【摘要】 目的观察血管生成素-1(Angiopoietin-1,Ang-1)蛋白对无血清DMEM培养基培养的结肠癌细胞(HCT-8)存活率的影响,探讨其与PI3-’kinase/Akt通路的关系。方法将不同浓度(0、0.05、0.2、0.4、0.82、.0 mg/L)的Ang-1蛋白作用于HCT-8细胞,用MTT法检测细胞增殖,根据实验结果选定Ang-1蛋白的后续实验浓度,加入Ang-1及LY294002,应用Western blotting蛋白免疫印记法分析相关蛋白(Tie-2、PI3K、Akt)的变化。结果 Ang-1(0.05、0.2、0.4、0.82、.0 mg/L)+DMEM组与无血清DMEM培养基组比较,Tie-2、PI3K、Akt三种蛋白在HCT-8细胞中的表达均增强(P<0.01,P<0.01,P<0.01),DMEM+Ang-1+LY294002组三种蛋白的表达均减弱(P>0.05,P<0.01,P<0.01)。结论较低浓度(0.05 mg/L)的血管生成素-1蛋白在结肠癌细胞中即有抗凋亡作用,并且随着剂量的增加抗凋亡作用平稳,其诱导凋亡的机理可能与Tie-2/PI3-’kinase/Akt调节的通路有关,应用该途径的抑制剂LY294002可抑制结肠癌细胞的生长,实现抗肿瘤作用。更多还原

【Abstract】 Objective To observe the effect of Angiopoietin-1 on livability of HCT-8 with no-serum low-sugar culture medium,to study the relation between Ang-1 and the access of PI3K/Akt.Methods Different dose of Ang-1 were added to HCT-8 cell lines,then MTT was applied to evaluate the degree of cell produce.Then HCT-8 were cultured in the medium containing suitable Ang-1 and LY294002,Tie-2,PI3K and Akt were detected by western blotting.Results Tie-2、PI3K、Akt are increasing from Ang-1+DMEM group to no-serum low-sugar culture medium group(P<0.01,P<0.01,P<0.01),Ang-1+DMEM+LY294002 are reducing(P>0.05,P<0.01,P<0.01).Conclusion Levels of Ang-1 for metastatic colon cancer cells are resisting the apoptosis and have a dose of his physical dependence.The mechanism of apoptosis may have relation with the throttlechannel of Tie-2/PI3’-kinase /Aktmay,the application of the ways by inhibitors LY294002 for metastatic colon cancer cells may restrain growth and resistance the tumor.更多还原