摘要
目的探讨骨肉瘤细胞的仿血管发生现象，骨肉瘤细胞是否具有血管形成相关基因的表达。方法应用Ⅰ型胶原蛋白凝胶和Matrigel的三维培养基对骨肉瘤细胞进行培养，并运用电镜、光镜观察这些类血管样结构的形态学构成；应用逆转录．聚合酶链反应（RT-PCR）技术检测骨肉瘤细胞血管形成相关基因（VEGF、Flt-1、KDR、Ang-1、Tie-2、Ang-2、Tie-1、EPHA2、CD31、Thrombin、VE-Cadherin）的表达。结果电镜、光镜下观察到骨肉瘤细胞在I型胶原蛋白凝胶和Matrigel的三维培养基中能够形成良好的类血管样的管道结构；骨肉瘤细胞类内皮细胞和内皮前驱细胞，具有血管形成相关基因（VEGF、Flt-1、KDR、Ang-1、Tie-2、EPHA2、CD31、Thrombin、VE-Cadherin）的表达。结论骨肉瘤细胞具有仿血管发生的能力和血管形成相关基因的表达，三维培养是研究肿瘤细胞仿血管发生现象较好的工具。
Objective To analyze vasculogenic mimicry in osteosarcoma ceUs （MG63） and investigate the function of the gene correlated with the internal mechanism of the vasculogenic mimicry. Methods The structures of the tubular networks were observed in three-dimensional culture containing type Ⅰ collagen or the Matrigei with HE stain under a phase contrast microscope and electron microscope. The genes （VEGF,Fh-1 ,KDR,Ang-1 ,Tie-2,Ang-2,Tie-1 ,EPHA2,CD31 ,Thrombin,VE-Cadherin） associated with the origin of the blood vessels were detected in osteosarcoma cell by using.RT-PCR. Results Under the ligt microscopy and electron microscopy, it was found that high aggressive osteosarcoma cells （MG63） could form networks containing channels when grown in three-dimensional culture in the absence of endothelial cells. The expression of the genes （ VEGF, Fit-1, KDR, Ang-1, Tie-2, EPHA2, CD31, Throm- bin,VE-Cadherin） associated with the origin of the blood vessels was detected in osteosarcoma cell （MC63）-like endothelial precursor cells. Conclusion Aggressive osteosarcoma cells may generate tumor cell-lined vascular channels and can express vascular cells-associated genes. The vascular cells-associated genes may be necessary for the formation of osteosarcoma-generated, vasculogenic-like, patterned networks and may be a target for therapy. In the process of comprehensive therapy,especially antiangiogenesis, the vasculogenie mimicry cann＇ t be ignored.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2008年第6期716-718,818,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30271314、3047170）
关键词
骨肉瘤
细胞培养
基因表达
Osteosarcoma
Cell culture
Gene expression
