Cui Guo-Hui, Chen Wei-Hua, Xue Ke-Ying, Liu Fang, Chen Yan
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2006 Oct;14(5):1008-12.
In order to study the relation of antitumour mechanisms of triptolide with neovascularization, the effect of triptolide and tumour necrosis factor (TNF)-alpha on the expression of vascular endothelial growth factor (VEGF) in Raji cell lines and their effect on angiogenesis in human umbilical vein endothelial cells (HUVECs)-derived cell line ECV304 were investigated. The inhibitory rate of cells treated by triptolide detected by MTT; the ELISA was employed to study the VEGF content secreted by Raji cell lines; angiogenesis was tested by network formation of endothelial cells on Matrigel, and the mRNA level of VEGF was measured by RT-PCR. The results showed that treatment of Raji cells with triptolide resulted in significantly enhanced antiproliferative effects in dose- and time-dependent manner. The content of VEGF secreted by Raji cells was increased by TNF-alpha and was suppressed by triptolide (P < 0.01). The mRNA expressions of VEGF(165) and VEGF(121) (containing 165 and 121 amino acid residues, respectively) could be detected in all fractions. TNF-alpha augmented the expression of VEGF(165) and VEGF(121) mRNA when triptolide reduced the expression (P < 0.01). No network and cord were formed in control and triptolide group. There was tube formation on matrigel in the supernatants of Raji culture group and the supernatants groups treated by VEGF and TNF-alpha in Raji cell. It is concluded that the expressions of VEGF in Raji cells are increased by TNF-alpha and suppressed by triptolide. VEGF and TNF-alpha induce angiogenesis and triptolide inhibits angiogenesis in ECV304 cells.
为研究雷公藤甲素抗肿瘤机制与新血管形成的关系，探讨雷公藤甲素及肿瘤坏死因子-α（TNF-α）对Raji细胞系血管内皮生长因子（VEGF）表达的影响及其对人脐静脉内皮细胞（HUVECs）来源的细胞系ECV304血管生成的作用。采用MTT法检测雷公藤甲素处理细胞的抑制率；用ELISA法研究Raji细胞系分泌的VEGF含量；通过基质胶上内皮细胞形成网络来检测血管生成，并用RT-PCR法检测VEGF的mRNA水平。结果显示，雷公藤甲素处理Raji细胞后，其抗增殖作用呈剂量和时间依赖性显著增强。Raji细胞分泌的VEGF含量在TNF-α作用下升高，而在雷公藤甲素作用下受到抑制（P<0.01）。在所有组分中均可检测到VEGF(165)和VEGF(121)（分别含165和121个氨基酸残基）的mRNA表达。当雷公藤甲素降低VEGF(165)和VEGF(121) mRNA表达时，TNF-α增强其表达（P<0.01）。对照组和雷公藤甲素组未形成网络和条索。Raji培养组上清以及Raji细胞经VEGF和TNF-α处理后的上清组在基质胶上有管腔形成。结论：TNF-α增加Raji细胞中VEGF的表达，雷公藤甲素抑制其表达。VEGF和TNF-α诱导血管生成，而雷公藤甲素抑制ECV304细胞的血管生成。
