目的:探讨低氧诱导因子(HIF)1α在紫外线诱导血管内皮细胞生长因子(VEGF)分泌中的调节作用。方法:体外培养人永生化角质形成细胞株(HaCaT),用不同剂量的UVB辐射,在照射后不同时间收集细胞,采用ELISA方法检测上清中VEGF蛋白含量。利用Lentivectors慢病毒系统建立针对HIF1α基因HaCaT(RNAi)稳转细胞株。30mJ/cm2UVB照射siRNAHIF1α稳定HaCaT细胞组和正常HaCaT细胞组,24h上清检测VEGF的蛋白表达。结果:UVB可增强VEGF表达.并呈剂量依赖性和时间依赖性。与正常HaCaT细胞组比,siRNAHIF1α稳定HaCaT细胞组VEGF的蛋白表达量明显降低。结论:siR—NAHIF1α可以抑制UVB诱导HIF1α目的基因的表达。
Objective: To determine the role of HIF1 in UV-induced expression of its target genes vascular endothelial growth factor (VEGF). Methods: HaCaT cells were irradiated with UVB. ELISA was performed to detect VEGF protein level at different time points after treatments. HaCaT and HaCaT(HIF1 RNAi) cells (70% confluence) were starved by replacing the medium with 0.1% FBS DMEM and culturing for 24 h. The cells were irradiated with UVB(30 mJ/cm^2) and subsequently cultured for 24 h. The VEGF expressions were detected. HaCaT (HIF1 RNAi) cells were strongly attenuate VEGF levels after UVB irradiation(P 〈 0.05) when compared with HaCaT cells. Results: Compared with control groups, UVB was able to induce VEGF protein expression in a dose and time dependent manner in HaCaT cells (P 〈 0.05). HIF1 RNAi can attenuate VEGF levels after UVB irradiation(P 〈 0.05). Conclusion: UVB can enhance the expression of VEGF. HIF1 can attenuate VEGF levels after UVB irradiation.