Arresten对人脐静脉内皮细胞增殖和凋亡影响的研究

Inhibition of arresten on proliferation and apoptosis of Huvec cells

目的:观察血管生成抑制因子Arresten对人脐静脉内皮细胞Huvec增殖和凋亡的影响,探讨VEGFR在其中的作用。方法:应用MTT法检测不同浓度的Arresten对Huvec细胞增殖的影响;流式细胞仪检测Arresten对Huvec细胞凋亡的影响;半定量RT-PCR和蛋白质印迹法分别检测VEGFR mRNA水平及VEGFR蛋白水平的表达。结果:MTT比色法显示,Arresten对Huvec细胞增殖具有抑制作用,随着浓度的增加,Arresten对Huvec细胞增殖抑制作用也增强;但当浓度增加到800 ng/L后其增殖抑制率不再提高,而此浓度时Huvec细胞的凋亡率达到峰值的(59±0.3)%。经过Ar-resten处理,Huvec细胞的VEGFR mRNA表达及VEGFR蛋白表达均显著下降。结论:Arrest-en通过降低Huvec细胞的VEGFR表达而抑制Huvec细胞增殖并促进Huvec细胞凋亡,这可能是Arresten抑制血管生成的机制之一。

OBJECTIVE： To investigate the inhibition of arresten on proliferation and apoptosis of Huvec cells, and its relation with VEGFR. METHODS： MTT was used to measure the proliferation of Huvec cells, the flow cytometry assay （FCM） was used to detect the apoptosis of Huvec cells; VEGFR mRNA and VEGFR protein was measured by semi- quantitative RT PCR and Western blot, respectively. RESULTS： The proliferation of Huvec ceils was inhibited by arresten, and with the increase of concentration of arresten, the cell viability of Huvec cell lines decreased gradually. However, when the concentration of arresten was above 800 ng/mL, Huvec cells were resistant to arresten. After incubation with arresten （800 ng/mL） for 24 h, the rate of apoptosis in Huvec cells reached to （59±0.3）%. The expressions of VEGFR mRNA and VEGFR protein in Huvec cells decreased after being incubated with arresten. CONCLUSION： Arresten inhibits the proliferation of Huvec cells and induces the apoptosis, and in this course VEGFR plays an important role.