Chinese Medical Sciences Journal ›› 2013, Vol. 28 ›› Issue (2): 82-87.doi: 10.1016/S1001-9294(13)60027-9

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Lysine-specific Demethylase 1 Represses THP-1 Monocyte-to-macrophage Differentiation025B3;

Rui-feng Yang, Guo-wei Zhao, Shu-ting Liang, Hou-zao Chen, and De-pei Liu*   

  1. National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China
  • Revised:2012-10-15 Online:2013-06-26 Published:2013-06-26
  • Contact: 86-10-69156415,Fax:86-10-65105093,E-mail:liudp@pumc.edu.cn
  • Supported by:

    025B3;Supported by National Natural Science Foundation of China (31271227, 30721063, 81161120551) and National Basic Research Program of China (973 Program, 2011CB503902, 2011CB965203).

Abstract:

Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting were performed to analyze the expression of LSD1 and interleukin-6 (IL-6) in THP-1 monocytes and THP-1-derived macrophages. Chromatin immunoprecipitation (ChIP) assay was applied to detect the occupancy of LSD1 and H3K4 methylation at IL-6 promoter during THP-1 monocyte-to-macrophage differentiation. IL-6 mRNA level and H3K4 methylation at IL-6 promoter were analyzed using qRT-PCR and ChIP assay in LSD1-knockdown THP-1 cells treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) for 0, 4, 8, 12, and 24 hours. Fluorescence activated flow cytometry was performed to reveal the percentage of macrophages differentiated from THP-1 monocytes.

The expression of LSD1 reduced during THP-1 monocyte-to-macrophage differentiation (P<0.01). LSD1 occupancy decreased and H3K4 methylation increased at IL-6 promoter during the differentiation. With knockdown of LSD1, H3K4 methylation at IL-6 promoter was found increased after TPA treatment at different times points (all P<0.05, except 24 hours). The percentage of macrophages increased significantly in the THP-1 cells with LSD1 knockdown (P<0.05).

LSD1 is repressed during the monocyte-to-macrophage differentiation of THP-1 cells. Suppression of LSD1-mediated H3K4 demethylation may be required for THP-1 monocyte-to-macrophage differentiation.

Key words: lysine-specific demethylase 1, interleukin-6, H3K4 methylation, monocyte-to-macrophage differentiation

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