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Regulatory mechanism and functional analysis of S100A9 in acute promyelocytic leukemia cells

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Abstract

S100A9, a calcium-binding protein, participates in the inflammatory process and development of various tumors, thus attracting much attention in the field of cancer biology. This study aimed to investigate the regulatory mechanism of S100A9 and its function involvement in APL. We used real-time quantitative PCR to determine whether PML/RARα affects the expression of S100A9 in NB4 and PR9 cells upon ATRA treatment. ChIP-based PCR and dual-luciferase reporter assay system were used to detect how PML/RARα and PU.1 regulate S100A9 promoter activity. CCK-8 assay and flow cytometry were employed to observe the viability and apoptosis of NB4 cells when S100A9 was overexpressed. Results showed that S100A9 was an ATRA-responsive gene, and PML/RARα was necessary for the ATRA-induced expression of S100A9 in APL cells. In addition, PU.1 could bind to the promoter of S100A9, especially when treated with ATRA in NB4 cells, and promote its activity. More importantly, overexpression of S100A9 induced the apoptosis of NB4 cells and inhibited cell growth. Collectively, our data indicated that PML/RARα and PU.1 were necessary for the ATRA-induced expression of S100A9 in APL cells. Furthermore, S100A9 promoted apoptosis in APL cells and affected cell growth.

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Acknowledgements

This work was supported in part by the National Natural Science Foundation of China (No. 81370655), and the grants from Ministry of Science and Technology of China (Nos. 2013CB966802 and 2012AA02A505). We also greatly appreciated institutional supports from Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, China.

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Authors and Affiliations

  1. State Key Laboratory of Medical Genomics, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China

    Yonglan Zhu, Fang Zhang, Wanglong Deng, Huiyong Fan, Haiwei Wang & Ji Zhang

  2. Medical Institute of Health Sciences, Chinese Academy of Sciences, Shanghai, 200025, China

    Shanzhen Zhang, Haiwei Wang & Ji Zhang

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  1. Yonglan Zhu
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  2. Fang Zhang
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  3. Shanzhen Zhang
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  7. Ji Zhang
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Correspondence to Ji Zhang.

Electronic supplementary material

Fig.S1

Arsenic trioxide had slight effect on the expression level of S100A9. The expression levels of S100A9 and PU.1 were detected at 24 h and 48 h treated with 1μmol/L arsenic trioxide in NB4 cells. GAPDH was used for normalization. The expression levels of S100A9 were examined at 24 h and 48 h treated with 1μmol/L ATRA or arsenic trioxide in NB4 cells. GAPDH was used for normalization. The data represent the mean of three replicates ± S.D.

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Zhu, Y., Zhang, F., Zhang, S. et al. Regulatory mechanism and functional analysis of S100A9 in acute promyelocytic leukemia cells. Front. Med. 11, 87–96 (2017). https://doi.org/10.1007/s11684-016-0469-4

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