国际肿瘤学杂志

国际肿瘤学杂志 ›› 2015, Vol. 42 ›› Issue (10): 721-725.doi: 10.3760/cma.j.issn.1673-422X.2015.10.001

• 论著 •    下一篇

索拉非尼抑制人肝癌裸鼠皮下移植瘤血管生成拟态作用研究

张月英,王兆朋,王朝霞,贾青,王恒孝,姜国胜,张维东   

  1. 250062 济南，山东省医学科学院基础医学研究所病理学与病理生理学研究室（张月英、王兆朋、王朝霞、贾青、王恒孝、张维东），免疫学研究室（姜国胜）
  • 收稿日期:2015-06-19 出版日期:2015-10-08 发布日期:2015-09-08
  • 通讯作者: 张维东 E-mail:zhangweidongkui@163.com
  • 基金资助:

    国家自然科学基金（81403150、81303077、81073102）

Sorafenib inhibits tumor growth through modulating vasculogenic mimicry in a hepatocellular carcinoma xenograft model

Wang Zhaopeng, Wang Zhaoxia, Jia Qing, Wang Hengxiao, Jiang Guosheng, Zhang Weidong   

  1. Department of Pathology and Pathophysiology, Institute of Basic Medicine, Shandong Academy of Medical Sciences, Ji′nan 250062, China
  • Received:2015-06-19 Online:2015-10-08 Published:2015-09-08
  • Contact: Zhang Weidong E-mail:zhangweidongkui@163.com

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被引次数

4

摘要： 目的 探讨索拉非尼对人肝癌裸鼠皮下移植瘤血管生成拟态的抑制作用及机制。方法 构建人肝癌细胞HepG2裸鼠皮下移植瘤模型，采用配对比较法将其随机分为荷瘤对照组和索拉非尼组（30 mg·kg-1·d-1）。绘制各组肿瘤生长曲线，应用免疫组织化学和过碘酸希夫反应双染法检测肿瘤血管生成拟态，同时利用免疫组织化学、Western blotting和实时荧光定量PCR检测HIF-1α、VEGFA、VEGFR1和MMP2基因表达。结果 与荷瘤对照组相比，索拉非尼组肿瘤体积明显减小［(809.69±208.71)mm3∶(1 678.00±313.29)mm3］，差异有统计学意义（t=6.103，P=0.030）。苏木精-伊红染色显示，对照组肿瘤组织瘤细胞生长旺盛，坏死范围小；索拉非尼治疗组肿瘤组织中有大片坏死区域。与对照组相比，索拉非尼组DiOC7标记的功能性血管数明显减少［（4.77±0.15）个∶（8.44±0.68）个］，差异有统计学意义（t=9.192，P=0.013）。索拉非尼组中血管生成拟态数量较荷瘤对照组明显减少［（1.04±0.46）个∶（2.66±0.42）个］，差异有统计学意义（t=4.510，P=0.041）。与荷瘤对照组比较，索拉非尼组CD31标记阳性血管数量明显减少［（1.26±0.14）个∶（3.42±0.10）个］，差异具有统计学意义（t=21.580，P=0.002）。进一步研究发现索拉非尼组HIF-1α（0.65±0.03∶1.00±0.00）、VEGFA（0.51±0.02∶1.00 ±0.00）、VEGFR-1（0.45±0.04∶1.00±0.00）和MMP-2（0.69±0.02∶1.00±0.00）在蛋白水平上表达明显下降（t=19.650，P=0.003；t=40.493，P=0.000；t=23.429，P=0.002；t=26.071，P=0.002）。同时也发现索拉非尼组HIF1α（0.78±0.05∶1.00±0.00）、VEGFA（0.52±0.05∶1.00±0.00）、VEGFR-1（0.45±0.02∶1.00±0.00）和 MMP-2（0.71±0.02∶1.00±0.00）在mRNA水平上表达也明显下降（t=6.840，P=0.021；t=27.710，P=0.001；t=62.740，P=0.000；t=23.850，P=0.002）。结论 索拉非尼抑制实验性肝癌生长和血管生成拟态可能与其抑制HIF-1α、VEGFA/VEGFR-1信号通路有关。

关键词: 肝肿瘤, 芳香烃受体核转位子, 血管生成抑制剂, 索拉非尼, 血管生成拟态

Abstract: Objective To investigate the effects and the mechanism of sorafenib on hepatocellular carcinoma growth and vasculogenic mimicry (VM) in mice. Methods A subcutaneous implantation mouse model of human hepatocellular carcinoma (HCC) HepG2 cells were established. Mice inoculated with HepG2 cells were randomly divided into the treatment group (sorafenib 30 mg·kg-1·d-1) and the control group by using of paired comparison method. Growth of established tumor xenografts was monitored at least twice weekly by vernier caliper measurements. VM was assessed by immunohistochemical assay and periodic acid schiff reaction (PAS) histochemical doublestaining. The expressions of HIF-1α, VEGFA, VEGFR-1 and MMP-2 in tumor tissues were also assessed by immunohistochemical assay, Western blotting and realtime quantitative PCR. Results The tumor volume in the sorafenib group was obviously decreased compared with the control group (809.69 mm3±208.71 mm3 vs 1 678.00 mm3±313.29 mm3), with a statistically significant difference (t=6.103, P=0.030). Haematoxylin and eosin (HE) staining showed that tumor tissues treated with sorafenib were characterized by obvious necrosis, but there were not the same cases in the control group. Sorafenib group significantly reduced the number of tumor functional vessel in HepG2 xenografts compared with the control group, as assessed by tumor vasculature uptake of DiOC7 (4.77±0.15 vs 8.44±0.68, t=9.192, P=0.013). The number of VM was significantly decreased by sorafenib (1.04±0.46 vs 2.66±0.42, t=4.510, P=0.041). Relative to controls, CD31positive vessels decreased after treatments (3.42±0.10 vs 1.26±0.14), with a statistically significant difference (t=21.580, P=0.002). Compared with the control group, the protein levels of HIF-1α (0.65±0.03 vs 1.00±0.00), VEGFA (0.51±0.02 vs 1.00 ±0.00), VEGFR-1 (0.45±0.04 vs 1.00±0.00) and MMP-2 (0.69±0.02 vs 1.00±0.00) were significantly decreased in the sorafenib group (t=19.650, P=0.003; t=40.493, P=0.000; t=23.429, P=0.002; t=26.071, P=0.002). Compared with the control group, the mRNA levels of HIF-1α (0.78±0.05 vs 1.00±0.00), VEGFA (0.52±0.05 vs 1.00±0.00), VEGFR-1 (0.45±0.02 vs 1.00±0.00) and MMP-2 (0.71±0.02 vs 1.00±0.00) were also significantly decreased in sorafenib group (t=6.840, P=0.021; t=27.710, P=0.001; t=62.740, P=0.000; t=23.850, P=0.002). Conclusion Sorafenib can inhibit the tumor growth and VM channels formation, which may be related with the HIF1α and VEGFA/VEGFR-1 signaling pathway.

Key words: Liver neoplasms, Aryl hydrocarbon receptor nuclear translocator, Angiogenesis inhibitors, Sorafenib, Vasculogenic mimicry