干扰素-γ对黑色素瘤细胞血管生成拟态的影响及机制

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

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10μg/L IFN-γ;实验2组另加入100μg/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。

Objective To investigate the effects of interferon-gamma（IFN-γ） on migration,invasion and vasculogenicmimicry（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）.Differentconcentrations of IFN-γ were added in the culture medium of MUM-2B cells.Wound-healing assay and matrigel invasionassay were performed to examine the migration and invasion ability of MUM-2B cells.Three-D culture was used to observethe VM formation.The expression of vascular endothelial growth factor（VEGF） of MUM-2B cells was detected by Westernblot 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 cellswas decreased in treatment groups compared with that of control group,and which was significantly decreased in treatmentgroup2 than that of treatment group1（P〈0.05）.The result of 3-D culture showed that cells in control group can form typicalVM tube-like structures,whereas cells in treatment groups cannot.Western blot assay showed that the expression of VEGFprotein 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.