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山东大学学报(医学版) ›› 2014, Vol. 52 ›› Issue (2): 1-5.doi: 10.6040/j.issn.1671-7554.0.2013.224

• 基础医学 •    下一篇

白血病细胞HL-60诱导分化后LMP/TAP的表达变化及其意义

方敏1,毕可红2,姜国胜3,朱传升2   

  1. 1.山东大学医学院, 济南 250012; 2.山东大学附属千佛山医院血液内科, 济南 250014;
    3.山东省医学科学院基础医学研究所, 济南 250062
  • 收稿日期:2013-03-25 出版日期:2014-02-10 发布日期:2014-02-10
  • 通讯作者: 毕可红, E-mail:bikehong11@163.com
  • 基金资助:

    国家自然科学基金(30771103);山东省自然科学基金(Y2008C165)

Changes of the LMP/TAP gene expressions and its significance in leukemia cells HL-60 after induced differentiation

FANG Min1, BI Ke-hong2, JIANG Guo-sheng3, ZHU Chuan-sheng2   

  1. 1. School of Medicine, Shandong University, Jinan 250012, China;
    2. Department of Hemotology, Qianfoshan Hosipital Affiliated to Shandong University, Jinan 250014, China;
    3. Institute of Basic Medicine, Shandong Academy of Medical Sciences, Jinan 250062, China
  • Received:2013-03-25 Online:2014-02-10 Published:2014-02-10

摘要:

 目的  研究急性髓系白血病细胞HL-60诱导分化后抗原处理相关转运体蛋白(TAP)及低分子量蛋白酶体(LMP)在该细胞模型中的表达变化及其意义。方法   本实验分对照组、ATRA诱导组和PMA诱导组;Wright-Gimesa染色观察HL-60细胞形态,流式细胞术检测细胞表面标记CD11b、CD14的表达,RT-PCR检测LMP/TAP在mRNA水平的表达,Western blotting检测细胞TAP2基因的蛋白表达。结果   0.8μmol/L全反式维甲酸(ATRA)和50nmol/L乙酸肉豆蔻佛波酯(PMA)分别作用HL-60细胞72h后可显著抑制细胞增殖,并诱导细胞向成熟粒细胞及巨噬细胞方向分化; CD11b和CD14表达水平均明显升高(P均<0.05);TAP/LMP的mRNA表达水平及TAP2 的蛋白表达水平均升高(P均<0.05)。结论    白血病来源的单核样细胞具有一定的免疫功能表型,为进一步研究LMP/TAP在白血病免疫逃逸中的作用奠定基础。

关键词: 全反式维甲酸, 低分子量蛋白酶体, HL-60细胞, 乙酸肉豆蔻佛波酯, 抗原处理相关转运体蛋白, 诱导分化

Abstract:

Objective   To investigate the expression and significance of low molecular weight poly-peptide (LMP) and transporter associated with antigen processing (TAP) in leukemia cells HL-60 after induced differentiation. Methods  The cells were divided into the control group, ATRA-induced goup and PMA-induced group. Cell morphology was observed by Wright-Giemsa staining. The expressions of cell surface markers CD11b and CD14 were measured by flow cytometry. TAP1, TAP2, LMP2 and LMP7 mRNA were measured by RT-PCR, and TAP2 proteins were determined by Western blotting. Results  After treated for 72 hours, 0.8μmol/L all-trans retinoic acid (ATRA) and 50nmol/L phorbol myristate acetate (PMA) restrained the HL-60 cell proliferation and induced the cells to differentiate into the mature granulocyte and mononuclear macrophages; the expression levels of CD11b and CD14 significantly increased (all P<0.05); there were significant changes in the expressions of TAP1, TAP2, LMP2, and LMP7 mRNA (all P<0.05); the protein expression level of TAP2 was obviously decreased (P<0.05). Conclusion  Leukemia-derived mononuclear cells have certain immune function phenotype, which lay the foundation for further study of LMP/TAP in leukemia immune escape.

Key words: Induced differentiation, Transporter associated with antigen processing, Phorbol myristate acetate, Low molecular weight poly-peptide, HL-60 cell, All-trans retinoic acid

中图分类号: 

  • R733.7
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