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，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 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:2012-11-08 Published: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