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白血病细胞HL-60诱导分化后LMP/TAP的表达变化及其意义

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

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【作者】 方敏; 毕可红; 姜国胜; 朱传升;

【Author】 FANG Min;BI Ke-hong;JIANG Guo-sheng;ZHU Chuan-sheng;School of Medicine,Shandong University;Department of Hemotology,Qianfoshan Hosipital Affiliated to Shandong University;Institute of Basic Medicine Shandong Academy of Medical Sciences;

【机构】 山东大学医学院; 山东大学附属千佛山医院血液内科; 山东省医学科学院基础医学研究所;

【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50 nmol/L乙酸肉豆蔻佛波酯(PMA)分别作用HL-60细胞72 h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。

【Abstract】 Objective To investigate the expression and significance of lowmolecular 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 flowcytometry. 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. 8 μmol / L all-trans retinoic acid( ATRA) and 50 nmol / 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.

【基金】 国家自然科学基金(30771103);山东省自然科学基金(Y2008C165)
  • 【文献出处】 山东大学学报(医学版) ,Journal of Shandong University(Health Sciences) , 编辑部邮箱 ,2014年02期
  • 【分类号】R733.7
  • 【被引频次】1
  • 【下载频次】80
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