Advertisement

Guanylate-binding protein 6 is a novel biomarker for tumorigenesis and prognosis in tongue squamous cell carcinoma

  • Pei-Feng LiuEmail author
  • Hung-Chih Chen
  • Chih-Wen Shu
  • Huei-Cin Sie
  • Cheng-Hsin Lee
  • Huei-Han Liou
  • Jiin-Tsuey Cheng
  • Kuo-Wang Tsai
  • Luo-Ping Ger
Original Article
  • 1 Downloads

Abstract

Objectives

Guanylate-binding protein 6 (GBP6) is a member of the guanylate-binding protein family, and its role in cancer has not yet been reported. We aimed to investigate the clinical significance of GBP6 in oral squamous cell carcinoma (OSCC).

Materials and methods

Next-generation sequencing was applied for analyzing differential gene expression profiling between corresponding tumor adjacent normal (CTAN) and tumor tissue from two paired OSCC patients. Real-time PCRs (RT-PCRs) were used to investigate the gene expression level of GBP6 of CTAN and tumor tissue samples from 14 TSCC patients. Immunohistochemistry was used to investigate the protein expression level of GBP6 in tumor tissues and paired CTAN tissues from 488 OSCC patients, including 183 buccal mucosa squamous cell carcinoma (BMSCC), 245 tongue squamous cell carcinoma (TSCC), and 60 lip squamous cell carcinoma (LSCC) patients.

Results

Compared with CTAN tissues of OSCC patients, GBP6 is identified as a downregulated gene using the NGS platform, which was confirmed in 14 OSCC patients by RT-PCR. Moreover, protein expression level of GBP6 in tumor tissues was lower than that in CTAN tissues and the low GBP6 expression was correlated with poor cell differentiation/lymph node metastasis in TSCC patients. In addition, TSCC patients with low expression levels of GBP6 had poor disease-specific survival rate.

Conclusion

The low expression of GBP6 was associated with tumorigenesis and poor prognosis in OSCC patients, especially in TSCC patients.

Clinical relevance

GBP6 may serve as a novel favorable diagnostic and prognostic biomarker in TSCC patients.

Keywords

Oral squamous cell carcinoma Guanylate-binding protein 6 Tumorigenesis Prognosis 

Abbreviations

BMSCC

Buccal mucosal squamous cell carcinoma

CTAN

Corresponding tumor adjacent normal

DFS

Disease-free survival

DSS

Disease-specific survival

IFN

Interferon

IHC

Immunohistochemistry

LCM

Laser capture microdissection

LSCC

Lip squamous cell carcinoma

OSCC

Oral squamous cell carcinoma

TSCC

Tongue squamous cell carcinoma

NGS

Next-generation sequencing

TMA

Tissue microarray

Notes

Funding information

This work was supported by grants from the Kaohsiung Veterans General Hospital (VGHKS107-174 and VGHKS108-178), the Ministry of Science and Technology (MOST 106-2320-B-075B-002-MY2 and MOST 108-2320-B-075B-003) and the Yen Tjing Ling Medical Foundation (CI-107-13).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Institutional Review Board (IRB) at the Kaohsiung Veterans General Hospital (KVGH, IRB number: VGHKS11-CT12-13) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from the parents of all individual participants included in the study.

Supplementary material

784_2019_3129_MOESM1_ESM.docx (459 kb)
ESM 1 (DOCX 458 kb)

References

  1. 1.
    Seta R, Mascitti M, Campagna R, Sartini D, Fumarola S, Santarelli A, Giuliani M, Cecati M, Muzio LL, Emanuelli M (2019) Overexpression of nicotinamide N-methyltransferase in HSC-2 OSCC cell line: effect on apoptosis and cell proliferation. Clin Oral Investig 23:829–838PubMedCrossRefGoogle Scholar
  2. 2.
    Maji S, Samal SK, Pattanaik L, Panda S, Quinn BA, Das SK, Sarkar D, Pellecchia M, Fisher PB, Dash R (2015) Mcl-1 is an important therapeutic target for oral squamous cell carcinomas. Oncotarget 6:16623–16637PubMedPubMedCentralCrossRefGoogle Scholar
  3. 3.
    Jadhav KB, Gupta N (2013) Clinicopathological prognostic implicators of oral squamous cell carcinoma: need to understand and revise. N Am J Med Sci 5:671–679PubMedPubMedCentralCrossRefGoogle Scholar
  4. 4.
    Curado MP, Hashibe M (2009) Recent changes in the epidemiology of head and neck cancer. Curr Opin Oncol 21:194–200PubMedCrossRefGoogle Scholar
  5. 5.
    Vestal DJ, Jeyaratnam JA (2011) The guanylate-binding proteins: emerging insights into the biochemical properties and functions of this family of large interferon-induced guanosine triphosphatase. J Interferon Cytokine Res 31:89–97PubMedPubMedCentralCrossRefGoogle Scholar
  6. 6.
    Ji C, Du S, Li P, Zhu Q, Yang X, Long C, Yu J, Shao F, Xiao J (2019) Structural mechanism for guanylate-binding proteins (GBPs) targeting by the Shigella E3 ligase IpaH9.8. PLoS Pathog 15:e1007876PubMedPubMedCentralCrossRefGoogle Scholar
  7. 7.
    Praefcke GJK (2018) Regulation of innate immune functions by guanylate-binding proteins. Int J Med Microbiol 308:237–245PubMedCrossRefGoogle Scholar
  8. 8.
    Britzen-Laurent N, Herrmann C, Naschberger E, Croner RS, Sturzl M (2016) Pathophysiological role of guanylate-binding proteins in gastrointestinal diseases. World J Gastroenterol 22:6434–6443PubMedPubMedCentralCrossRefGoogle Scholar
  9. 9.
    Budtz-Jorgensen E (1981) Oral mucosal lesions associated with the wearing of removable dentures. J Oral Pathol 10:65–80PubMedCrossRefGoogle Scholar
  10. 10.
    Vincent S, Marty L, Fort P (1993) S26 ribosomal protein RNA: an invariant control for gene regulation experiments in eucaryotic cells and tissues. Nucleic Acids Res 21:1498PubMedPubMedCentralCrossRefGoogle Scholar
  11. 11.
    Fu TY, Hsieh IC, Cheng JT, Tsai MH, Hou YY, Lee JH, Liou HH, Huang SF, Chen HC, Yen LM, Tseng HH, Ger LP (2015) Association of OCT4, SOX2, and NANOG expression with oral squamous cell carcinoma progression. J Oral Pathol MedGoogle Scholar
  12. 12.
    Chen HC, Tseng YK, Shu CW, Weng TJ, Liou HH, Yen LM, Hsieh IC, Wang CC, Wu PC, Shiue YL, Fu TY, Tsai KW, Ger LP, Liu PF (2019) Differential clinical significance of COL5A1 and COL5A2 in tongue squamous cell carcinoma. J Oral Pathol Med 48:468–476PubMedCrossRefGoogle Scholar
  13. 13.
    Piro AS, Hernandez D, Luoma S, Feeley EM, Finethy R, Yirga A, Frickel EM, Lesser CF, Coers J (2017) Detection of cytosolic Shigella flexneri via a C-terminal triple-arginine motif of GBP1 inhibits actin-based motility. MBio 8Google Scholar
  14. 14.
    Braun E, Hotter D, Koepke L, Zech F, Gross R, Sparrer KMJ, Muller JA, Pfaller CK, Heusinger E, Wombacher R, Sutter K, Dittmer U, Winkler M, Simmons G, Jakobsen MR, Conzelmann KK, Pohlmann S, Munch J, Fackler OT, Kirchhoff F, Sauter D (2019) Guanylate-binding proteins 2 and 5 exert broad antiviral activity by inhibiting furin-mediated processing of viral envelope proteins. Cell Rep 27:2092–2104 e10PubMedCrossRefGoogle Scholar
  15. 15.
    Britzen-Laurent N, Lipnik K, Ocker M, Naschberger E, Schellerer VS, Croner RS, Vieth M, Waldner M, Steinberg P, Hohenadl C, Sturzl M (2013) GBP-1 acts as a tumor suppressor in colorectal cancer cells. Carcinogenesis 34:153–162PubMedCrossRefGoogle Scholar
  16. 16.
    Friedman K, Brodsky AS, Lu S, Wood S, Gill AJ, Lombardo K, Yang D, Resnick MB (2016) Medullary carcinoma of the colon: a distinct morphology reveals a distinctive immunoregulatory microenvironment. Mod Pathol 29:528–541PubMedCrossRefGoogle Scholar
  17. 17.
    Godoy P, Cadenas C, Hellwig B, Marchan R, Stewart J, Reif R, Lohr M, Gehrmann M, Rahnenfuhrer J, Schmidt M, Hengstler JG (2014) Interferon-inducible guanylate binding protein (GBP2) is associated with better prognosis in breast cancer and indicates an efficient T cell response. Breast Cancer 21:491–499PubMedCrossRefGoogle Scholar
  18. 18.
    Lipnik K, Naschberger E, Gonin-Laurent N, Kodajova P, Petznek H, Rungaldier S, Astigiano S, Ferrini S, Sturzl M, Hohenadl C (2010) Interferon gamma-induced human guanylate binding protein 1 inhibits mammary tumor growth in mice. Mol Med 16:177–187PubMedPubMedCentralCrossRefGoogle Scholar
  19. 19.
    Yu CJ, Chang KP, Chang YJ, Hsu CW, Liang Y, Yu JS, Chi LM, Chang YS, Wu CC (2011) Identification of guanylate-binding protein 1 as a potential oral cancer marker involved in cell invasion using omics-based analysis. J Proteome Res 10:3778–3788PubMedCrossRefGoogle Scholar
  20. 20.
    Guimaraes DP, Oliveira IM, de Moraes E, Paiva GR, Souza DM, Barnas C, Olmedo DB, Pinto CE, Faria PA, De Moura Gallo CV, Small IA, Ferreira CG, Hainaut P (2009) Interferon-inducible guanylate binding protein (GBP)-2: a novel p53-regulated tumor marker in esophageal squamous cell carcinomas. Int J Cancer 124:272–279PubMedCrossRefGoogle Scholar
  21. 21.
    Fellenberg F, Hartmann TB, Dummer R, Usener D, Schadendorf D, Eichmuller S (2004) GBP-5 splicing variants: new guanylate-binding proteins with tumor-associated expression and antigenicity. J Invest Dermatol 122:1510–1517PubMedCrossRefGoogle Scholar
  22. 22.
    Persano L, Moserle L, Esposito G, Bronte V, Barbieri V, Iafrate M, Gardiman MP, Larghero P, Pfeffer U, Naschberger E, Sturzl M, Indraccolo S, Amadori A (2009) Interferon-alpha counteracts the angiogenic switch and reduces tumor cell proliferation in a spontaneous model of prostatic cancer. Carcinogenesis 30:851–860PubMedCrossRefGoogle Scholar
  23. 23.
    Guenzi E, Topolt K, Cornali E, Lubeseder-Martellato C, Jorg A, Matzen K, Zietz C, Kremmer E, Nappi F, Schwemmle M, Hohenadl C, Barillari G, Tschachler E, Monini P, Ensoli B, Sturzl M (2001) The helical domain of GBP-1 mediates the inhibition of endothelial cell proliferation by inflammatory cytokines. EMBO J 20:5568–5577PubMedPubMedCentralCrossRefGoogle Scholar
  24. 24.
    Walker K, Boyd NH, Anderson JC, Willey CD, Hjelmeland AB (2018) Kinomic profiling of glioblastoma cells reveals PLCG1 as a target in restricted glucose. Biomark Res 6:22PubMedPubMedCentralCrossRefGoogle Scholar
  25. 25.
    Mustafa DAM, Pedrosa R, Smid M, van der Weiden M, de Weerd V, Nigg AL, Berrevoets C, Zeneyedpour L, Priego N, Valiente M, Luider TM, Debets R, Martens JWM, Foekens JA, Sieuwerts AM, Kros JM (2018) T lymphocytes facilitate brain metastasis of breast cancer by inducing guanylate-binding protein 1 expression. Acta Neuropathol 135:581–599PubMedPubMedCentralCrossRefGoogle Scholar
  26. 26.
    Li L, Ma G, Jing C, Liu Z (2015) Guanylate-binding protein 1 (GBP1) promotes lymph node metastasis in human esophageal squamous cell carcinoma. Discov Med 20:369–378PubMedGoogle Scholar
  27. 27.
    Fukumoto M, Amanuma T, Kuwahara Y, Shimura T, Suzuki M, Mori S, Kumamoto H, Saito Y, Ohkubo Y, Duan Z, Sano K, Oguchi T, Kainuma K, Usami S, Kinoshita K, Lee I, Fukumoto M (2014) Guanine nucleotide-binding protein 1 is one of the key molecules contributing to cancer cell radioresistance. Cancer Sci 105:1351–1359PubMedPubMedCentralCrossRefGoogle Scholar
  28. 28.
    De Donato M, Mariani M, Petrella L, Martinelli E, Zannoni GF, Vellone V, Ferrandina G, Shahabi S, Scambia G, Ferlini C (2012) Class III beta-tubulin and the cytoskeletal gateway for drug resistance in ovarian cancer. J Cell Physiol 227:1034–1041PubMedCrossRefGoogle Scholar
  29. 29.
    Quintero M, Adamoski D, Reis LMD, Ascencao CFR, Oliveira KRS, Goncalves KA, Dias MM, Carazzolle MF, Dias SMG (2017) Guanylate-binding protein-1 is a potential new therapeutic target for triple-negative breast cancer. BMC Cancer 17:727PubMedPubMedCentralCrossRefGoogle Scholar
  30. 30.
    Li M, Mukasa A, Inda MM, Zhang J, Chin L, Cavenee W, Furnari F (2011) Guanylate binding protein 1 is a novel effector of EGFR-driven invasion in glioblastoma. J Exp Med 208:2657–2673PubMedPubMedCentralCrossRefGoogle Scholar
  31. 31.
    Zhang J, Zhang Y, Wu W, Wang F, Liu X, Shui G, Nie C (2017) Guanylate-binding protein 2 regulates Drp1-mediated mitochondrial fission to suppress breast cancer cell invasion. Cell Death Dis 8:e3151PubMedPubMedCentralCrossRefGoogle Scholar
  32. 32.
    Wang Q, Wang X, Liang Q, Wang S, Xiwen L, Pan F, Chen H, Li D (2018) Distinct prognostic value of mRNA expression of guanylate-binding protein genes in skin cutaneous melanoma. Oncol Lett 15:7914–7922PubMedPubMedCentralGoogle Scholar
  33. 33.
    Naschberger E, Croner RS, Merkel S, Dimmler A, Tripal P, Amann KU, Kremmer E, Brueckl WM, Papadopoulos T, Hohenadl C, Hohenberger W, Sturzl M (2008) Angiostatic immune reaction in colorectal carcinoma: impact on survival and perspectives for antiangiogenic therapy. Int J Cancer 123:2120–2129PubMedCrossRefGoogle Scholar
  34. 34.
    Anand R, Dhingra C, Prasad S, Menon I (2014) Betel nut chewing and its deleterious effects on oral cavity. J Cancer Res Ther 10:499–505PubMedCrossRefGoogle Scholar
  35. 35.
    Wu K, Wei J, Liu Z, Yu B, Yang X, Zhang C, Abdelrehem A, Zhang C, Li S (2019) Can pattern and depth of invasion predict lymph node relapse and prognosis in tongue squamous cell carcinoma. BMC Cancer 19:714PubMedPubMedCentralCrossRefGoogle Scholar
  36. 36.
    Han AY, Kuan EC, Mallen-St Clair J, Alonso JE, Arshi A, St John MA (2016) Epidemiology of squamous cell carcinoma of the lip in the United States: a population-based cohort analysis. JAMA Otolaryngol Head Neck Surg 142:1216–1223PubMedCrossRefGoogle Scholar
  37. 37.
    Gu T, Yu D, Fan Y, Wu Y, Yao YL, Xu L, Yao YG (2019) Molecular identification and antiviral function of the guanylate-binding protein (GBP) genes in the Chinese tree shrew (Tupaia belangeri chinesis). Dev Comp Immunol 96:27–36PubMedCrossRefGoogle Scholar
  38. 38.
    Capaldo CT, Beeman N, Hilgarth RS, Nava P, Louis NA, Naschberger E, Sturzl M, Parkos CA, Nusrat A (2012) IFN-gamma and TNF-alpha-induced GBP-1 inhibits epithelial cell proliferation through suppression of beta-catenin/TCF signaling. Mucosal Immunol 5:681–690PubMedPubMedCentralCrossRefGoogle Scholar
  39. 39.
    Unterer B, Wiesmann V, Gunasekaran M, Sticht H, Tenkerian C, Behrens J, Leone M, Engel FB, Britzen-Laurent N, Naschberger E, Wittenberg T, Sturzl M (2018) IFN-gamma-response mediator GBP-1 represses human cell proliferation by inhibiting the Hippo signaling transcription factor TEAD. Biochem J 475:2955–2967PubMedPubMedCentralCrossRefGoogle Scholar
  40. 40.
    Alspach E, Lussier DM, Schreiber RD (2019) Interferon gamma and its important roles in promoting and inhibiting spontaneous and therapeutic cancer immunity. Cold Spring Harb Perspect Biol 11Google Scholar
  41. 41.
    Whitmore SE, Lamont RJ (2014) Oral bacteria and cancer. PLoS Pathog 10:e1003933PubMedPubMedCentralCrossRefGoogle Scholar
  42. 42.
    Liu PF, Chang HW, Cheng JS, Lee HP, Yen CY, Tsai WL, Cheng JT, Li YJ, Huang WC, Lee CH, Ger LP, Shu CW (2018) Map1lc3b and Sqstm1 modulated autophagy for tumorigenesis and prognosis in certain subsites of oral squamous cell carcinoma. J Clin Med 7PubMedCentralCrossRefPubMedGoogle Scholar
  43. 43.
    Liu PF, Kang BH, Wu YM, Sun JH, Yen LM, Fu TY, Lin YC, Liou HH, Lin YS, Sie HC, Hsieh IC, Tseng YK, Shu CW, Hsieh YD, Ger LP (2017) Vimentin is a potential prognostic factor for tongue squamous cell carcinoma among five epithelial-mesenchymal transition-related proteins. PLoS One 12:e0178581PubMedPubMedCentralCrossRefGoogle Scholar
  44. 44.
    Fu TY, Wu CN, Sie HC, Cheng JT, Lin YS, Liou HH, Tseng YK, Shu CW, Tsai KW, Yen LM, Tseng HW, Tseng CJ, Ger LP, Liu PF (2016) Subsite-specific association of DEAD box RNA helicase DDX60 with the development and prognosis of oral squamous cell carcinoma. Oncotarget 7:85097–85108PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biomedical Science and Environmental BiologyKaohsiung Medical UniversityKaohsiungTaiwan
  2. 2.Institute of Biomedical SciencesNational Sun Yat-sen UniversityKaohsiungTaiwan
  3. 3.Department of StomatologyKaohsiung Veterans General HospitalKaohsiungTaiwan
  4. 4.Department of Dental TechnologyShu-Zen Junior College of Medicine and ManagementKaohsiungTaiwan
  5. 5.School of Medicine for International StudentsI-Shou UniversityKaohsiungTaiwan
  6. 6.Department of Pathology and Laboratory MedicineKaohsiung Veterans General HospitalKaohsiungTaiwan
  7. 7.Department of Medical Education and ResearchKaohsiung Veterans General HospitalKaohsiungTaiwan
  8. 8.Department of Biological SciencesNational Sun Yat-Sen UniversityKaohsiungTaiwan

Personalised recommendations