VEGF在HaCaT细胞中的自分泌作用

Autocrine effect of vascular endothelial growth factor on the proliferation of HaCaT cells

目的:检测VEGF在皮肤角质形成细胞系HaCaT细胞中可能的自分泌作用。方法:用MTT法,检测不同浓度外源性VEGF165(0,1,5,10,25,50,100 ng/ml)和不同浓度VEGF特异性抑制剂阿瓦斯丁(0,0.063,0.125,0.25,0.50,1.0,2.0 mg/ml)作用下HaCaT细胞增殖的变化;用细胞迁移实验,检测不同浓度VEGF165和0.5 mg/ml阿瓦斯丁作用下HaCaT细胞迁移的变化;蛋白免疫印迹检测10 ng/ml VEGF165和0.5mg/ml阿瓦斯丁作用下HaCaT细胞ERK1/2的磷酸化情况。结果:VEGF165剂量依赖性地促进HaCaT细胞的增殖和迁移能力;而阿瓦斯丁剂量依赖性地抑制其增殖能力,0.50 mg/ml阿瓦斯丁可以显著性降低其迁移能力;10 ng/ml VEGF165显著性促进HaCaT细胞ERK1/2的磷酸化,而对0.5 mg/ml阿瓦斯丁预处理过的HaCaT细胞的ERK1/2磷酸化没有促进作用。结论:VEGF在皮肤角质形成细胞系HaCaT中存在着自分泌作用。

Objective： To determine the autocrine effect of vascular endothelial growth factor （VEGF） on epidermal keratinocytes HaCaT cells. Methods： Cultured HaCaT cells were treated with various concentrations of VEGF16s （0, 1,5,10, 25,50,100 ng/ml） or Avastin （0, 0. 063, 0. 125,0.25,0.50,1.0,2.0 mg/ml） in vitro. HaCaT cell proliferation was determined by MTT assay and the cell migration was measured by migration assay. The effect of VEGF16s（10 ng/ml） on phosphorylation of ERK1/2 was detected in HaCaT cells pretreated or not pretreated with Avastin （0. 5 mg/ml）. Results： VEGF enhanced the proliferation and migration of HaCaT cells in a dose-dependent manner, while Avastin inhibited the effects of VEGF also in a dose-dependent manner. VEGF16s（10 ng/ml） induced the phosphorylation of ERK1/2 in HaCaT cells,but which was blocked by Avastin （0. 5 mg/ml）. Conclusion： An autocrine function of VEGF exits in HaCaT cells.