天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (5): 453-456.doi: 10.11958/j.issn.0253-9896.2015.05.002

• 细胞与分子生物学 • 上一篇    下一篇

干扰素-γ对黑色素瘤细胞血管生成拟态的影响及机制

韩健 1， 孙保存 1△， 马跃美 2， 赵秀兰 1   

  1. 1天津医科大学病理学教研室 （邮编 300070）； 2天津医科大学外科手术学教研室
  • 收稿日期:2014-12-10 修回日期:2015-02-06 出版日期:2015-05-15 发布日期:2015-05-25
  • 通讯作者: 孙保存 E-mail： sunbaocun@aliyun.com E-mail:sunbaocun@aliyun.com
  • 作者简介:韩健 （1983）， 男， 助理实验师， 硕士在读， 主要从事肿瘤病理学研究
  • 基金资助:
    国家自然科学基金重点项目 （81230050）； 天津市科委重点项目 （10JCZDJC20400）

Effects and mechanism of interferon-gamma on vasculogenic mimicry of melanoma cells

HAN Jian1， SUN Baocun1△， MA Yuemei2， ZHAO Xiulan1   

  1. 1 Department of Pathology, Tianjin Medical University, Tianjin 300070, China; 2 Department of Surgery, Tianjin Medical University
  • Received:2014-12-10 Revised:2015-02-06 Published:2015-05-15 Online:2015-05-25
  • Contact: SUN Baocun,E-mail： sunbaocun@aliyun.com E-mail:sunbaocun@aliyun.com

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摘要： 摘要： 目的 研究干扰素-γ（IFN-γ）对人黑色素瘤细胞系 MUM-2B 迁移、 侵袭和血管生成拟态（VM）形成能力的影响。方法 将 MUM-2B 细胞体外培养后分为 3 组： 对照组在含 10%FBS 的 DMEM 培养基中培养； 实验 1 组另加入 10 μg/L IFN-γ； 实验 2 组另加入 100 μg/L IFN-γ。利用划痕实验和侵袭实验分析 IFN-γ对 MUM-2B 细胞迁移、 侵袭能力的影响， 三维培养观察 IFN-γ对 MUM-2B 细胞 VM 形成能力的影响， Western blot 检测 MUM-2B 细胞中血管内皮生长因子 （VEGF）表达的变化。结果 划痕实验示实验组细胞迁移距离低于对照组， 实验 2 组低于实验 1 组 （P < 0.05）； 侵袭实验示实验组细胞侵袭数量低于对照组， 实验 2 组低于实验 1 组 （P < 0.05）； 三维培养示对照组细胞能够形成 VM 管道结构， 实验组细胞不能形成明显的 VM 管道结构； Western blot 示实验组细胞中 VEGF 蛋白表达量低于对照组， 实验 2 组低于实验 1 组（P < 0.05）。结论 IFN-γ可以在体外抑制 MUM-2B 细胞的迁移和侵袭， 并通过抑制 VEGF 的表达而抑制其 VM 的形成。

关键词: 黑色素瘤, 干扰素γ, 重组, 血管内皮生长因子类, 肿瘤干细胞, 血管生成拟态

Abstract: Abstract： Objective To investigate the effects of interferon-gamma (IFN-γ) on migration， invasion and vasculogenic mimicry (VM) formation of human melanoma cell line MUM-2B. Methods MUM-2B cells were divided into three groups, control group (10%FBS in DMEM), treatment group1 (10 μg/L IFN-γ) and treatment group2 (100 μg/L IFN-γ). Different concentrations of IFN-γ were added in the culture medium of MUM-2B cells. Wound- healing assay and matrigel invasion assay were performed to examine the migration and invasion ability of MUM-2B cells. Three-D culture was used to observe the VM formation. The expression of vascular endothelial growth factor (VEGF) of MUM-2B cells was detected by Western blot assay. Results The result of wound-healing assay showed that the migration distance of cells was decreased in treat⁃ ment groups compared with that of control group. The migration distance of cells was decreased in treatment group2 com⁃ pared with that of treatment group 1(P < 0.05). The result of matrigel invasion assay showed that the number of invaded cells was decreased in treatment groups compared with that of control group, and which was significantly decreased in treatment group2 than that of treatment group1 (P < 0.05). The result of 3-D culture showed that cells in control group can form typical VM tube-like structures, whereas cells in treatment groups cannot. Western blot assay showed that the expression of VEGF protein was significantly decreased in treatment groups compared with that of control group, and the expression of VEGF pro⁃ tein was significantly decreased in treatment group2 than that of treatment group 1(P < 0.05). Conclusion These data sug⁃ gest that IFN-γ inhibits migration and invasion of MUM-2B cells, and inhibits VM formation by down regulating VEGF ex⁃ pression in vitro.

Key words: melanoma, interferon-gamma, recombinant, vascular endothelial growth factors, cancer stem cells, vasculogenic mimicry