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Overexpression of S100A4 Predicts Migration, Invasion, and Poor Prognosis of Hypopharyngeal Squamous Cell Carcinoma

  • Jianing Xu
  • Neil Gross
  • Yuanwei Zang
  • Shengda Cao
  • Feilong Yang
  • Zheng Yang
  • Wenbin YuEmail author
  • Dapeng Lei
  • Xinliang PanEmail author
Original Research Article
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Abstract

Introduction

Hypopharyngeal squamous cell carcinoma (HSCC) is among the most lethal tumors encountered in the head and neck and frequently involves regional metastasis. However, the mechanism underlying the aggressiveness of HSCC remains elusive. S100A4 is a well-established metastasis-promoting regulator in a variety of malignancies, but its role in HSCC has not yet been identified.

Objectives

Our objectives were to explore the expression levels of S100A4 in HSCC tumors and its association with clinicopathological parameters and the clinical prognosis of HSCC and to confirm its role in the metastatic process of the HSCC FaDu cell line in vitro.

Methods

We assessed the expression levels of S100A4 with immunohistochemistry (IHC) in HSCC tumors (n = 71) and adjacent normal tissues (n = 44). In vitro experiments were performed to explore the impact of S100A4 knockdown on biological phenotypes of human HSCC FaDu cell line, including migration, invasion, proliferation, apoptosis, and cell cycle.

Results

The expression of S100A4 was elevated in HSCC tumors compared with adjacent normal tissues and positively correlated with cervical lymph node metastasis in this HSCC patient cohort. In vitro experiments showed that S100A4 knockdown significantly impaired migration and invasion and increased the proportion of cells in G0/G1 phase with no change in proliferation or apoptosis in FaDu cells. Additionally, nuclear S100A4 expression proved to be an independent prognostic indicator in patients with HSCC.

Conclusion

This study demonstrated for the first time that S100A4 expression is upregulated in HSCC tumors and that this upregulation is positively correlated with cervical lymph node metastasis of this malignancy. The metastasis-promoting role of S100A4 was further validated in the HSCC FaDu cell line, indicating that S100A4 is a potential therapeutic target for HSCC. Furthermore, this study suggests that nuclear S100A4 expression could be considered a prognostic biomarker for HSCC.

Notes

Compliance with Ethical Standards

Conflicts of interest

J Xu, N. Gross, Y. Zang, S. Cao, F. Yang, Z. Yang, W. Yu, D. Lei, X. Pan have no conflicts of interest that are directly relevant to the content of this article.

Ethics approval

This study protocol was approved by the Ethics Committee of Qilu Hospital of Shandong University.

Informed consent

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

Funding

This work was supported by the Taishan Scholars Program (No. tshw20130950), Shandong Province, and the Department of Science & Technology of Shandong Province (Nos. ZR2013HM107, ZR2014HM005, 2015GSF118014, 2015GSF118030 and 2018GSF18166), and Science Foundation of Qilu Hospital of Shandong University; and the Fundamental Research Funds of Shandong University (No. 2014QLKY05).

References

  1. 1.
    Gillison ML, Koch WM, Capone RB, Spafford M, Westra WH, Wu L, et al. Evidence for a causal association between human papillomavirus and a subset of head and neck cancers. J Natl Cancer Inst. 2000;92(9):709–20.CrossRefGoogle Scholar
  2. 2.
    Yan M, Xu Q, Zhang P, Zhou XJ, Zhang ZY, Chen WT. Correlation of NF-kappaB signal pathway with tumor metastasis of human head and neck squamous cell carcinoma. BMC Cancer. 2010;10:437.CrossRefGoogle Scholar
  3. 3.
    Bornstein S, White R, Malkoski S, Oka M, Han G, Cleaver T, et al. Smad4 loss in mice causes spontaneous head and neck cancer with increased genomic instability and inflammation. J Clin Investig. 2009;119(11):3408–19.Google Scholar
  4. 4.
    Warnakulasuriya S. Global epidemiology of oral and oropharyngeal cancer. Oral Oncol. 2009;45(4–5):309–16.CrossRefGoogle Scholar
  5. 5.
    Gourin CG, Johnson JT. A contemporary review of indications for primary surgical care of patients with squamous cell carcinoma of the head and neck. Laryngoscope. 2009;119(11):2124–34.CrossRefGoogle Scholar
  6. 6.
    Kotwall C, Sako K, Razack MS, Rao U, Bakamjian V, Shedd DP. Metastatic patterns in squamous cell cancer of the head and neck. Am J Surg. 1987;154(4):439–42.CrossRefGoogle Scholar
  7. 7.
    Takes RP, Strojan P, Silver CE, Bradley PJ, Haigentz M Jr, Wolf GT, et al. Current trends in initial management of hypopharyngeal cancer: the declining use of open surgery. Head Neck. 2012;34(2):270–81.CrossRefGoogle Scholar
  8. 8.
    Ausoni S, Boscolo-Rizzo P, Singh B, Da Mosto MC, Spinato G, Tirelli G, et al. Targeting cellular and molecular drivers of head and neck squamous cell carcinoma: current options and emerging perspectives. Cancer Metastasis Rev. 2016;35(3):413–26.CrossRefGoogle Scholar
  9. 9.
    Jin K, Li T, van Dam H, Zhou F, Zhang L. Molecular insights into tumour metastasis: tracing the dominant events. J Pathol. 2017;241(5):567–77.CrossRefGoogle Scholar
  10. 10.
    Boye K, Maelandsmo GM. S100A4 and metastasis: a small actor playing many roles. Am J Pathol. 2010;176(2):528–35.CrossRefGoogle Scholar
  11. 11.
    Grigorian M, Ambartsumian N, Lykkesfeldt AE, Bastholm L, Elling F, Georgiev G, et al. Effect of mts1 (S100A4) expression on the progression of human breast cancer cells. Int J Cancer. 1996;67(6):831–41.CrossRefGoogle Scholar
  12. 12.
    Maelandsmo GM, Hovig E, Skrede M, Engebraaten O, Florenes VA, Myklebost O, et al. Reversal of the in vivo metastatic phenotype of human tumor cells by an anti-CAPL (mts1) ribozyme. Cancer Res. 1996;56(23):5490–8.Google Scholar
  13. 13.
    Takenaga K, Nakamura Y, Sakiyama S. Expression of antisense RNA to S100A4 gene encoding an S100-related calcium-binding protein suppresses metastatic potential of high-metastatic Lewis lung carcinoma cells. Oncogene. 1997;14(3):331–7.CrossRefGoogle Scholar
  14. 14.
    Levett D, Flecknell PA, Rudland PS, Barraclough R, Neal DE, Mellon JK, et al. Transfection of S100A4 produces metastatic variants of an orthotopic model of bladder cancer. Am J Pathol. 2002;160(2):693–700.CrossRefGoogle Scholar
  15. 15.
    Ambartsumian NS, Grigorian MS, Larsen IF, Karlstrom O, Sidenius N, Rygaard J, et al. Metastasis of mammary carcinomas in GRS/A hybrid mice transgenic for the mts1 gene. Oncogene. 1996;13(8):1621–30.Google Scholar
  16. 16.
    Davies MP, Rudland PS, Robertson L, Parry EW, Jolicoeur P, Barraclough R. Expression of the calcium-binding protein S100A4 (p9Ka) in MMTV-neu transgenic mice induces metastasis of mammary tumours. Oncogene. 1996;13(8):1631–7.Google Scholar
  17. 17.
    Grum-Schwensen B, Klingelhofer J, Grigorian M, Almholt K, Nielsen BS, Lukanidin E, et al. Lung metastasis fails in MMTV-PyMT oncomice lacking S100A4 due to a T-cell deficiency in primary tumors. Cancer Res. 2010;70(3):936–47.CrossRefGoogle Scholar
  18. 18.
    Atlasi Y, Noori R, Marolin I, Franken P, Brandao J, Biermann K, et al. The role of S100a4 (Mts1) in Apc- and Smad4-driven tumour onset and progression. Eur J Cancer. 2016;68:114–24.CrossRefGoogle Scholar
  19. 19.
    Keirsebilck A, Bonne S, Bruyneel E, Vermassen P, Lukanidin E, Mareel M, et al. E-cadherin and metastasin (mts-1/S100A4) expression levels are inversely regulated in two tumor cell families. Cancer Res. 1998;58(20):4587–91.Google Scholar
  20. 20.
    Stein U, Arlt F, Walther W, Smith J, Waldman T, Harris ED, et al. The metastasis-associated gene S100A4 is a novel target of beta-catenin/T-cell factor signaling in colon cancer. Gastroenterology. 2006;131(5):1486–500.CrossRefGoogle Scholar
  21. 21.
    Siddique HR, Adhami VM, Parray A, Johnson JJ, Siddiqui IA, Shekhani MT, et al. The S100A4 oncoprotein promotes prostate tumorigenesis in a transgenic mouse model: regulating NFkappaB through the RAGE Receptor. Genes Cancer. 2013;4(5–6):224–34.CrossRefGoogle Scholar
  22. 22.
    Hernan R, Fasheh R, Calabrese C, Frank AJ, Maclean KH, Allard D, et al. ERBB2 up-regulates S100A4 and several other prometastatic genes in medulloblastoma. Cancer Res. 2003;63(1):140–8.Google Scholar
  23. 23.
    Ambartsumian N, Klingelhofer J, Grigorian M, Christensen C, Kriajevska M, Tulchinsky E, et al. The metastasis-associated Mts1(S100A4) protein could act as an angiogenic factor. Oncogene. 2001;20(34):4685–95.CrossRefGoogle Scholar
  24. 24.
    Schmidt-Hansen B, Ornas D, Grigorian M, Klingelhofer J, Tulchinsky E, Lukanidin E, et al. Extracellular S100A4(mts1) stimulates invasive growth of mouse endothelial cells and modulates MMP-13 matrix metalloproteinase activity. Oncogene. 2004;23(32):5487–95.CrossRefGoogle Scholar
  25. 25.
    Zhang HY, Zheng XZ, Wang XH, Xuan XY, Wang F, Li SS. S100A4 mediated cell invasion and metastasis of esophageal squamous cell carcinoma via the regulation of MMP-2 and E-cadherin activity. Mol Biol Rep. 2012;39(1):199–208.CrossRefGoogle Scholar
  26. 26.
    Schmidt-Hansen B, Klingelhofer J, Grum-Schwensen B, Christensen A, Andresen S, Kruse C, et al. Functional significance of metastasis-inducing S100A4(Mts1) in tumor-stroma interplay. J Biol Chem. 2004;279(23):24498–504.CrossRefGoogle Scholar
  27. 27.
    Nasser MW, Elbaz M, Ahirwar DK, Ganju RK. Conditioning solid tumor microenvironment through inflammatory chemokines and S100 family proteins. Cancer Lett. 2015;365(1):11–22.CrossRefGoogle Scholar
  28. 28.
    Grum-Schwensen B, Klingelhofer J, Berg CH, El-Naaman C, Grigorian M, Lukanidin E, et al. Suppression of tumor development and metastasis formation in mice lacking the S100A4(mts1) gene. Cancer Res. 2005;65(9):3772–80.CrossRefGoogle Scholar
  29. 29.
    O’Connell JT, Sugimoto H, Cooke VG, MacDonald BA, Mehta AI, LeBleu VS, et al. VEGF-A and Tenascin-C produced by S100A4 + stromal cells are important for metastatic colonization. Proc Natl Acad Sci USA. 2011;108(38):16002–7.CrossRefGoogle Scholar
  30. 30.
    Fei F, Qu J, Zhang M, Li Y, Zhang S. S100A4 in cancer progression and metastasis: a systematic review. Oncotarget. 2017;8(42):73219–39.CrossRefGoogle Scholar
  31. 31.
    Boye K, Nesland JM, Sandstad B, Maelandsmo GM, Flatmark K. Nuclear S100A4 is a novel prognostic marker in colorectal cancer. Eur J Cancer. 2010;46(16):2919–25.CrossRefGoogle Scholar
  32. 32.
    Boye K, Jacob H, Frikstad KA, Nesland JM, Maelandsmo GM, Dahl O, et al. Prognostic significance of S100A4 expression in stage II and III colorectal cancer: results from a population-based series and a randomized phase III study on adjuvant chemotherapy. Cancer Med. 2016;5(8):1840–9.CrossRefGoogle Scholar
  33. 33.
    Egeland EV, Boye K, Park D, Synnestvedt M, Sauer T, Oslo Breast Cancer C, et al. Prognostic significance of S100A4-expression and subcellular localization in early-stage breast cancer. Breast Cancer Res Treat. 2017;162(1):127-37.Google Scholar
  34. 34.
    Platt-Higgins AM, Renshaw CA, West CR, Winstanley JH, De Silva Rudland S, Barraclough R, et al. Comparison of the metastasis-inducing protein S100A4 (p9 ka) with other prognostic markers in human breast cancer. Int J Cancer. 2000;89(2):198–208.CrossRefGoogle Scholar
  35. 35.
    Davies BR, O’Donnell M, Durkan GC, Rudland PS, Barraclough R, Neal DE, et al. Expression of S100A4 protein is associated with metastasis and reduced survival in human bladder cancer. J Pathol. 2002;196(3):292–9.CrossRefGoogle Scholar
  36. 36.
    Lo JF, Yu CC, Chiou SH, Huang CY, Jan CI, Lin SC, et al. The epithelial-mesenchymal transition mediator S100A4 maintains cancer-initiating cells in head and neck cancers. Cancer Res. 2011;71(5):1912–23.CrossRefGoogle Scholar
  37. 37.
    Misaka T, Murakawa T, Nishida K, Omori Y, Taneike M, Omiya S, et al. FKBP8 protects the heart from hemodynamic stress by preventing the accumulation of misfolded proteins and endoplasmic reticulum-associated apoptosis in mice. J Mol Cell Cardiol. 2018;114:93–104.CrossRefGoogle Scholar
  38. 38.
    Rangan SR. A new human cell line (FaDu) from a hypopharyngeal carcinoma. Cancer. 1972;29(1):117–21.CrossRefGoogle Scholar
  39. 39.
    Zhao M, Sano D, Pickering CR, Jasser SA, Henderson YC, Clayman GL, et al. Assembly and initial characterization of a panel of 85 genomically validated cell lines from diverse head and neck tumor sites. Clin Cancer Res. 2011;17(23):7248–64.CrossRefGoogle Scholar
  40. 40.
    Nichols AC, Yoo J, Palma DA, Fung K, Franklin JH, Koropatnick J, et al. Frequent mutations in TP53 and CDKN2A found by next-generation sequencing of head and neck cancer cell lines. Arch Otolaryngol Head Neck Surg. 2012;138(8):732–9.CrossRefGoogle Scholar
  41. 41.
    Lin CJ, Grandis JR, Carey TE, Gollin SM, Whiteside TL, Koch WM, et al. Head and neck squamous cell carcinoma cell lines: established models and rationale for selection. Head Neck. 2007;29(2):163–88.CrossRefGoogle Scholar
  42. 42.
    Wang J, Pan XL, Ding LJ, Liu DY, Da-Peng L, Jin T. Aberrant expression of Beclin-1 and LC3 correlates with poor prognosis of human hypopharyngeal squamous cell carcinoma. PLoS One. 2013;8(7):e69038.CrossRefGoogle Scholar
  43. 43.
    Tang Z, Li J, Shen Q, Feng J, Liu H, Wang W, et al. Contribution of upregulated dipeptidyl peptidase 9 (DPP9) in promoting tumoregenicity, metastasis and the prediction of poor prognosis in non-small cell lung cancer (NSCLC). Int J Cancer. 2017;140(7):1620–32.CrossRefGoogle Scholar
  44. 44.
    Bresnick AR, Weber DJ, Zimmer DB. S100 proteins in cancer. Nat Rev Cancer. 2015;15(2):96–109.CrossRefGoogle Scholar
  45. 45.
    Ismail TM, Bennett D, Platt-Higgins AM, Al-Medhity M, Barraclough R, Rudland PS. S100A4 Elevation Empowers Expression of Metastasis Effector Molecules in Human Breast Cancer. Cancer Res. 2017;77(3):780–9.CrossRefGoogle Scholar
  46. 46.
    Fabris L, Cadamuro M, Moserle L, Dziura J, Cong X, Sambado L, et al. Nuclear expression of S100A4 calcium-binding protein increases cholangiocarcinoma invasiveness and metastasization. Hepatology. 2011;54(3):890–9.CrossRefGoogle Scholar
  47. 47.
    Kikuchi N, Horiuchi A, Osada R, Imai T, Wang C, Chen X, et al. Nuclear expression of S100A4 is associated with aggressive behavior of epithelial ovarian carcinoma: an important autocrine/paracrine factor in tumor progression. Cancer Sci. 2006;97(10):1061–9.CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Jianing Xu
    • 1
  • Neil Gross
    • 2
  • Yuanwei Zang
    • 3
  • Shengda Cao
    • 1
  • Feilong Yang
    • 3
  • Zheng Yang
    • 4
  • Wenbin Yu
    • 5
    Email author
  • Dapeng Lei
    • 1
  • Xinliang Pan
    • 1
    Email author
  1. 1.NHC Key Laboratory of Otorhinolaryngology, Department of Otorhinolaryngology, Qilu HospitalShandong UniversityJinanChina
  2. 2.Department of Head and Neck SurgeryThe University of Texas MD Anderson Cancer CenterHoustonUSA
  3. 3.Department of Urology,Qilu HospitalShandong UniversityJinanChina
  4. 4.Key Laboratory of Otolaryngology Head and Neck Surgery, Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Beijing Institute of Otolaryngology, Ministry of EducationCapital Medical UniversityBeijingChina
  5. 5.Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Head and Neck SurgeryPeking University Cancer Hospital and InstituteBeijingChina

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