Joint Study on Angiogenesis and Growth Factor Responses

M.Kuwano， M.Ono， Y.Sato和K.Kohno与K.Miyazono， D.Rifkin和h . f.合作完成了血管生成和生长因子反应的联合研究。我们得到的结果如下所述。我们最近建立了体外血管生成模型，将I型胶原凝胶上的血管内皮细胞内腔与外腔中其他类型的细胞共培养。体外培养人软骨细胞可抑制EGF/TGFalpha依赖性血管内皮细胞的管状形成。这种由软骨细胞诱导的管形成抑制被抗TGF β抗体部分消除，提示TGF β部分参与了人软骨细胞依赖性血管的形成。H-ras癌基因的引入降低了小鼠Balb/3T3细胞中EGF与所有表面EGF受体的结合活性。在H-ras转染中，EGF受体基因表达被破坏，但V-myc的共同引入使EGF受体恢复到正常水平。引入编码转录失活产物的显性阴性c-jun突变体，抑制了H-ras依赖性AP-1 （Jun/Fos）的诱导，并恢复了H-ras癌基因对EGF结合的下调。在包括炎症在内的各种病理条件下都会诱导氧自由基。过氧化氢（H_2O_2）诱导人微血管内皮细胞形成管状结构，同时给予NFkB反义序列抑制H_2O_2依赖性管状结构的形成，在抗il - 8抗体存在时几乎完全消除。氧化刺激后血管内皮细胞的管状形态形成似乎与NFkB和IL8.4有关。临床上使用的依格列定在10^< -6> ~ 10^<-4>M范围内作为抗胃溃疡药物，特异性地抑制了血管内皮细胞的细胞增殖和管状形成。口服伊索格拉定30 ~ 120mg /kg/天抑制人肿瘤细胞在小鼠体内的生长和血管生成。少

M.Kuwano, M.Ono, Y.Sato and K.Kohno have done the joint study on angiogenesis and growth factor responses in collaboration with K.Miyazono, D.Rifkin and H-f.Kung, and we obtained results as described below.1. We have recently established angiogenesis model in vitro in wihch vascular endothelial cells on type I collagen gel in an inner chamber are co-cultured with other types of cells in an outer chamber. EGF/TGFalpha dependent tube formation of vascular endothelial cells was inhibited when human chondrocytes were co-cultured in the outer chamger. This chondrocype-induced inhibition of tube fomation was partly abrogated by administraion of anti-TGF beta antibody, suggesting that TGF beta is partly involved in the human chondrocyte-dependent avascularily.2. Introduction of H-ras oncogene decreased the EGF binding activity to all surface EGF receptor in mouse Balb/3T3 cells. EGF receptor gene expression was abrogated in the H-ras transfectants, but co-introduction of V-myc restored the EG … More F receptor to normal levels.Introduction of dominant-negative c-jun mutant encoding a transcriptionally inactive product inhibited the H-ras dependent AP-1 (Jun/Fos) induction, and also restored down-regulation of EGF binding by H-ras oncogene.3. Oxygen radicals are induced under various pathological conditions including inflamation. Treatment with hydrogen peroxide (H_2O_2) of human microvascular endothelial cells induced development of tube-like structres, and co-administration of antisense sequence of NFkB inhibited the H_2O_2 dependent tubular morphogenesis was almost completely abrogated when anti-IL8 antibody was present. Tubular morphogenesis of vascular endothelial cells after the oxidative stimuli appeared to be associated with NFkB and IL8.4. Irogladine used clinically as an anti-gasTriculcer agent at 10 ^<-6> to 10^<-4>M inhibited specifically cell proliferation and tube formation of vascular endothelial cells. Oral administration of irsogladine at 30 to 120 mg/kg/day inhibited both tumor growth and tumor angiogenesis when human tumor cells were transplanted in mice. Less