天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (7): 745-748.doi: 10.11958/j.issn.0253-9896.2015.07.012

• 实验研究 • 上一篇    下一篇

PI-88 对 TE-13 细胞及裸鼠移植瘤中乙酰肝素酶表达的影响

朱辉，何明，陈新，李宝重，徐新建，李飞   

  1. 河北医科大学第四医院胸外科 （邮编 050011）
  • 收稿日期:2015-01-21 修回日期:2015-03-24 出版日期:2015-07-15 发布日期:2015-07-15
  • 通讯作者: 朱辉 E-mail:zhdzuj@sina.com E-mail:zhdzuj@sina.com
  • 作者简介:朱辉 （1977）， 男， 副教授， 博士， 主要从事胸部恶性肿瘤基础与临床研究
  • 基金资助:
    河北省卫生厅重点科研基金项目 （20120130）

The influence of PI-88 on heparanase protein expression of human esophageal cancer cell and xenograft of nude mice

ZHU Hui, HE Ming, CHEN Xin, LI Baozhong, XU Xinjian, LI Fei   

  1. Department of Thoracic Surgery, Fourth Hospital of Hebei Medical University，Shijiazhuang 050011, China
  • Received:2015-01-21 Revised:2015-03-24 Published:2015-07-15 Online:2015-07-15
  • Contact: ZHU Hui E-mail:zhdzuj@sina.com E-mail:zhdzuj@sina.com

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摘要： 目的 探讨硫酸化寡聚多糖 （PI-88） 对食管鳞癌细胞株 TE-13 乙酰肝素酶 （Hpa） 表达的影响， 以及对裸鼠人食管癌移植瘤生长、 血管生成及 Hpa 蛋白表达的影响。方法 体外培养 TE-13 细胞， 实验分为 A 组（对照组）、 B组 （15 mg/L PI-88 干预组） 和 C 组 （30 mg/L PI-88 干预组）。培养 36 h 后 Western blot 检测各组细胞 Hpa 蛋白表达。选取 10 只 BALB/c/nu 裸鼠， 皮下接种 TE-13 细胞建立食管癌裸鼠移植瘤模型， 建模成功后随机均分为观察组及对照组。观察组腹腔注射 PI-88 40 mg/ （kg·d）， 对照组给予等剂量生理盐水， 连续注射 14 d。每 2 d 测量移植瘤体积，注射第 14 天处死动物， 剥除移植瘤行 CD34 染色， 检测微血管密度（MVD）。Western blot 及免疫组化染色检测 Hpa蛋白表达。结果 与 A 组相比， B、 C 组细胞 Hpa 蛋白表达量明显降低 （P < 0.001）， 且存在药物浓度依赖性。观察组裸鼠移植瘤体积、 MVD 值、 Hpa 蛋白表达水平及阳性细胞数均低于对照组（均 P < 0.05）。结论 PI-88 可抑制 TE-13 细胞及食管癌裸鼠皮下移植瘤中 Hpa 表达， 并抑制肿瘤生长和血管生成。

关键词: 食管肿瘤, 硫酸化寡聚多糖, TE-13 细胞, 乙酰肝素酶, 印迹法, 蛋白质, 裸鼠, 移植瘤, 微血管密度, 免疫组织化学

Abstract: Objective To explore the inhibitory effects of sulfated oligosaccharides PI- 88 on the heparanase protein expression of human esophageal squamous cancer cell (ESCC) line TE-13, and to explore the effects of growth, angiogenesis and heparanase protein expression on ESCC xenografts of nude mice. Methods TE-13 cells were cultured and divided into three groups: group A (control group), group B (15 mg/L PI-88) and group C (30 mg/L PI-88). Heparanase protein expression of TE-
13 cells was measured by Western blot assay after being cultured for 36 h. The ESCC suspension was injected subcutaneously in 10 BALB/c/nu mice to build up ESCC xenograft model. The model mice were divided randomly into observation group and control group (5 mice per group). The mice in observation group received 40 mg/(kg·d) PI-88. The mice in control group only received the same volume of saline at the same time. Both PI-88 and saline were daily administrated for 14 days. Every 2 days, the volume of xeongrafts were measured and the mice were executed at the 14th day. CD34 immunohistochemical staining was used to detect the micro vessel density (MVD) of xenografts. Western blot assay and immunohistochemical staining were used to detect the heparanase protein expression of xenografts. Results The heparanase protein expressions of TE-13 cells were significantly decreased in group B and group C than those of group A (P < 0.001), with a kind of PI-88 dose-dependent manner. The volume, MVD and heparanase protein expression of xenografts were significantly lower in observation group than those of control group (P < 0.05). Conclusion The heparanase protein expression in TE-13 cells can be inhibited by PI-88 in vitro and vivo. Furthermore, the growth and angiogenesis of ESCC xenografts were also inhibited by PI-88.

Key words: esophageal neoplasms, PI- 88, TE- 13 cell, heparanase, blotting, western, mice, nude, heterografts, microvessel density, immunohistochemistry