血管抑素抑制胶质瘤的实验研究  

Inhibition of Gliomas by Gene Transfer of Angiostatin in Nude Mice

作　　者：刘猛[1] 孙学英[2] 刘玉光[1] 王宏伟[1] 张良文[1] 吴承远[1] 

机构地区：[1]山东大学齐鲁医院神经外科,山东济南250012 [2]山东大学齐鲁医院普外科,山东济南250012

出　　处：《中国神经肿瘤杂志》2006年第2期129-133,共5页Chinese Journal of Neuro-Oncology

摘　　要：背景与目的:近年来血管抑素在胶质瘤治疗中的意义倍受关注。本研究探讨血管抑素重组表达质粒对裸鼠皮下胶质瘤血管生成、细胞凋亡和肿瘤生长的影响,以寻求治疗胶质瘤的有效方法。方法:取Balb/c裸小鼠22只,建立裸鼠皮下胶质瘤模型,6～9天左右皮下肿瘤平均直径达到4mm,随机分为对照组和As治疗组,每组11只,分别注入100μl纯化pcDNA3和As重组质粒,注射2天后各组随机取5只处死取肿瘤作检测。分别做HE染色,As蛋白免疫组化染色,并行血管内皮细胞标记物CD31检测,观察肿瘤血管化的情况,并行TUNEL法检测肿瘤细胞的凋亡。剩余每组6只每隔3天测量肿瘤长、短径,绘制肿瘤生长曲线图。结果:As基因治疗可抑制肿瘤的生长,对照组基因注射后15天肿瘤体积为(1519.88±256.54)mm3,As治疗组肿瘤体积为(1148.03±191.51)mm3,两者差异有显著性(P<0.05)。As治疗组与对照组相比血管密度明显减小。两组凋亡指数差异亦有显著性(P<0.05),凋亡细胞集中在肿瘤坏死区周围,细胞核固缩呈圆形或椭圆形团块,典型者染色质呈新月形或蚕豆状浓染。结论:As具有抑制血管生成的作用,并可诱导肿瘤细胞的凋亡,对裸鼠皮下胶质瘤的生长具有延缓作用。BACKGROUND ＆ OBJECTIVE： Recently, angiostatin has been intensively investigated in treatment of glioma. This investigation was to explore the inhibitory effects of angiostatin （As） on angiogenesis, tumor apoptosis and tumor growth of subcutaneous gliomas in nude mice. METHODS： C6 cells （1×10^6/100 μl） were subcutaneously implanted into 4- to 6-week-old BALb/c nude mice to establish glioma model. When tumors reached 0.4 cm （30-35mm^3） in diameter after approximately 6 to 9 days, the mice were randomly assigned into two treatment groups. One group r eceived intratumoral injection of 100 μg pcDNA3. And the other graup received as recombinant plasmid. Tumor growth was determined by measuring two perpendicular diameters every three days and volume was calculated, lmmunohistochemlstry was performed to detect the expression of As protein in situ. CD31 staining was performed to assess tumor vascularity. TUNEL analysis was performed to assess apoptosis of the tumor cells. RESULTS： Tumor appeared about 4 days after C6 cells implantation. The average volume of tumors in mice treated with As gene therapy was significantly smaller than those in the control group （P〈 0.05, 1148.03±191.51mm^3 versus 1519.88±256.54mm^3） 15 days after treatment. The vascularity of the tumors treated with As was significantly suppressed when it was compared with those treated with empty vectors. More apoptotic cells were detected in tumors treated with As gene therapy. A significant increase in apoptotic index （AI） was found in this group.CONCLUSIONS： Angiostatin gene therapy inhibits tumor angiogenesis, induces the apoptosis of glioma cells and retards the growth of subcutaneous gliomas in nude mice.

关 键 词：胶质瘤 血管抑素 裸鼠 血管生成 凋亡 

分 类 号：R739.41[医药卫生—肿瘤]