Journal of International Oncology ›› 2017, Vol. 44 ›› Issue (4): 246-250.doi: 10.3760/cma.j.issn.1673-422X.2017.04.002

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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 Jinan-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, Jinan 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)

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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