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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。
Objective: To investigate the role of inhibitor of dif erentiation 1 (Id1) in angiogenesis of colon carcinoma and its possible mechanism. Methods: After transfection of Id1 overexpression vector into HT-29 cells, the expression of hypoxia-inducible factor (HIF-1α) and vascular endothelial growth factor (VEGF) were detected by real-time fluorescence quantitative PCR and Western blotting. (vascular endothelial growth factor, VEGF) m RNA and protein expression. HUVECs were cultured for 24 h in culture supernatant of HT-29 cells with Id1 overexpression. The tubular formation ability of HUVECs was detected by tubular formation assay. The tumorigenicity of Id-1-overexpressing HT-29 cells in nude mice was observed by tumorigenicity assay in nude mice. Microvessel density (MVD) was detected by immunohistochemistry in nude mice. Results: The expression levels of HIF-1α, VEGF m RNA and protein in HT-29 cells with Id1 overexpression group were significantly higher than those in the negative control group (transfected with empty vector) and blank control group (without any transfection), the difference was Statistical significance (P <0.05). HUVECs were cultured in HT-29 cell culture supernatants with Id1 overexpression. The number of tubular structures was significantly higher than that of negative control group and blank control group (P <0.05). The tumorigenicity and MVD of Id-1-overexpressing HT-29 cells in nude mice were significantly higher than those in negative control group and blank control group (P <0.05). Conclusion: Id1 may promote the angiogenesis of colon cancer HT-29 cells by regulating the expression of HIF-1α and VEGF.