去甲二氢愈创木酸对胶质瘤诱导的血管内皮细胞成型的作用

THE EFFECT OF NDGA ON GLIOMA-INDUCED ENDOTHELIAL TUBULAR MORPHOGENESIS AND ITS MECHANISM

目的 观察去甲二氢愈创木酸 (NDGA)对胶质瘤细胞诱导的内皮细胞成型的影响并初步探讨其机理。 方法 采用在 型胶原凝胶上的细胞立体培养和内皮 -胶质瘤细胞的联合培养法 ,观测人恶性胶质瘤细胞系SHG- 44细胞或其条件培养基对人脐静脉内皮细胞系 ECV- 30 4细胞形成管腔样结构的影响 ,同时以免疫组织化学结合图像分析的方法检测 SHG- 44细胞 VEGF、b FGF蛋白的表达。 结果 10 0 μmol/ L 的 NDGA作用 1～ 3d后 ,SHG- 44细胞 VEGF、b FGF的表达明显降低。联合培养 3d左右 ,胶质瘤 SHG- 44细胞及其条件培养基均能诱导内皮细胞在 型胶原凝胶上拉长并形成管腔样结构 ;而 10 0 μm ol/ L 的 NDGA可明显抑制内皮细胞变长和管腔样结构的形成 (P<0 .0 1)。 结论 NDGA对胶质瘤细胞诱导的血管成型有抑制作用 ,此作用可能与瘤细胞 VEGF、b FGF表达减低有关 ,提示

Objective\ To observe the effect of NDGA on glioma\|induced tubular morphogenesis of endothelial cells, and its mechanism. Methods\ We applied the glioma\|endothelial cell coculture and collegen I gel\|based 3D culture to investigate the effect of NDGA on the formation of tubular structures of human umbilical vein endothelial cells(HUVEC)cell line ECV\|304 cell, which was induced by human malignant glioma cell line SHG\|44 or its conditioned media.Meanwhile the expression levels of VEGF and bFGF in SHG\|44 cells were investigated using the techniques of immunohistochemistry and image analysis. Results\ The Human malignant glioma cell line SHG\|44 after treated with 100μmol/L NDGA for 1 to 3 days showed that the expression of VEGF and bFGF was declined at protein levels. Either malignant glioma cell line SHG\|44 or its conditioned media could induce ECV\|304 cells to lengthen and form tubular structure on the gel of type I collegen. After addition of 100 μmol/L NDGA the length of endothelial cell and quantity of tubular structure, index of the glioma\|induced morphogenesis of endothelial cells were significantly declined. Conclusion\ The above results suggest that NDGA suppresses the tubular morphogenesis and that the decreased expression of VEGF and bFGF may be the molecular mechanism of anti\|angiogenesis effect of NDGA.\;