替莫唑胺对人脑胶质瘤U87细胞增殖和血管生成的抑制作用

Inhibitory effect of temozolomide on proliferation and angiogenesis in human glioma U87 cells

目的:探讨替莫唑胺(Temozolomide,TMZ)对人胶质瘤U87细胞体内外生长及血管生成的影响.方法:利用流式细胞分析法测定TMZ诱导的U87细胞周期进程;利用EdU法检测TMZ对U87细胞增殖活性的影响;利用CCK-8法测定TMZ对U87细胞存活性的作用;利用裸鼠成瘤实验检测TMZ在体内对U87异体移植瘤生长的作用;免疫组织化学法检测肿瘤中Ki-67,BDNF及CD31表达,利用鸡胚绒毛尿囊膜(chicken chorioallantoic membrane,CAM)模型检测TMZ对体内血管生成的影响;ELISA实验检测TMZ对U87细胞中血管生成相关蛋白VEGF,TGF-β和FGF的分泌水平的影响.结果:TMZ增加了U87细胞中G0/G1,G2期细胞的比例,降低了S期细胞的比例,抑制了U87细胞在体外的存活和增殖.TMZ治疗抑制了裸鼠移植瘤的增长及肿瘤中血管内皮标志物的表达,降低了U87细胞中血管生成相关蛋白的水平.结论:TMZ在体内外抑制了胶质瘤U87细胞增殖和血管的生成,这种抑制作用可能与降低Ki-67,BDNF,VEGF,TGF-β和FGF蛋白表达相关.

Objective:To investigate the effects of Temozolomide (TMZ) on the growth and angiogenesis of human glioma U87 cells in vitro and in vivo. Methods:Flow cytometry was used to determine the cell cycle progression of U87 cells induced by TMZ, EdU method was used to detect the effect of TMZ on the proliferation activity of U87 cells, CCK-8 method was used to determine the effect of TMZ on the survival of U87 cells, nude mice tumorigenesis experiment was used to detect the effect of TMZ on the growth of U87 xenografts in vivo, and immunohistochemical method was used to detect the expressions of Ki-67, BDNF and CD31 in tumors. Angiogenesis was detected by chicken chorioallantoic membrane (CAM) model. The effect of TMZ on the secretion of angiogenesis-related proteins VEGF, TGF-βand FGF in U87 cells were detected by ELISA. Results:TMZ increased the proportion of G0/G1 and G2 phase cells in U87 cells, decreased the proportion of S phase cells and inhibited the survival and proliferation of U87 cells in vitro. TMZ therapy inhibited the growth of transplanted tumors and the expression of vascular endothelial markers in nude mice, and rEdUced the level of angiogenesis-related proteins in U87 cells. Conclusion:TMZ inhibits the angiogenesis of glioma U87 cells in vitro and in vivo. This inhibitory effect may be associated with decreased the protein expressions of Ki-67, BDNF, VEGF, TGF-β and FGF.