摘要
本研究旨在应用人脐静脉内皮细胞(HUVEC)及K562细胞探讨干扰素-α2b在慢性髓系白血病(CML)中抗血管生成作用。用ELISA法检测K562细胞系培养上清液中VEGF和bFGF水平;应用real-time RT-PCR法检测103、102、10U/ml IFN-α2b作用K562细胞24、36、48小时VEGF、bFGF mRNA的表达;通过MTT法、Transwell室及体外微管形成实验研究K562细胞培养上清液及IFN-α2b对HUVEC增殖、迁移及体外分化的影响。结果表明:K562细胞系表达和分泌VEGF和bFGF,细胞培养上清液能明显促进HUVEC的增殖、迁移、体外微管形成,其作用随着细胞培养上清液浓度的增加而增强;IFN-α10U/ml作用K562细胞24、36、48小时,VEGF相对表达量为1.64±0.18、1.49±0.14、1.31±0.05,bFGF相对表达量分别为1.53±0.10、1.29±0.15、0.79±0.13(p=0.002),随着药物浓度增高,细胞VEGF、bFGF相对表达量无明显差异。结论:CML存在血管新生,K562细胞分泌和表达促血管新生因子VEGF和bFGF,促进HUVEC的增殖、迁移、体外微管形成,干扰素通过抑制HUVEC的增殖、迁移、微管形成,下调K562细胞VEGF和bFGF mRNA表达,具有一定程度的抗血管新生作用。
This study was aimed to investigate the anti-angiogenesis of IFN-α2b in chronic myeloid leukemia （CML） in vitro by using K562 cell line and human umbilical vein endothelial cells（HUVEC）. The levels of vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （bFGF） in culture supematant of K562 cells were determined by ELISA; the expressions of VEGF and bFGF mRNA after treating K562 cells with 103, 102 and 10 U/ml IFN-α2b for 24,36,48 hours were detected by real-time RT-PCR; the effects of K562 cell culture supernatant and IFN-α2b on proliferation, migration and differentiation of HUVEC in vitro were assayed by MTT, Transwell chamber and tubule formation assay respectively. The results showed that the K562 cells expressed and secreted VEGF and bFGF. The culture supernatant of K562 cells significantly promoted the proliferation, migration and tubule formation of HUVEC in vitro in a concentration-dependent manner. After treating K562 cells with IFN--α2b 10 U/mi for 24,36 and 48 hours, the expression levels of VEGF and bFGF mRNA were 1.64±0.18, 1.49 ±0.14, 1.31 ±0.05 and 1.53±0.10, 1.29 ±0.15, 0. 79 ± 0.13 respectively （p = 0. 002）, but the expression levels of VEGF and bFGF mRNA were not significantly different along with increasing of IFN-c^2b concentration. It is concluded that the angiogenesis exists in CML. The K562 cell expresses and secrets VEGF and bFGF, which promotes the proliferation, migration and differentiation of HUVEC. The IFN-α2b displays anti-angiogenesis by inhibiting the proliferation, migration and tubule formation in vitro of HUVEC and down regulating the expression of VEGF and bFGF mRNA.
出处
《中国实验血液学杂志》
CAS
CSCD
2010年第2期291-295,共5页
Journal of Experimental Hematology
基金
湖北省科技攻关项目
编号2005AA301C52
