摘要目的 利用显微图像分析法研究肿瘤新生血管相关模型.方法 以人脐静脉内皮细胞和胃癌细胞系建立体外小管形成模型与血管拟态形成模型,以鸡的受精卵建立鸡胚尿囊膜活体血管形成模型,观察抑癌基因IRX1对血管形成和血管拟态形成的影响.显微镜所摄数字图片通过Image Pro Plus(IPP)专业图像软件进行分析.通过计数点和线长度分析体外小管形成;通过吸光度法测量血管拟态的PAS阳性物质;通过计数法和角度测量法计算鸡胚尿囊膜血管数目.结果 IRX1转基因组与空载体组及空白对照组相比,均显示了抑制血管形成能力,其小管形成数量三组分别为(12.80±3.83)、(29.00±5.34)和(28.20±4.32)个(P<0.01),三组的小管连接点分别为(13.20±2.59)、(25.00±2.24)和(24.60±3.21)个(P<0.01),三组的小管长度分别为(821.5±12.5)、(930.9±13.5)和(948.4±18.1)μm(P=0.022).IRX1基因转染组的PAS阳性物质吸光度值(3606±363)显著低于空载体组(14 200±1251)与空白对照组(15 043±1220,P<0.01).IRX1基因转染组的鸡胚尿囊膜血管形成数量明显少于空载体组与空白对照组,三组角测量血管数量分别为(6.41±2.60)、(10.27±2.65)和(9.18±1.99)个(P<0.01).结论 体外血管形成、血管拟态模型和鸡胚活体血管形成模型研究均显示,IRX1基因转染可以明显抑制新生血管形成;上述模型适用于以血管为靶点的基因干预或者药物筛选研究;专业图像分析软件在肿瘤血管形成的定量分析研究中可发挥一定的作用.

abstractsObjective To establish experimental models for tumor neovascularization and to apply quantitative digital imaging analysis in the study. Methods An endothelial tube formation model was established by human umbilical vein endothelial cells (HUVECs). A vasculogenic mimicry model was established by SGC-7901 gastric cancer cell line. Fertilized eggs were used to establish a chorioallantoic membrane angiogenesis model. Using gene transfection experiment, IRX1 tumor suppressor gene was chosen as a therapeutic target. Image Pro Plus (IPP) analysis software was used for digital vascular images analysis with parameters including points, lines, angles and integral absorbance (IA) for the tubular formation or vasculogenic mimicry.Results Digital image analysis by IPP showed that HUVEC tubular formation was significantly inhibited in IRX1 transfectant, compared with controls. The tubular numbers in three groups were 12.80±3.83, 29.00±5.34 and 28.20±4.32(P<0.01).The connection points of tubules in three groups were 13.20±2.59, 25.00±2.24 and 24.60±3.21(P<0.01). The tubular lengths of three groups were (821.5±12.5), (930.9±13.5)and(948.4±18.1)μm(P=0.022). The IA values of PAS stain in three groups were 3606±363, 14 200±1251 and 15 043±1220 (P<0.01). In chick chorioallantoic membrane model, the angular numbers of tubules in three groups were 6.41±2.60, 10.27±2.65 and 9.18±1.99(P<0.01). Conclusions The endothelial tube formation model, vasculogenic mimicry model and chorioallantoic membrane angiogenesis model are useful for gene therapy and drug screening with targeting neoplastic vascularization. Professional image analysis software may greatly facilitate the quantitative analysis of tumor neovascularization.