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Upregulation of CXCR4 is functionally crucial for maintenance of stemness in drug-resistant non-small cell lung cancer cells

Oncogene volume 32, pages 209–221 (2013)Cite this article

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Abstract

The hypothesis of cancer stem cells has been proposed to explain the therapeutic failure in a variety of cancers including lung cancers. Previously, we demonstrated acquisition of epithelial–mesenchymal transition, a feature highly reminiscent of cancer stem-like cells, in gefitinib-resistant A549 cells (A549/GR). Here, we show that A549/GR cells contain a high proportion of CXCR4+ cells that are responsible for having high potential of self-renewal activity in vitro and tumorigenicity in vivo. A549/GR cells exhibited strong sphere-forming activity and high CXCR4 expression and SDF-1α secretion compared with parent cells. Pharmacological inhibition (AMD3100) and/or siRNA transfection targeting CXCR4 significantly suppressed sphere-forming activity in A549 and A549/GR cells, and in various non-small cell lung cancer (NSCLC) cell lines. A549/GR cells showed enhanced Akt, mTOR and STAT3 (Y705) phosphorylation. Pharmacological inhibition of phosphatidyl inositol 3-kinase or transfection with wild-type PTEN suppressed phosphorylation of Akt, mTOR and STAT3 (Y705), sphere formation, and CXCR4 expression in A549/GR cells, whereas mutant PTEN enhanced these events. Inhibition of STAT3 by WP1066 or siSTAT3 significantly suppressed the sphere formation, but not CXCR4 expression, indicating that STAT3 is a downstream effector of CXCR4-mediated signaling. FACS-sorted CXCR4+ A549/GR cells formed many large spheres, had self-renewal capacity, demonstrated radiation resistance in vitro and exhibited stronger tumorigenic potential in vivo than CXCR4− cells. Lentiviral-transduction of CXCR4 enhanced sphere formation and tumorigenicity in H460 and A549 cells, whereas introduction of siCXCR4 suppressed these activities in A549/GR cells. Our data indicate that CXCR4+ NSCLC cells are strong candidates for tumorigenic stem-like cancer cells that maintain stemness through a CXCR4-medated STAT3 pathway and provide a potential therapeutic target for eliminating these malignant cells in NSCLC.

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Acknowledgements

This work was supported by the Mid-career Researcher Program through NRF grant (2009-0086438) and the Nuclear Research and Development Program of Korea Science and Engineering foundation funded by the Korean government (MEST) (2011-0030604).

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Author notes

  1. J C Lee and M-J Park: These two authors are contributed equally to this work

Authors and Affiliations

  1. Divisions of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences, Seoul, Korea

    M-J Jung, Y-M Kim, J E Jung, J-S Lee & M-J Park

  2. College of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea

    M-J Jung, Y-M Kim & Y-G Ko

  3. Department of Pulmonary and Critical Care Medicine, Asan Medical Center College of Medicine, University of Ulsan, Seoul, Korea

    J-K Rho

  4. Divisions of Radiation Effect, Korea Institute of Radiological and Medical Sciences, Seoul, Korea

    Y B Jin

  5. Department of Chemistry and Research Institute for Natural Sciences, Hanyang University, Seoul, Korea

    S-J Lee

  6. Department of Oncology, Asan Medical Center College of Medicine, University of Ulsan, Seoul, Korea

    J C Lee

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Jung, MJ., Rho, JK., Kim, YM. et al. Upregulation of CXCR4 is functionally crucial for maintenance of stemness in drug-resistant non-small cell lung cancer cells. Oncogene 32, 209–221 (2013). https://doi.org/10.1038/onc.2012.37

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