂苷Rg3抗骨肉瘤LJH-OS细胞裸鼠移植瘤的作用及对肿瘤血管生成拟态（VM）的影响.方法 建立骨肉瘤裸鼠胫骨移植瘤动物模型,采用不同浓度（0、1、10 mg/L）Rg3瘤内注射给药,观察移植瘤体积和治疗变化;采用免疫组织化学双重染色法观察骨肉瘤VM密度;免疫组织化学和Western blot分别检测移植瘤中缺氧诱导因子-α（HIF-α）及基质金属蛋白酶-2（MMP-2）表达水平.体外建立三维培养系统,观察不同浓度（0、1、10 mg/L） Rg3对骨肉瘤LJH-OS细胞VM的影响.结果 人参皂苷Rg3可破坏骨肉瘤细胞体外培养的VM结构.通过瘤内注射Rg3,可显著降低体内骨肉瘤瘤体体积、质量、VM密度、HIF-α及MMP-2表达水平,并呈现剂量依赖性.当Rg3浓度分别为0、1、10 mg/L时,瘤体积（cm3）、瘤质量（g）、VM密度分别为：1.56 ±0.78、1.93±0.82、7.56±1.01;0.84±0.23、1.19 ±0.51、6.34±1.17;0.34±0.19、0.58 ±0.32、3.23±0.78;HIF-1α、MMP-2的表达水平分别为：（0.58±0.11）％、（0.47±0.09）％;（0.42±0.07）％、（0.33±0.05）％;（0.24±0.06）％、（0.21±0.07）.结论 人参皂苷Rg3可显著抑制骨肉瘤LJH-OS细胞裸鼠移植瘤的生长,下调HIF-α、MMP-2表达水平并破坏骨肉瘤VM是其可能作用机制.

Objective To evaluate the effect of ginsenoside-Rg3 on osteosrcoma cells LJH-OS xenograft in nude mice and vasculogenic mimicry （VM）.Methods Xenotransplanted models of nude mice were established with osteosarcoma cell LJH-OS.Different concentrations of Rg3 （0,1,10 mg/L）were injected inside tumor and the changes of volume and weight in transplanted tumors were observed.Immunohistochemistry double staining was applied to observe osteosarcoma VM density.Immunohistochemistry and Western blotting were used to detect the expression level of hypoxia induced factor-α （HIF-α） and matrix metalloproteinase-2 （MMP-2）.A three-dimensional cells culture system was established.The inhibitory effect of different concentration Rg3 （0,1,10 mg/L） on VM of LJH-OS cells was observed in vitro.Results In vitro,ginsenoside-Rg3 can be damaged VM of osteosarcoma cells.By tumor injection of Rg3,can significantly reduce the volume,quality,VM density,HIF-α and MMP-2 expression level of osteosarcoma tumors,and show dose dependent.Different concentration Rg3 （0、1、10 mg/L）,Volume （cm3）,weight（g）,VM density in nude mice：1.56 ±0.78,1.93 ±0.82,7.56 ± 1.01;0.84 ±0.23,1.19 ±0.51,6.34 ± 1.17;0.34 ±0.19,0.58 ±0.32,3.23 ±0.78;HIF-1α,MMP-2 in nude mice：（0.58±0.11）％,（0.47±0.09）％;（0.42±0.07）％,（0.33±0.05）;（0.24±0.06）,（0.21 ± 0.07）.Conclusion Ginsenoside-Rg3 can significantly inhibit the growth of osteosarcoma LJH-OS xenografts in nude mice.The mechanism may be due to down-regulation the expression of HIF-α and MMP-2 and inhibit VM.