【摘要】 目的:探讨分化抑制因子1(inhibitor of dif erentiation 1,Id1)在结肠癌血管生成中的作用及其可能机制。方法:将Id1过表达重组载体质粒转染结肠癌HT-29细胞后,应用实时荧光定量PCR和蛋白质印迹法检测细胞中缺氧诱导因子(hypoxia-inducible factor,HIF)-1α和血管内皮生长因子(vascular endothelial growth factor,VEGF)m RNA及蛋白的表达。应用Id1过表达的HT-29细胞培养上清液培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)24 h后,通过管状形成实验检测HUVECs管状形成的能力。裸鼠成瘤实验观察Id1过表达的HT-29细胞在裸鼠皮下的成瘤能力,免疫组织化学法检测裸鼠移植瘤组织中的微血管密度(microvessel density,MVD)。结果:Id1过表达组HT-29细胞中HIF-1α和VEGF m RNA及蛋白的表达水平均明显高于阴性对照组(转染空载体)和空白对照组(不进行任何转染),差异有统计学意义(P<0.05)。用Id1过表达的HT-29细胞培养上清液培养HUVECs,其管状结构数量明显高于阴性对照组和空白对照组(P<0.05)。Id1过表达的HT-29细胞在裸鼠皮下的成瘤能力和MVD数明显高于阴性对照组和空白对照组(P<0.05)。结论:Id1可能通过调节HIF-1α和VEGF的表达,促进结肠癌HT-29细胞的血管生成能力。更多还原

【Abstract】 Objective:To explore the role and the possible mechanism of inhibitor of differentiation 1(Id1) in angiogenesis of colon cancer.Methods:After transfection with Id1 over-expression vector plasmid,the expression levels of hypoxia-inducible factor-1α(HIF-1α) and vascular endothelial growth factor(VECF) mRNAs and proteins in HT-29 cells were detected by real-time fluorescent quantitative PCR and Western blotting,respectively.The ability of tube formation of human umbilical vein endothelial cells(HUVECs) after culture with supernatant of HT-29 cells with Id1 overexpression for 24 h was observed by tube formation experiment.The tumorigenesis of HT-29 cells with Id1 over-expression in nude mice was observed,and the microvessel density(MVD) of transplanted tumor tissues was detected by immunohistochemistry,.Results:The expression levels of HIF-1α and VECF mRNAs and proteins in HT-29 cells with Idl over-expression were higher than those in the HT-29 cells transfected with empty vector(negative control) and the HT-29 cells without any transfection(blank control)(all P < 0.05).The number of tube formation of HUVECs after culture with supernatant of HT-29 cells with Idl over-expression was higher than those of the negative control and the blank control groups(both P < 0.05).The ability of tmorigenesis and the number of MVD in Idl overexpression group were both higher than those in the negative control and the blank control groups(all P < 0.05).Conclusion:Idl may promote the angiogenesis of colon cancer by regulating the expressions of HIF-1α and VECF.更多还原