Wang Jia-Qi, Gao You-Shui, Mei Jiong, Xue Hua-Ming, Wang Shu-Qing, Cai Xuan-Song
Department of Orthopedics, Tongji Hospital, Tongji University, Shanghai 200065, P. R. China.
Ai Zheng. 2009 Jun;28(6):581-6.
Vascular endothelial growth factor (VEGF) is a critical stimulator for angiogenesis of osteosarcoma. This study was to investigate the morphological changes in osteosarcoma xenografts in nude mice after inhibiting angiogenesis by Ad-VEGF-siRNA.
The animal model of osteosarcoma xenografts was constructed by subcutaneous inoculation of osteosarcoma MG63 cells into nude mice. The mice were randomly divided into three groups, 15 mice in each group: group A and control groups B and C. The tumor nodules were injected with Ad-VEGF-siRNA in group A, an equal dose of Ad-EGFP and PBS, respectively, in groups B and C. The volume and weight of tumors were measured every two days after injection and the growth curves were drawn. The tumor samples were observed and analyzed using HE staining. Angiogenesis was assessed by immunohistochemical staining using anti-VEGF and CD31 antibodies. The ultrastructure of vasculogenic mimicry (VM) was observed using transmission electron microscopy (TEM).
The mean volume, weight, microvessel density (MVD) and VEGF expression of tumors in Ad-VEGF-siRNA-treated group were significantly lower than those in the control groups. MVD was positively correlated to VEGF expression, with a correlation coefficient of 0.9989. The number of VM was 1.40+/-0.55 in the Ad-VEGF-siRNA group, which was significantly less than those in the two control groups.
Ad-VEGF-siRNA could effectively block angiogenesis in osteosarcoma xenografts in nude mice.
血管内皮生长因子(VEGF)是骨肉瘤血管生成的关键刺激因子。本研究旨在探讨腺病毒介导的VEGF小干扰RNA(Ad-VEGF-siRNA)抑制血管生成后裸鼠骨肉瘤移植瘤的形态学变化。
将骨肉瘤MG63细胞皮下接种于裸鼠,构建骨肉瘤移植瘤动物模型。将小鼠随机分为3组,每组15只:A组和对照组B、C组。A组瘤结节注射Ad-VEGF-siRNA,B、C组分别注射等量的Ad-EGFP和磷酸盐缓冲液(PBS)。注射后每2天测量肿瘤体积和重量并绘制生长曲线。采用苏木精-伊红(HE)染色观察和分析肿瘤样本。用抗VEGF和CD31抗体免疫组化染色评估血管生成情况。用透射电子显微镜(TEM)观察血管生成拟态(VM)的超微结构。
Ad-VEGF-siRNA治疗组肿瘤的平均体积、重量、微血管密度(MVD)和VEGF表达均显著低于对照组。MVD与VEGF表达呈正相关,相关系数为0.9989。Ad-VEGF-siRNA组VM数量为1.40±0.55,明显少于两个对照组。
Ad-VEGF-siRNA可有效阻断裸鼠骨肉瘤移植瘤的血管生成。