上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

G-CSF通过miR-146a/Smad4信号通路对HL-60白血病细胞增殖的调控作用

李昕1,2，盛贤福1,2，蔡佳翌2，徐岚1,2，沈莉菁2，钟济华2，陈芳源1,2，钟华1,2   

  1. 上海交通大学 医学院 1. 附属仁济医院南院血液科, 上海 201112； 2.附属仁济医院血液科， 上海 200127
  • 出版日期:2015-12-28 发布日期:2016-01-21
  • 通讯作者: 钟华, 电子信箱: zhh_lj@163.com。
  • 作者简介:李昕（1990—）， 女， 硕士生; 电子信箱: tiffany512@sina.com。
  • 基金资助:

    国家自然科学基金（81270626）

Regulation of the proliferation of leukemia cells HL-60 by G-CSF via the signaling pathway of miR-146a/Smad4

LI Xin1,2, SHENG Xian-fu1,2, CAI Jia-yi2, XU Lan1,2, SHEN Li-jing2, ZHONG Ji-hua2, CHEN Fang-yuan1,2, ZHONG Hua1,2   

  1. 1.Department of Hematology, Renji Hospital South Campus, Shanghai Jiao Tong University School of Medicine, Shanghai 201112, China； 2.Department of Hematology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
  • Online:2015-12-28 Published:2016-01-21
  • Supported by:

    National Natural Science Foundation of China, 81270626

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摘要：

目的  探讨粒细胞集落刺激因子（G-CSF）对白血病细胞株HL-60中miR-146a/Smad4表达水平和白血病细胞增殖活性的影响。方法  采用G-CSF (50 ng/mL) 提前48 h处理人白血病HL-60细胞株，另设G-CSF未处理组作为对照。采用CCK8法检测细胞增殖情况，流式细胞术检测细胞周期变化，实时荧光定量PCR检测 miR-146a及 Smad4 mRNA表达情况，Western blotting检测总Smad4蛋白表达。结果  G-CSF升高白血病细胞miR-146a表达，同时降低细胞 Smad4 mRNA和总Smad4蛋白的表达，提高白血病细胞的增殖活性。转染了 miR-146a过表达质粒后的白血病细胞 Smad4 mRNA和总Smad4蛋白表达均降低，而用G-CSF处理转染后的细胞，其miR-146a表达升高及Smad4表达降低得更为明显。结论  G-CSF通过调控miR-146a/Smad4信号轴，影响Smad4表达水平，刺激处于静止状态的白血病细胞进入细胞增殖阶段。

关键词: 粒细胞集落刺激因子, miR-146a/Smad4表达, 细胞增殖活性

Abstract:

Objective  To investigate the effects of granulocyte colony stimulating factor (G-CSF) on the expression level of miR-146a/Smad4 of leukemia cell line HL-60 and proliferative activity of leukemia cells. Methods  Human leukemia cell line HL-60 was treated 48 h in advance and controls were untreated leukemia cells. CCK8 was applied to detect the cell proliferation and flow cytometry was used to detect the changes of cell cycle. The mRNA expressions of miR-146a and Smad4 were detected by realtime quantitative PCR. The total protein expression of Smad4 was detected by Western blotting. Results  G-CSF upregulated the expression of leukemia cells miR-146a, down-regulated the mRNA expression and total protein expression of Smad4, and increased the proliferative activity of leukemia cells. Both mRNA expression and total protein expression of Smad4 of leukemia cells transfected by plasmids with overexpressed miR-146a decreased. The increase of expression of miR-146a and decrease of expression of Smad4 of transfected cells that were treated by G-CSF were more significant. Conclusion  G-CSF affects the expression of Smad4 via regulating the signal pathway of miR-146a/Smad4 and stimulates leukemia cells in quiescent state entering the proliferating phase.

Key words: granulocyte colony stimulating factor, miR-146/Smad4 expression, cell proliferation