国际肿瘤学杂志

国际肿瘤学杂志››2012,Vol. 39››Issue (11): 873-877.

• 论著 •上一篇下一篇

HL-60细胞诱导分化后血管内皮生长因子表达变化分子机制的探讨

李燕梅, 宋冠华, 温培娥, 任霞, 毕可红, 姜国胜

  1. 250062济南，betway.com 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山东省罕见病重点实验室(李燕梅、宋冠华、温培娥、任霞、唐天华、姜国胜)；山东省千佛山医院血液科(毕可红)
  • 出版日期:2012-11-08发布日期:2012-11-16
  • 通讯作者:姜国胜，E-mail：jianggsh@hotmail.com E-mail:jianggsh@hotmail.com
  • 基金资助:

    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（ZR2011HL050）；山东省科技攻关重点项目（2007GG20022023）

Study on molecular regulation mechanism of VEGF in HL-60 cells after induced differentiation

LI Yan-Mei, SONG Guan-Hua, WEN Pei-E, REN Xia, BI Ke-Hong, JIANG Guo-Sheng

  1. Institute of Basic Medicine,Shandong Academy of Medical Science,Key Medical Laboratory for Tumor Immunology and Chinese Medicine Immunology of Shandong Province,Key Laboratory of Rare&Uncommon Diseases of Shandong Province, Jinan 250062,China
  • Online:2012-11-08Published:2012-11-16
  • Contact:Corresponding author: JIANG Guo-sheng, E-mail: jianggsh@hotmail.com E-mail:jianggsh@hotmail.com

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摘要：目的 探讨全反式维甲酸（ATRA）诱导HL-60细胞分化模型中血管内皮生长因子（VEGF）表达调控的分子机制，为白血病的抗血管新生治疗提供新的靶点。方法 Wright-Gimesa染色观察细胞形态，硝基四唑氮蓝(NBT)还原实验检测HL-60细胞分化，逆转录-聚合酶链反应（RT-PCR）、蛋白质印记（Western blot）分别从mRNA和蛋白水平检测细胞VEGF、Stat3、c-myc的表达变化。结果 1 μmol ATRA作用于HL-60细胞96 h后可明显抑制细胞增殖，并出现明显的分化现象，倒置显微镜下可观察到细胞向成熟粒细胞方向分化；NBT阳性率为82.59% （t=﹣24.157，P＜0.01）；VEGF mRNA表达水平下降（t=7.339，P＜0.05）、STAT3 mRNA表达水平下降（t=3.667，P＜0.05）、c-myc mRNA表达水平下降（t=6.858，P＜0.05）。VEGF蛋白表达水平下降（t=3.386，P＜0.05）、STAT3蛋白表达水平下降（t=4.074，P＜0.05）、c-myc蛋白表达水平下降（t=3.333，P＜0.05）。结论 在ATRA诱导HL-60细胞分化的模型中VEGF的表达水平随诱导分化的进程而下降，其下调机制可能与STAT3、c-myc具有相关性。

关键词:维甲酸,血管内皮生长因子类,HL-60细胞

Abstract:Objective To study the molecular regulation mechanism of VEGF in the model of ATRA induced differentiation in HL-60 cells, and to provide new targets for leukemia anti-angiogenic therapy. Methods The morphology was observed by Wright-Gimesa staining; HL-60 cells differentiation was detected by NBT reduction experiment. VEGF,STAT3,c-myc mRNAs were measured by reverse transcription-PCR; VEGF,STAT3 and c-myc proteins were determined by Western blot. Results The proliferation of HL-60 cells was inhibited obviously by ATRA(1 μmol/L) with the induction of differentiation, NBT positive rate was 82.59% (t=﹣24.157，P <0.01); VEGF mRNA(t=7.339，P＜0.05), STAT3 mRNA (t=3.667，P＜0.05) and c-myc mRNA（t=6.858，P＜0.05） were all down-regulated. VEGF protein（t=3.386，P＜0.05）, STAT3 protein（t=4.074，P＜0.05）and c-myc protein （t=3.333，P＜0.05）were all down-regulated. Conclusion VEGF expression level is reduced with the procession of differentiation of HL-60 cells, which may be largely correlated with the down regulation of STAT3 and c-myc.

Key words:Tretinoin,Vascular endothelial growth factors,HL-60 cells