摘要
目的:观察内皮抑素(ES)对裸鼠骨肉瘤皮下移植模型血管生成和肿瘤生长的抑制作用及其机制。方法:Transwell双室模型观察ES对骨肉瘤细胞9901诱导内皮细胞迁徙的影响;随机将30只骨肉瘤荷瘤裸鼠分为对照组、ES低剂量组[0.75mg(kg·d)-1]和高剂量组[1.5mg(kg·d)-1],瘤体及瘤体周围注射ES,1次/d,连续给药14d,观察肿瘤生长情况,计算抑瘤率。肿瘤组织进行病理学观察及免疫组化染色,计数坏死组织百分比,微血管密度以及细胞增殖和凋亡指数;动物处死前3个小时尾静脉注射EF5,激光共聚焦显微镜观察组织缺氧及血管标记。结果:ES低剂量组(350ng·ml-1)和高剂量组(700ng·ml-1)对内皮细胞迁徙有明显抑制作用,且高剂量组明显高于低剂量组(P<0.01);高剂量组和低剂量组平均瘤重明显小于对照组,抑瘤率分别为35.83%和22.92%(P<0.01);正常对照组、低剂量组、高剂量组的坏死百分比分别为2.40%、4.68%和9.52%(P<0.01),细胞增殖指数(PI)分别为66.44%、46.11%和25.11%(P<0.01),细胞凋亡指数(AI)分别为2.69%、7.58%和11.53%(P<0.01)。激光共聚焦观察ES治疗组绿色荧光量减少,血管稀疏;肿瘤细胞缺氧增加,呈弥漫的强红色。结论:ES可显著地抑制骨肉瘤细胞诱导的内皮细胞迁徙,以及人骨肉瘤的生长和发展,且呈剂量依赖效应,其可能机制为抑制血管生成,增加肿瘤细胞缺氧和凋亡。
Objective： To evaluate the inhibitory action of endostatin （ES） on human osteosarcoma tumor growth in nude mice and its possible mechanism. Methods： The inhibitory role of ES on endothelial cell migration induced by osteosarcoma cell line 9901 was observed using a transwell model. Thirty Balb/c nu/nu mice implanted with osteosarcoma were randomly divided into the control, ES low dose （0.75 mg/Kg.d） and ES high dose （1.5 mg/Kg.d） groups. ES was injected intratumorally and around the tumor, once a day for 14 days. The tumor volumes were measured and the tumor inhibition rate （TIR） was calculated, The percentages of necrotic tissue, microvessel density, proliferation index （PI） and apoptotic index （AI） were detected by pathohistologic observation and immunohistochemical staining. The mice were sacrificed 3 h after EF5 injection via tail vein, and then the hypoxia of tumor cells and microvessels was observed using laser confocal microscopy. Results： Both the low dose （350 ng/ml） and high dose （700 ng/ml） ES groups showed a marked inhibition of endothelial cell migration induced by osteosarcoma cells, while the high dose showed much stronger inhibition than the low dose （P〈0.01）. The tumor volume was significantly different among the 3 groups. The TIR in the high dose and the low dose groups were 35.8% and 22.9%, respectively. In the control group, the ES low dose group and the ES high dose group, the percentage of necrotic tissue was 2.40%, 4.68% and 9.52%, the PI was 66.44%, 46.11% and 25.11%, and the AI was 2.69%, 7.58% and 11.53%, respectively （P〈0.01）. Much less green fluorescence （microvessel） and stronger red fluorescence （hypoxia） were apparent in the tumors treated with ES when observed with laser confocal microscopy. Conclusions： The results show that ES markedly inhibits endothelial cell migration in vitro and significantly restrains the growth and development of osteosarcoma in vivo. The mechanism may be due to anti-angiogenesis properties associa
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2007年第1期52-55,共4页
Chinese Journal of Clinical Oncology
基金
国家自然科学基金资助(编号:30340044
30472004)
