Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Stratifin regulates stabilization of receptor tyrosine kinases via interaction with ubiquitin-specific protease 8 in lung adenocarcinoma

Oncogene volume 37, pages 5387–5402 (2018)Cite this article

Subjects

Abstract

Previously we have reported that stratifin (SFN, 14-3-3 sigma) acts as a novel oncogene, accelerating the tumor initiation and progression of lung adenocarcinoma. Here, pull-down assay and LC-MS/MS analysis revealed that ubiquitin-specific protease 8 (USP8) specifically bound to SFN in lung adenocarcinoma cells. Both USP8 and SFN showed higher expression in human lung adenocarcinoma than in normal lung tissue, and USP8 expression was significantly correlated with SFN expression. Expression of SFN, but not of USP8, was associated with histological subtype, pathological stage, and poor prognosis. USP8 stabilizes receptor tyrosine kinases (RTKs) such as EGFR and MET by deubiquitination, contributing to the proliferative activity of many human cancers including non-small cell lung cancer. In vitro, USP8 binds to SFN and they co-localize at the early endosomes in lung adenocarcinoma cells. Moreover, USP8 or SFN knockdown leads to downregulation of tumor cellular proliferation and upregulation of apoptosis, p-EGFR or p-MET, which are related to the degradation pathway, and accumulation of ubiquitinated RTKs, leading to lysosomal degradation. Additionally, mutant USP8, which is unable to bind to SFN, reduces the expression of RTKs and p-STAT3. We also found that interaction with SFN is critical for USP8 to exert its autodeubiquitination function and avoid dephosphorylation by PP1. Our findings demonstrate that SFN enhances RTK stabilization through abnormal USP8 regulation in lung adenocarcinoma, suggesting that SFN could be a more suitable therapeutic target for lung adenocarcinoma than USP8.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin. 2016;66:7–30.

    Article  Google Scholar 

  2. Noguchi M, Morikawa A, Kawasaki M, Matsuno Y, Yamada T, Hirohashi S, et al. Small adenocarcinoma of the lung. Histologic characteristics and prognosis. Cancer. 1995;75:2844–52.

    Article  CAS  PubMed  Google Scholar 

  3. Kakinuma R, Noguchi M, Ashizawa K, Kuriyama K, Maeshima AM, Koizumi N, et al. Natural history of pulmonary subsolid nodules: a prospective multicenter study. J Thorac Oncol. 2016;11:1012–28.

    Article  PubMed  Google Scholar 

  4. Shiba-Ishii A, Kano J, Morishita Y, Sato Y, Minami Y, Noguchi M. High expression of stratifin is a universal abnormality during the course of malignant progression of early-stage lung adenocarcinoma. Int J Cancer. 2011;129:2445–53.

    Article  CAS  PubMed  Google Scholar 

  5. Shiba-Ishii A, Kim Y, Shiozawa T, Iyama S, Satomi K, Kano J, et al. Stratifin accelerates progression of lung adenocarcinoma at an early stage. Mol Cancer. 2015;14:142.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Aghazadeh Y, Papadopoulos V. The role of the 14-3-3 protein family in health, disease, and drug development. Drug Discov Today. 2016;21:278–87.

    Article  CAS  PubMed  Google Scholar 

  7. Yaffe MB, Rittinger K, Volinia S, Caron PR, Aitken A, Leffers H, et al. The structural basis for 14-3-3:phosphopeptide binding specificity. Cell. 1997;91:961–71.

    Article  CAS  PubMed  Google Scholar 

  8. Hermeking H. The 14-3-3 cancer connection. Nat Rev Cancer. 2003;3:931–43.

    Article  CAS  PubMed  Google Scholar 

  9. Smith AJ, Daut J, Schwappach B. Membrane proteins as 14-3-3 clients in functional regulation and intracellular transport. Physiology. 2011;26:181–91.

    Article  CAS  PubMed  Google Scholar 

  10. Dougherty MK, Morrison DK. Unlocking the code of 14-3-3. J Cell Sci. 2004;117:1875–84.

    Article  CAS  PubMed  Google Scholar 

  11. Fraile JM, Quesada V, Rodriguez D, Freije JM, Lopez-Otin C. Deubiquitinases in cancer: new functions and therapeutic options. Oncogene. 2012;31:2373–88.

    Article  CAS  PubMed  Google Scholar 

  12. Xia R, Jia H, Fan J, Liu Y, Jia J. USP8 promotes smoothened signaling by preventing its ubiquitination and changing its subcellular localization. PLoS Biol. 2012;10:e1001238.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Mukai A, Yamamoto-Hino M, Awano W, Watanabe W, Komada M, Goto S. Balanced ubiquitylation and deubiquitylation of Frizzled regulate cellular responsiveness to Wg/Wnt. EMBO J. 2010;29:2114–25.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Wu X, Yen L, Irwin L, Sweeney C, Carraway KL 3rd. Stabilization of the E3 ubiquitin ligase Nrdp1 by the deubiquitinating enzyme USP8. Mol Cell Biol. 2004;24:7748–57.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Mizuno E, Iura T, Mukai A, Yoshimori T, Kitamura N, Komada M. Regulation of epidermal growth factor receptor down-regulation by UBPY-mediated deubiquitination at endosomes. Mol Biol Cell. 2005;16:5163–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Meijer IM, van Leeuwen JE. ERBB2 is a target for USP8-mediated deubiquitination. Cell Signal. 2011;23:458–67.

    Article  CAS  PubMed  Google Scholar 

  17. Smith GA, Fearnley GW, Abdul-Zani I, Wheatcroft SB, Tomlinson DC, Harrison MA, et al. VEGFR2 trafficking, signaling and proteolysis is regulated by the ubiquitin isopeptidase USP8. Traffic. 2016;17:53–65.

    Article  CAS  PubMed  Google Scholar 

  18. Niendorf S, Oksche A, Kisser A, Lohler J, Prinz M, Schorle H, et al. Essential role of ubiquitin-specific protease 8 for receptor tyrosine kinase stability and endocytic trafficking in vivo. Mol Cell Biol. 2007;27:5029–39.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Takeuchi K, Ito F. Receptor tyrosine kinases and targeted cancer therapeutics. Biol Pharm Bull. 2011;34:1774–80.

    Article  CAS  PubMed  Google Scholar 

  20. Kim Y, Shiba-Ishii A, Nakagawa T, Husni RE, Sakashita S, Takeuchi T, et al. Ubiquitin-specific protease 8 is a novel prognostic marker in early-stage lung adenocarcinoma. Pathol Int. 2017;67:292–301.

    Article  CAS  PubMed  Google Scholar 

  21. Reincke M, Sbiera S, Hayakawa A, Theodoropoulou M, Osswald A, Beuschlein F, et al. Mutations in the deubiquitinase gene USP8 cause Cushing’s disease. Nat Genet. 2015;47:31–8.

    Article  CAS  PubMed  Google Scholar 

  22. Ma ZY, Song ZJ, Chen JH, Wang YF, Li SQ, Zhou LF, et al. Recurrent gain-of-function USP8 mutations in Cushing’s disease. Cell Res. 2015;25:306–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Ballif BA, Cao Z, Schwartz D, Carraway KL 3rd, Gygi SP. Identification of 14-3-3epsilon substrates from embryonic murine brain. J Proteome Res. 2006;5:2372–9.

    Article  CAS  PubMed  Google Scholar 

  24. Dufner A, Kisser A, Niendorf S, Basters A, Reissig S, Schonle A, et al. The ubiquitin-specific protease USP8 is critical for the development and homeostasis of T cells. Nat Immunol. 2015;16:950–60.

    Article  CAS  PubMed  Google Scholar 

  25. Mizuno E, Kitamura N, Komada M. 14-3-3-dependent inhibition of the deubiquitinating activity of UBPY and its cancellation in the M phase. Exp Cell Res. 2007;313:3624–34.

    Article  CAS  PubMed  Google Scholar 

  26. Benzinger A, Muster N, Koch HB, Yates JR 3rd, Hermeking H. Targeted proteomic analysis of 14-3-3 sigma, a p53 effector commonly silenced in cancer. Mol Cell Proteom. 2005;4:785–95.

    Article  CAS  Google Scholar 

  27. Shimada A, Kano J, Ishiyama T, Okubo C, Iijima T, Morishita Y, et al. Establishment of an immortalized cell line from a precancerous lesion of lung adenocarcinoma, and genes highly expressed in the early stages of lung adenocarcinoma development. Cancer Sci. 2005;96:668–75.

    Article  CAS  PubMed  Google Scholar 

  28. Mei Y, Hahn AA, Hu S, Yang X. The USP19 deubiquitinase regulates the stability of c-IAP1 and c-IAP2. J Biol Chem. 2011;286:35380–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Huang WG, Cheng AL, Chen ZC, Peng F, Zhang PF, Li MY, et al. Targeted proteomic analysis of 14-3-3sigma in nasopharyngeal carcinoma. Int J Biochem Cell Biol. 2010;42:137–47.

    Article  CAS  PubMed  Google Scholar 

  30. Byun S, Lee SY, Lee J, Jeong CH, Farrand L, Lim S, et al. USP8 is a novel target for overcoming gefitinib resistance in lung cancer. Clin Cancer Res. 2013;19:3894–904.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Li Z, Liu JY, Zhang JT. 14-3-3sigma, the double-edged sword of human cancers. Am J Transl Res. 2009;1:326–40.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Sbiera S, Deutschbein T, Weigand I, Reincke M, Fassnacht M, Allolio B. The new molecular landscape of Cushing’s disease. Trends Endocrinol Metab. 2015;26:573–83.

    Article  CAS  PubMed  Google Scholar 

  33. Panner A, Crane CA, Weng C, Feletti A, Fang S, Parsa AT, et al. Ubiquitin-specific protease 8 links the PTEN-Akt-AIP4 pathway to the control of FLIPS stability and TRAIL sensitivity in glioblastoma multiforme. Cancer Res. 2010;70:5046–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Wilker EW, Grant RA, Artim SC, Yaffe MB. A structural basis for 14-3-3sigma functional specificity. J Biol Chem. 2005;280:18891–8.

    Article  CAS  PubMed  Google Scholar 

  35. Shiba-Ishii A, Noguchi M. Aberrant stratifin overexpression is regulated by tumor-associated CpG demethylation in lung adenocarcinoma. Am J Pathol. 2012;180:1653–62.

    Article  CAS  PubMed  Google Scholar 

  36. Kim JO, Kim SR, Lim KH, Kim JH, Ajjappala B, Lee HJ, et al. Deubiquitinating enzyme USP37 regulating oncogenic function of 14-3-3gamma. Oncotarget. 2015;6:36551–76.

    PubMed  PubMed Central  Google Scholar 

  37. Alwan HA, van Leeuwen JE. UBPY-mediated epidermal growth factor receptor (EGFR) de-ubiquitination promotes EGFR degradation. J Biol Chem. 2007;282:1658–69.

    Article  CAS  PubMed  Google Scholar 

  38. Savio MG, Wollscheid N, Cavallaro E, Algisi V, Di Fiore PP, Sigismund S, et al. USP9X controls EGFR fate by deubiquitinating the endocytic adaptor Eps15. Curr Biol. 2016;26:173–83.

    Article  CAS  PubMed  Google Scholar 

  39. Pareja F, Ferraro DA, Rubin C, Cohen-Dvashi H, Zhang F, Aulmann S, et al. Deubiquitination of EGFR by Cezanne-1 contributes to cancer progression. Oncogene. 2012;31:4599–608.

    Article  CAS  PubMed  Google Scholar 

  40. Clague MJ, Urbe S. Endocytosis: the DUB version. Trends Cell Biol. 2006;16:551–9.

    Article  CAS  PubMed  Google Scholar 

  41. Row PE, Prior IA, McCullough J, Clague MJ, Urbe S. The ubiquitin isopeptidase UBPY regulates endosomal ubiquitin dynamics and is essential for receptor down-regulation. J Biol Chem. 2006;281:12618–24.

    Article  CAS  PubMed  Google Scholar 

  42. Komada M. Controlling receptor downregulation by ubiquitination and deubiquitination. Curr Drug Discov Technol. 2008;5:78–84.

    Article  CAS  PubMed  Google Scholar 

  43. Ledda F, Paratcha G. Negative regulation of receptor tyrosine kinase (RTK) signaling: a developing field. Biomark Insights. 2007;2:45–58.

    Article  PubMed  PubMed Central  Google Scholar 

  44. Guo L, Kozlosky CJ, Ericsson LH, Daniel TO, Cerretti DP, Johnson RS. Studies of ligand-induced site-specific phosphorylation of epidermal growth factor receptor. J Am Soc Mass Spectrom. 2003;14:1022–31.

    Article  CAS  PubMed  Google Scholar 

  45. Perez-Rivas LG, Theodoropoulou M, Ferrau F, Nusser C, Kawaguchi K, Stratakis CA, et al. The gene of the ubiquitin-specific protease 8 is frequently mutated in adenomas causing Cushing’s disease. J Clin Endocrinol Metab. 2015;100:E997–1004.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Meijer IM, Kerperien J, Sotoca AM, van Zoelen EJ, van Leeuwen JE. The Usp8 deubiquitination enzyme is post-translationally modified by tyrosine and serine phosphorylation. Cell Signal. 2013;25:919–30.

    Article  CAS  PubMed  Google Scholar 

  47. Devarakonda S, Morgensztern D, Govindan R. Genomic alterations in lung adenocarcinoma. Lancet Oncol. 2015;16:e342–51.

    Article  CAS  PubMed  Google Scholar 

  48. Bansal P, Osman D, Gan GN, Simon GR, Boumber Y. Recent advances in targetable therapeutics in metastatic non-squamous NSCLC. Front Oncol. 2016;6:112.

    PubMed  PubMed Central  Google Scholar 

  49. Naviglio S, Mattecucci C, Matoskova B, Nagase T, Nomura N, Di Fiore PP, et al. UBPY: a growth-regulated human ubiquitin isopeptidase. EMBO J. 1998;17:3241–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Sato T, Shiba-Ishii A, Kim Y, Dai T, Husni RE, Hong J, et al. miR-3941: a novel microRNA that controls IGBP1 expression and is associated with malignant progression of lung adenocarcinoma. Cancer Sci. 2017;108:536–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Travis WD, Brambilla E, Noguchi M, Nicholson AG, Geisinger KR, Yatabe Y, et al. International association for the study of lung cancer/american thoracic society/european respiratory society international multidisciplinary classification of lung adenocarcinoma. J Thorac Oncol. 2011;6:244–85.

    Article  PubMed  PubMed Central  Google Scholar 

  52. Travis WD, Brambilla E, Nicholson AG, Yatabe Y, Austin JH, Beasley MB, et al. The2015 world health organization classification of lung tumors: impact of genetic, clinical and radiologic advances since the 2004 classification. J Thorac Oncol. 2015;10:1243–60.

    Article  Google Scholar 

Download references

Acknowledgements

We express our appreciation to Professor Yasunori Kanaho and Dr. Yuji Funakoshi for research support and kindly providing the plasmids and reagents. We also thank Professor Mitsuyasu Kato and Dr. Hiroyuki Suzuki (Faculty of Medicine, University of Tsukuba) for research support and Professor Flaminia Miyamasu (Medical English Communications Center, University of Tsukuba) for critical review of this manuscript.

Author information

Author notes

  1. These authors contributed equally: Yunjung Kim, Aya Shiba-Ishii

Authors and Affiliations

  1. Department of Pathology, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan

    Yunjung Kim, Aya Shiba-Ishii, Noriyuki Nakano, Ryota Matsuoka, Shingo Sakashita & Masayuki Noguchi

  2. Doctoral Program in Biomedical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Ibaraki, Japan

    Tomoki Nakagawa

  3. Translational Research Center, Fukushima Medical University, Fukushima, Japan

    Shun-ichiro Iemura

  4. Molecular Profiling Research Center for Drug Discovery, National Institutes of Advanced Industrial Science and Technology, Tokyo, Japan

    Tohru Natsume

  5. Ph.D. Program in Human Biology, School of Integrative and Global Majors, University of Tsukuba, Ibaraki, Japan

    SangJoon Lee

  6. Department of Infection Biology, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan

    Atsushi Kawaguchi

  7. Department of Thoracic Surgery, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan

    Yukio Sato

Authors
  1. Yunjung Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Aya Shiba-Ishii
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tomoki Nakagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shun-ichiro Iemura
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tohru Natsume
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Noriyuki Nakano
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ryota Matsuoka
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Shingo Sakashita
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. SangJoon Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Atsushi Kawaguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Yukio Sato
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Masayuki Noguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Aya Shiba-Ishii or Masayuki Noguchi.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kim, Y., Shiba-Ishii, A., Nakagawa, T. et al. Stratifin regulates stabilization of receptor tyrosine kinases via interaction with ubiquitin-specific protease 8 in lung adenocarcinoma. Oncogene 37, 5387–5402 (2018). https://doi.org/10.1038/s41388-018-0342-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-018-0342-9

This article is cited by

Search

Advanced search

Quick links