国际肿瘤学杂志

国际肿瘤学杂志 ›› 2017, Vol. 44 ›› Issue (4): 246-250.doi: 10.3760/cma.j.issn.1673-422X.2017.04.002

• 论著 • 上一篇    下一篇

白血病细胞诱导分化过程中nm23-H1表达变化及其分子调控机制

田静,袁小芬,韩杨,任霞,张之勇,郭强,李莲莲,张晓瑜,张振,李霞,姜国胜   

  1. 250062 济南大学 山东省医学科学院医学与生命科学学院（田静、韩杨）；山东省医学科学院基础医学研究所 山东省现代医用药物与技术重点实验室 山东省医药卫生肿瘤免疫与中药免疫重点实验室（田静、韩杨、任霞、张之勇、郭强、李莲莲、张晓瑜、张振、李霞、姜国胜）；山东中医药大学第二附属医院预防保健中心（袁小芬)

  • 出版日期:2017-04-08 发布日期:2017-05-09
  • 通讯作者: 姜国胜 E-mail:jiangguosh@163.com
  • 基金资助:

    国家自然科学基金(81573467)；山东省自然科学基金（ZR2015HM014)；山东省自主创新及成果转化专项(2014CGZH1313,2015ZDJS04003)；山东省泰山学者工程（ts201511075）；山东省科技发展计划（2013YD18031）

Expression change of nm23-H1 in differentiation process of leukemia cells and its molecular mechanisms

Tian Jing,Yuan Xiaofen,Han Yang,Ren Xia,Zhang Zhiyong,Guo Qiang,Li Lianlian,Zhang Xiaoyu,Zhang Zhen,Li Xia,Jiang Guosheng   

  1. School of Medicine and Life Sciences, University of Ji′nan-Shandong Academy of Medical Sciences; Institute of Basic Medicine, Shandong Academy of Medical Sciences, Key Laboratory for Modern Medicine and Technology of Shandong Province, Key Medical Laboratory for Tumor Immunology of Shandong Province, Ji′nan 250062, China
  • Online:2017-04-08 Published:2017-05-09
  • Contact: Jiang Guosheng E-mail:jiangguosh@163.com
  • Supported by:

    National Natural Science Foundation of China (81573467); Natural Science Foundation of Shandong Province of China (ZR2015HM014); Shandong Key Project for Transformation of Results with Independent Innovation(2014CGZH1313, 2015ZDJS04003); Project for Laureate of Taishan Scholar of Shandong Province of China (ts201511075); Key Science and Technology Program of Shandong Province of China (2013YD18031)

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摘要： 目的 探讨在白血病细胞HL-60分化过程中nm23-H1、c-Myc、粒细胞集落刺激因子（G-CSF）的作用机制及相互关系，为临床治疗提供理论依据和治疗靶点。方法 CCK-8实验测定全反式维甲酸（ATRA）作用下的靶细胞增殖情况，瑞士-吉姆萨染色观察细胞形态学变化。反转录-聚合酶链反应（RT-PCR）或Western blotting测定nm23-H1、c-Myc和G-CSF的基因、蛋白表达水平。酶联免疫吸附试验（ELISA）测定ATRA诱导HL-60细胞分化过程中培养上清G-CSF水平的变化。结果 分别使用药物浓度为0.25、0.50、1.00、2.00和4.00 μmol/L的ATRA诱导HL-60细胞72 h，细胞增殖抑制率分别为（43.00±0.08）%、（49.55±2.30）%、（58.90±6.70）%、（64.60±4.70）%、（72.70±3.20）%，差异有统计学意义（F=69.887，P=0.000）。在ATRA诱导下，细胞形态学证实HL-60细胞向成熟粒细胞方向分化。nm23-H1的mRNA相对表达量由0.79±0.03逐渐降低到0.52±0.09，差异有统计学意义（F=9.679，P=0.002），蛋白相对表达量由1.54±0.12逐渐降低到0.40±0.04，差异有统计学意义（F=90.140，P=0.000）。c-Myc的mRNA相对表达量由0.95±0.05逐渐降低到0.46±0.05，差异有统计学意义（F=27.900，P=0.000），蛋白相对表达量由1.12±0.11逐渐降低到0.14±0.03，差异有统计学意义（F=115.500，P=0.000）。而G-CSF的mRNA相对表达量由0.26±0.07逐渐升高到0.55±0.09，差异有统计学意义（F=7.817，P=0.004），细胞分泌水平由（13.67±7.51）pg/ml逐渐升高到(128.30±25.77)pg/ml，差异有统计学意义（F=28.020，P=0.000）。结论 nm23-H1可能通过与c-Myc和G-CSF相互作用而发挥其诱导白血病HL-60细胞分化的作用

关键词: font-family: 宋体, mso-bidi-font-family: 宋体, mso-font-kerning: 1.0pt, mso-ansi-language: EN-US, mso-fareast-language: ZH-CN, mso-bidi-language: AR-SA">白血病, 细胞分化, 粒细胞集落刺激因子

Abstract: Objective To detect the mechanisms of nm23-H1, c-Myc, granulocyte colony-stimulating factor (G-CSF) in differentiation of leukemia cells HL-60 and their correlation, and provide theory basis and treatment targets for clinical treatment. Methods We chose CCK-8 assay to detect the proliferation change of the HL-60 cells induced by optimal concentration of all-trans-retinoic acid   (ATRA). Wright Giemsa staining was used to observe cell morphological changes. The gene or protein expressions of nm23-H1, c-Myc and G-CSF were detected by reverse transcription-polymerase chain reaction (RT-PCR) or Western blotting assay, and the G-CSF secretion level in supernatants of ATRA-inducing HL-60 cells was measured by enzyme linked immunosorbent assay (ELISA). ResultsThe cell proliferation inhibition rates were respectively (43.00±0.08)%, (49.55±2.30)%, (58.90±6.70)%, (64.60±4.70)%, (72.70±3.20)%, after HL-60 cells were induced by ATRA with drug concentrations of 0.25, 0.50, 1.00, 2.00 and 4.00 μmol/L for 72 hours, with a significant difference (F=69.887, P=0.000). After induced by ATRA, cell morphology confirmed that HL-60 cells were differentiated into mature granulocytes. The mRNA expression of nm23-H1 decreased gradually from 0.79±0.03 to 0.52±0.09. The difference was statistically significant (F=9.679, P=0.002). The relative expression of protein decreased from 1.54±0.12 to 0.40±0.04. The difference was statistically significant (F=90.140, P=0.000). The relative expression of c-Myc mRNA decreased from 0.95±0.05 to 0.46±0.05. The difference was statistically significant (F=27.900, P=0.000). The relative expression of protein decreased from 1.12±0.11 to 0.14±0.03. The difference was statistically significant (F=115.500, P=0.000). Moreover, the mRNA expression of G-CSF increased gradually from 0.26±0.07 to 0.55±0.09. The difference was statistically significant (F=7.817, P=0.004). The level of cell secretion increased from (13.67±7.51)pg/ml to (128.30±25.77)pg/ml. The difference was statistically significant (F=28.020, P=0.000). Conclusionnm23-H1 probably plays an important role in inducing differentiation of HL-60 cells by interacting with the expressions of c-Myc and G-CSF

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