全反式维甲酸对HL-60细胞VEGF表达与分泌影响的分子机制研究  

Mechanism of VEGF expression and secretion in HL-60 cells induced by all-tram refinoic acid

作　　者：范华[1] 温培娥[1] 杨炜华[1] 任霞[1] 赵海涛[1] 乔高娟[1] 唐天华[1] 任海全[1] 姜国胜[1] 

机构地区：[1]山东省医学科学院基础医学研究所山东省医药卫生肿瘤免疫与中药免疫重点实验室,济南250002

出　　处：《国际肿瘤学杂志》2009年第4期316-319,共4页Journal of International Oncology

基　　金：国家自然科学基金资助项目（30771103）;山东省自然科学基金资助项目（Y2008C165）;山东省卫生系统杰出学科带头人和中青年重点科技人才基金资助项目（鲁卫科教国合发[2008]2号）

摘　　要：目的探讨人髓系白血病细胞株HL-60在诱导分化过程中血管内皮生长因子（VEGF）表达及分泌水平变化的分子机制。方法四甲基偶氮唑蓝（MTr）法检测HL-60细胞被全反式维甲酸（ATRA）诱导分化后的增殖水平变化，流式细胞术测定HL-60细胞CD11b表达和细胞周期的改变，半定量逆转录聚合酶链式反应（RT—PCR）检测HL-60细胞VEGF、c-myc、缺氧诱导因子（HIF）-1α、基质金属蛋白酶（MMP）-9和MMP-2 mRNA表达水平，应用酶联免疫吸附试验（ELISA）法检测药物作用前后HL-60细胞培养上清液中VEGF蛋白的含量。结果经ATRA诱导分化后，HL-60细胞增殖水平明显降低，CD11b表达水平明显升高，出现粒系定向分化趋势，且分化程度明显升高（P〈0．05），c-myc与VEGF mRNA表达水平均明显下调（P〈0．05），HIF-1α mRNA表达水平则明显升高（P〈0．05）。用MMP-2及MMP-9抑制因子I阻断HL-60细胞MMP-9和MMP-2表达后，细胞培养上清中VEGF蛋白分泌量明显降低（P〈0．05）。结论HL-60细胞被诱导分化过程中VEGF的表达与c-myc表达正相关，与HIF-1α表达负相关。MMP-9、MMP-2可能是调控HL-60细胞分泌VEGF的主要原因之一。Objective To explore molecular mechanism of expression of vascular endothelial growth factor （VEGF） mRNA and secretion of VEGF protein in HL-60 cells induced by all-trans refinoic acid （ATRA）. Methods MTT method was used to detect the proliferation of HL-60 cells induced by ATRA,ceU cycle and CD11 b expression in HL-60 cells were detected by flow cytometry. Expression of VEGF, c-myc, hypoxia-inducible factor （HIF）-1α, matrix metalloproteinase （MMP）-9 and MMP-2 mRNA were detected by semi-quantitative RT-PCR. VEGF protein in HL-60 ceils culture supematant was measured by ELISA before and after being induced by ATRA. Results After treatment with ATRA,the proliferation of HL-60 cells was obviously inhibited, CD11b expression increased, trend of granulocyte directional differentiation emerged, and differentiation degree was increasd （ P 〈 0. 05 ） ;expression level of c-my and VEGF mRNA was down-regulated （ P 〈 0. 05 ）, but expression level of HIF-1α mRNA was up-regulated （ P 〈 0. 05 ）. VEGF protein level in HL-60 cells culture supernatant was decreased by blocking the expression of MMP-9 or MMP-2（ P 〈0. 05）. Conclusion VEGF expression has positive correlation with c-myc expression,but has negative correlation with HIF-1α expression. MMP-9 and MMP-2 may be the main factors regulating VEGF secretion in HL-60 cells.

关 键 词：维甲酸 白血病 血管内皮生长因子类 基因 myc 基质金属蛋白酶类 

分 类 号：R73-3[医药卫生—肿瘤]