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NANOG modulates stemness in human colorectal cancer

Oncogene volume 32, pages 4397–4405 (2013)Cite this article

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Abstract

NANOG is a stem cell transcription factor that is essential for embryonic development, reprogramming normal adult cells and malignant transformation and progression. The nearly identical retrogene NANOGP8 is expressed in multiple cancers, but generally not in normal tissues and its function is not well defined. Our postulate is that NANOGP8 directly modulates the stemness of individual human colorectal carcinoma (CRC) cells. Stemness was measured in vitro as the spherogenicity of single CRC cells in serum-free medium and the size of the side population (SP) and in vivo as tumorigenicity and experimental metastatic potential in NOD/SCID mice. We found that 80% of clinical liver metastases express a NANOG with 75% of the positive metastases containing NANOGP8 transcripts. In all, 3–62% of single cells within six CRC lines form spheroids in serum-free medium in suspension. NANOGP8 is translated into protein. The relative expression of a NANOG gene increased 8- to 122-fold during spheroid formation, more than the increase in OCT4 or SOX2 transcripts with NANOGP8 the more prevalent family member. Short hairpin RNA (shRNA) to NANOG not only inhibits spherogenicity but also reduces expression of OCT4 and SOX2, the size of the SP and tumor growth in vivo. Inhibition of NANOG gene expression is associated with inhibition of proliferation and decreased phosphorylation of G2-related cell-cycle proteins. Overexpression of NANOGP8 rescues single-cell spherogenicity when NANOG gene expression is inhibited and increases the SP in CRC. Thus, NANOGP8 can substitute for NANOG in directly promoting stemness in CRC.

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Acknowledgements

We acknowledge the valuable advice and support of Drs Elizabeth Conner and Valentina Factor. Also, we gratefully acknowledge the support provided by the Center for Cancer Research of the NCI for Project ZIA BC 011199 and by the Department of Defense for Grant Number W81XWH-11-1-0327. The opinions expressed in this manuscript are those of the authors and do not necessarily represent those of the National Cancer Institute, the National Institutes of Health, the Department of Health and Human Services or the Department of the Army.

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  1. J Zhang and L A Espinoza: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Experimental Carcinogenesis, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    J Zhang, L A Espinoza, S S Thorgeirsson & J M Jessup

  2. Laboratory of Human Toxicology and Pharmacology, Applied/Developmental Research Support Directorate, Frederick National Laboratory for Cancer Research, SAIC-Frederick, Inc., Frederick, MD, USA

    R J Kinders, S M Lawrence & T D Pfister

  3. Laboratory of Proteomics and Analytical Technologies, Frederick National Laboratory for Cancer Research, SAIC-Frederick, Frederick, MD, USA

    M Zhou & T D Veenstra

  4. Division of Cancer Treatment and Diagnosis, Cancer Diagnosis Program, Rockville, MD, USA

    J M Jessup

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  1. J Zhang
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Zhang, J., Espinoza, L., Kinders, R. et al. NANOG modulates stemness in human colorectal cancer. Oncogene 32, 4397–4405 (2013). https://doi.org/10.1038/onc.2012.461

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