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Sirtuin2 enhances the tumoricidal function of liver natural killer cells in a mouse hepatocellular carcinoma model

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Abstract

Hepatocellular carcinoma (HCC) is the third most lethal cancer in the world. Natural killer (NK) cell-mediated immunity is crucial for tumor surveillance and therapy. Characterization of the regulatory mechanisms of NK cell function is important for developing novel immunotherapies against HCC. In this study, we used a chemical-induced mouse HCC model to identify the upregulation of Sirtuin2 (SIRT2) in liver NK cells. In particular, SIRT2 was predominantly expressed in liver CD94+ NK cells. The HCC liver microenvironment induced SIRT2 expression in NK cells. In addition, overexpression of exogenous SIRT2 significantly upregulated the production of cytokines and cytotoxic mediators in activated NK cells. Consistently, SIRT2-overexpressing NK cells showed a stronger tumoricidal effect on hepatoma cells. Moreover, SIRT2 remarkably promoted the phosphorylation of Extracellular-signal-regulated kinase 1/2 (Erk1/2) and p38 Mitogen-activated protein kinases (MAPK) in activated NK cells. SIRT2 knockdown in liver CD94+ NK cells impaired their cytotoxic effect on hepatoma cells. Our study indicates that SIRT2 enhances the tumoricidal activity of liver NK cells in HCC.

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Abbreviations

ACK:

Ammonium–chloride–potassium

APC:

Allophycocyanin

Erk:

Extracellular-signal-regulated kinase

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

GFP:

Green fluorescent protein

HCC:

Hepatocellular carcinoma

JNK:

Jun N-terminal kinase

MAPK:

Mitogen-activated protein kinases

MOI:

Multiplicity of infection

NAD:

Nicotinamide adenine dinucleotide

NK cells:

Natural killer cells

NKG2:

Natural killer group 2

PBS:

Phosphate-buffered saline

RIPA:

Radioimmunoprecipitation assay

SIRT:

Sirtuin

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Funding

This study was supported by the National Natural Science Foundation of China (Grant No. 81470823) and the Natural Science Foundation of Hubei Province (Grant No. 2013CFB304).

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Author notes

  1. Ming Chen and Min Xu contributed equally to this work.

Authors and Affiliations

  1. Department of Blood Transfusion, Zhongnan Hospital of Wuhan University, Wuhan, China

    Ming Chen

  2. Department of Hematology and Oncology, Wuhan Children’s Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

    Min Xu

  3. Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan, China

    Chengliang Zhu

  4. Hubei Clinical Center and Key Laboratory for Intestinal and Colorectal Disease, Department of Gastroenterology, Zhongnan Hospital of Wuhan University, 169 East Lake Road, Wuchang District, Wuhan City, Hubei Province, China

    Hongling Wang, Qiu Zhao & Feng Zhou

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  1. Ming Chen
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  4. Hongling Wang
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Contributions

MC conducted the animal model, cell sorting, quantitative RT-PCR, adoptive transfer, and in vitro culture. MX prepared the lentiviruses and performed the lentiviral transduction. CZ conducted the immunoblotting analysis. HW did the intracellular staining and flow cytometry analysis. QZ conducted the statistical analysis. FZ designed the experiments, analyzed the data and composed the manuscript.

Corresponding author

Correspondence to Feng Zhou.

Ethics declarations

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

The animal study was approved by the Wuhan University Animal Care and Use Committee (animal research approval number: 11400700245566). The animal experiments were performed following the Wuhan University Animal Use Guidelines.

Informed consent

Not applicable because no human subjects were included.

Animal source

All mice were obtained from SBS Genetech Co, Ltd.

Cell line authentication

Not applicable because primary NK cells were used. The mouse hepatoma cell line Hepa1-6RAE1 was a gift from another lab (from Dr. Kai Dai, Wuhan University) so no authentication information was gained. The HEK293T cell line was purchased from Procell Life Science & Technology Co, Ltd. (Wuhan, China) and was not authenticated, because this cell line was only used for lentivirus packaging in the current study.

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Chen, M., Xu, M., Zhu, C. et al. Sirtuin2 enhances the tumoricidal function of liver natural killer cells in a mouse hepatocellular carcinoma model. Cancer Immunol Immunother 68, 961–971 (2019). https://doi.org/10.1007/s00262-019-02337-5

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  • DOI: https://doi.org/10.1007/s00262-019-02337-5

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