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EB-virus latent membrane protein 1 potentiates the stemness of nasopharyngeal carcinoma via preferential activation of PI3K/AKT pathway by a positive feedback loop

Oncogene volume 35, pages 3419–3431 (2016)Cite this article

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Abstract

Our previous study reported that Epstein–Barr virus(EBV)-encoded latent membrane protein 1 (LMP1) could induce development of CD44+/High stem-like cells in nasopharyngeal carcinoma (NPC). However, the molecular mechanisms that underlie modulation of cancer stem cells (CSCs) in NPC remain unclear. Here, we show that LMP1 induced CSC-like properties through promotion of the expression of epithelial–mesenchymal transition-like cellular markers and through alterations in differentiation markers. Furthermore, LMP1 activated and triggered phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathway, which subsequently stimulated expression of CSC markers, development of side population and tumor sphere formation. This suggests that PI3K/AKT pathway has an important role in the induction and maintenance of CSC properties in NPC. Similarly, PI3K/AKT pathway was also activated by phosphorylase in LMP1-induced CD44+/High cells. In addition, LMP1 greatly increased expression of miR-21 and downregulated expression of the miR-21 target, PTEN. Overexpression of miR-21 by transfection of miR-21 mimics into LMP1-transformed cells led to phosphorylase-mediated activation of the PI3K/AKT pathway and induction of CSCs. On the contrary, phosphorylation of the PI3K/AKT pathway and the expression of CSC were reversed by an miR-21 inhibitor. The specific inhibitor (Ly294002) of PI3K/AKT pathway significantly decreased expression of miR-21 and CSC markers and upregulated the expression of PTEN, which indicates that miR-21 and PTEN are the downstream effectors of PI3K/AKT and that expression of these two effectors are related to the development of NPC CSCs. Taken together, our novel findings indicate that LMP1, PI3K/AKT, miR-21 and PTEN constitute a positive feedback loop and have a key role in LMP1-induced CSCs in NPC.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (grant no. 81572901, 31170151, 81272340, 30770108 and 81030043), the Science and Technology Planning Project of Guangdong Province, China (grant no. 2014B020212017, 2014A020209024 and 2009A030331005), the Guangzhou Key Program of Science and Technology (grant no. 2012Y2-00026), the Shenzhen Basic Research Program of Science and Technology R&D (grant no. JC201105170709A and JCYJ20150330102720115), the Guangdong Natural Science Foundation (grant no. 2014A030310029) and the National High Technology Research and Development Program of China (863 Program; No. 2012AA02A501).

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Author notes

  1. C-F Yang, G-D Yang and T-J Huang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Experimental Research, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China

    C-F Yang, L Xu, C-N Qian & B-J Huang

  2. Department of Cancer Chemotherapy, Gaozhou People’s Hospital, Gaozhou, China

    C-F Yang, M-S Wang, M-D Cai, L Zhong & H-J Wei

  3. Department of Cancer Chemotherapy, Zengcheng People’s Hospital (BoJi-Affiliated Hospital of Sun Yat-Sen University), Zengcheng, China

    G-D Yang

  4. Department of Nuclear Medicine, The Second People’s Hospital of Shenzhen, Shenzhen, China

    T-J Huang

  5. Second College of Clinical Medicine, Southern Medical University, Guangzhou, China

    R Li

  6. School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou, China

    Q-Q Chu

  7. Department of Pharmacy, Sun Yat-Sen University Cancer Center, Guangzhou, China

    H-B Huang

  8. Department of Pathology, Saint Barnabas Medical Center, Livingston, NJ, USA

    J-L Huang

  9. Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, China

    C-N Qian

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Yang, CF., Yang, GD., Huang, TJ. et al. EB-virus latent membrane protein 1 potentiates the stemness of nasopharyngeal carcinoma via preferential activation of PI3K/AKT pathway by a positive feedback loop. Oncogene 35, 3419–3431 (2016). https://doi.org/10.1038/onc.2015.402

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