Fascin is a circulating tumor marker for head and neck cancer as determined by a proteomic analysis of interstitial fluid from the tumor microenvironment

Li-Yu Lee; Yin-Ju Chen; Ya-Ching Lu; Chun-Ta Liao; I-How Chen; Joseph Tung-Chieh Chang; Yu-Chen Huang; Wen-Ho Chen; Chi-Che Huang; Chi-Ying Tsai; Ann-Joy Cheng

doi:10.1515/cclm-2014-1016

Article

Fascin is a circulating tumor marker for head and neck cancer as determined by a proteomic analysis of interstitial fluid from the tumor microenvironment

Li-Yu Lee , Yin-Ju Chen , Ya-Ching Lu , Chun-Ta Liao , I-How Chen , Joseph Tung-Chieh Chang , Yu-Chen Huang , Wen-Ho Chen , Chi-Che Huang , Chi-Ying Tsai and Ann-Joy Cheng

Published/Copyright: March 10, 2015

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From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 53 Issue 10

Abstract

Background: Head and neck cancer (HNC) is a prevalent cancer worldwide; however, clinically useful tumor markers for HNC have not been identified. Here, we aimed to identify secretory proteins from the tumor microenvironment as candidate circulating tumor markers.

Methods: Samples derived from seven pairs of tumor interstitial fluid (TIF) and normal interstitial fluid (NIF) samples from patients with HNC were analyzed. The proteomes were determined by gel-based-mass-spectrometry proteomic methods. The most up-regulated protein, fascin was confirmed in the cancer tissues and cell culture supernatant by immunoblotting and immunohistochemistry assays. Serum fascin was determined in 40 HNC and 40 normal individuals by ELISA.

Results: After proteomics analysis, 189 peptides were identified, corresponding to 75 proteins. Of the 21 proteins which were identified more than twice, five up-regulated proteins identified most frequently including fascin. The most elevated fascin was over-expressed in cancer tissues and cell culture supernatant. Serum fascin was significantly up-regulated in the cancer patients (p<0.001) and correlated with pathological lymph node metastasis (p=0.022). To assess the diagnostic efficacy, serum levels of fascin and another potential biomarker SCCA were determined. Fascin showed a high predictable value with an area under the curve (AUC) of 0.808 (95% CI 0.723–0.901) in the receiver operator curve (ROC), compared to 0.501 (95% CI 0.378–0.634) for SCCA.

Conclusions: We have identified 75 potential circulating tumor markers associated with HNC, including fascin. Serum fascin could discriminate cancer patients from healthy individuals; thus, it may serve as a circulating biomarker for HNC.

Keywords: circulating tumor markers; fascin; head and neck cancer; tumor interstitial fluid

Corresponding authors: Ann-Joy Cheng, PhD, Department of Medical Biotechnology, College of Medicine, Chang Gung University, 259 Wen-Hwa 1st Road, Taoyuan 333, Taiwan, Phone: +886 3 2118800 ex 5085, Fax: +886 3 2118247, E-mail: ajchen@mail.cgu.edu.tw; and Joseph Tung-Chieh Chang, MD, Department of Radiation Oncology, Chang Gung Memorial Hospital-Linko, 5 Fu-Shin Road, Toayuan 333, Taiwan, E-mail: jtchang@adm.cgmh.org.tw,

aLi-Yu Lee and Yin-Ju Chen contributed equally.

Acknowledgments

This study was supported by grants from Chang Gung Memorial Hospital (CMRPG360843) and National Science Counsel of Taiwan (NSC-101-2314-B- 182A-121-MY3, NSC-101-2632-B-182-001-MY3).

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Financial support: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

References

1. Chen YJ, Chang JT, Liao CT, Wang HM, Yen TC, Chiu CC, et al. Head and neck cancer in the betel quid chewing area: recent advances in molecular carcinogenesis. Cancer Sci 2008;99:1507–14.10.1111/j.1349-7006.2008.00863.xSearch in Google Scholar PubMed

2. Warnakulasuriya S. Global epidemiology of oral and oropharyngeal cancer. Oral Oncol 2009;45:309–16.10.1016/j.oraloncology.2008.06.002Search in Google Scholar PubMed

3. Saman DM. A review of the epidemiology of oral and pharyngeal carcinoma: update. Head Neck Oncol 2012;4:1.10.1186/1758-3284-4-1Search in Google Scholar PubMed PubMed Central

4. Anderson NL, Anderson NG. The human plasma proteome: history, character, and diagnostic prospects. Mol Cell Proteomics 2002;1;11:845–67.10.1074/mcp.R200007-MCP200Search in Google Scholar PubMed

5. Cheng AJ, Chen LC, Chien KY, Chen YJ, Chang JT, Wang HM, et al. Oral cancer plasma tumor marker identified with bead-based affinity-fractionated proteomic technology. Clin Chem 2005;51:2236–44.10.1373/clinchem.2005.052324Search in Google Scholar PubMed

6. Maurya P, Meleady P, Dowling P, Clynes M. Proteomic approaches for serum biomarker discovery in cancer. Anticancer Res 2007;27:1247–55.Search in Google Scholar

7. Stahl-Zeng J, Lange V, Ossola R, Eckhardt K, Krek W, Aebersold R, et al. High sensitivity detection of plasma proteins by multiple reaction monitoring of N-glycosites. Mol Cell Proteomics 2007;6:1809–17.10.1074/mcp.M700132-MCP200Search in Google Scholar PubMed

8. Chang JT, Chen LC, Wei SY, Chen YJ, Wang HM, Liao CT, et al. Increase diagnostic efficacy by combined use of fingerprint markers in mass spectrometry – plasma peptidomes from nasopharyngeal cancer patients for example. Clin Biochem 2006;39:1144–51.10.1016/j.clinbiochem.2006.08.010Search in Google Scholar PubMed

9. Bruno A, Pagani A, Magnani E, Rossi T, Noonan DM, Cantelmo AR, et al. Inflammatory angiogenesis and the tumor microenvironment as targets for cancer therapy and prevention. Cancer Treat Res 2014;159:401–26.10.1007/978-3-642-38007-5_23Search in Google Scholar PubMed

10. Marcus ME, Leonard JN. FedExosomes: engineering therapeutic biological nanoparticles that truly deliver. Pharmaceuticals (Basel) 2013;6:659–80.10.3390/ph6050659Search in Google Scholar PubMed PubMed Central

11. Shieh AC, Swartz MA. Regulation of tumor invasion by interstitial fluid flow. Phys Biol 2011;1:015012.Search in Google Scholar

12. Shieh AC, Rozansky HA, Hinz B, Swartz MA. Tumor cell invasion is promoted by interstitial flow-induced matrix priming by stromal fibroblasts. Cancer Res 2011;3:790–800.10.1158/0008-5472.CAN-10-1513Search in Google Scholar PubMed

13. Celis JE, Gromov P, Cabezon T, Moreira JM, Ambartsumian N, Sandelin K, et al. Proteomic characterization of the interstitial fluid perfusing the breast tumor microenvironment: a novel resource for biomarker and therapeutic target discovery. Mol Cell Proteomics 2004;3:327–44.10.1074/mcp.M400009-MCP200Search in Google Scholar PubMed

14. Cabezon T, Celis JE, Skibshoj I, Klingelhofer J, Grigorian M, Gromov P, et al. Expression of S100A4 by a variety of cell types present in the tumor microenvironment of human breast cancer. Int J Cancer 2007;121:1433–44.10.1002/ijc.22850Search in Google Scholar PubMed

15. Gromov P, Gromova I, Bunkenborg J, Cabezon T, Moreira JM, Timmermans-Wielenga V, et al. Up-regulated proteins in the fluid bathing the tumour cell microenvironment as potential serological markers for early detection of cancer of the breast. Mol Oncol 2010;4:65–89.10.1016/j.molonc.2009.11.003Search in Google Scholar PubMed PubMed Central

16. Wang TH, Chao A, Tsai CL, Chang CL, Chen SH, Lee YS, et al. Stress-induced phosphoprotein 1 as a secreted biomarker for human ovarian cancer promotes cancer cell proliferation. Mol Cell Proteomics 2010;9:1873–84.10.1074/mcp.M110.000802Search in Google Scholar PubMed PubMed Central

17. Lin TY, Chang JT, Wang HM, Chan SH, Chiu CC, Lin CY, et al. Proteomics of the radioresistant phenotype in head-and-neck cancer: Gp96 as a novel prediction marker and sensitizing target for radiotherapy. Int J Radiat Oncol Biol Phys 2010;78:246–56.10.1016/j.ijrobp.2010.03.002Search in Google Scholar PubMed

18. Chen YJ, Lee LY, Chao YK, Chang JT, Lu YC, Li HF, et al. DSG3 facilitates cancer cell growth and invasion through the DSG3-plakoglobin-TCF/LEF-Myc/cyclin D1/MMP signaling pathway. PLoS One 2013;8:e64088.10.1371/journal.pone.0064088Search in Google Scholar PubMed PubMed Central

19. Lu YC, Chen YJ, Wang HM, Tsai CY, Chen WH, Huang YC, et al. Oncogenic function and early detection potential of miRNA-10b in oral cancer as identified by microRNA profiling. Cancer Prev Res (Phila) 2012;5:665–74.10.1158/1940-6207.CAPR-11-0358Search in Google Scholar PubMed

20. Chang JT, Wong FH, Liao CT, Chen IH, Wang HM, Cheng AJ. Enzyme immunoassay for serum autoantibody to survivin and its findings in head-and-neck cancer patients. Clin Chem 2004;50:1261–4.10.1373/clinchem.2003.026666Search in Google Scholar PubMed

21. Potra F, Liu X, Seillier-Moiseiwitsch F, Roy A, Hang Y, Marten M, et al. Protein image alignment via piecewise affine transformations. J Comput Biol 2006;13:614–30.10.1089/cmb.2006.13.614Search in Google Scholar PubMed

22. Feng XY, Li JH, Li JZ, Han ZX, Xing RD. Serum SCCA, Cyfra 21-1, EGFR and Cyclin D1 levels in patients with oral squamous cell carcinoma. Int J Biol Markers 2010;25:93–8.10.1177/172460081002500206Search in Google Scholar

23. Einarsson R. Squamous cell carcinoma antigen (SCCA) isomers – markers for squamous cell carcinoma. Adv Clin Exp Med 2005;14:643–8.Search in Google Scholar

24. Xiao T, Ying W, Li L, Hu Z, Ma Y, Jiao L, et al. An approach to studying lung cancer-related proteins in human blood. Mol Cell Proteomics 2005;4:1480–6.10.1074/mcp.M500055-MCP200Search in Google Scholar PubMed

25. Hu S, Arellano M, Boontheung P, Wang J, Zhou H, Jiang J, et al. Salivary proteomics for oral cancer biomarker discovery. Clin Cancer Res 2008;14:6246–52.10.1158/1078-0432.CCR-07-5037Search in Google Scholar PubMed PubMed Central

26. Wu CC, Peng PH, Chang YT, Huang YS, Chang KP, Hao SP, et al. Identification of potential serum markers for nasopharyngeal carcinoma from a xenografted mouse model using Cy-dye labeling combined with three-dimensional fractionation. Proteomics 2008;8:3605–20.10.1002/pmic.200701034Search in Google Scholar PubMed

27. Coutinho-Camillo CM, Lourenco SV, Nishimoto IN, Kowalski LP, Soares FA. Nucleophosmin, p53, and Ki-67 expression patterns on an oral squamous cell carcinoma tissue microarray. Hum Pathol 2010;41:1079–86.10.1016/j.humpath.2009.12.010Search in Google Scholar PubMed

28. Tseng MY, Liu SY, Chen HR, Wu YJ, Chiu CC, Chan PT, et al. Serine protease inhibitor (SERPIN) B1 promotes oral cancer cell motility and is over-expressed in invasive oral squamous cell carcinoma. Oral Oncol 2009;45:771–6.10.1016/j.oraloncology.2008.11.013Search in Google Scholar PubMed

29. Lin CY, Lin TY, Wang HM, Huang SF, Fan KH, Liao CT, et al. GP96 is over-expressed in oral cavity cancer and is a poor prognostic indicator for patients receiving radiotherapy. Radiat Oncol 2011;6:136.10.1186/1748-717X-6-136Search in Google Scholar PubMed PubMed Central

30. Arjonen A, Kaukonen R, Ivaska J. Filopodia and adhesion in cancer cell motility. Cell Adh Migr 2011;5:421–30.10.4161/cam.5.5.17723Search in Google Scholar PubMed PubMed Central

31. Machesky LM, Li A. Fascin: invasive filopodia promoting metastasis. Commun Integr Biol 2010;3:263–70.10.4161/cib.3.3.11556Search in Google Scholar PubMed PubMed Central

32. Durmaz A, Kurt B, Ongoru O, Karahatay S, Gerek M, Yalcin S. Significance of fascin expression in laryngeal squamous cell carcinoma. J Laryngol Otol 2010;124:194–8.10.1017/S0022215109991630Search in Google Scholar PubMed

33. Zou J, Yang H, Chen F, Zhao H, Lin P, Zhang J, et al. Prognostic significance of fascin-1 and E-cadherin expression in laryngeal squamous cell carcinoma. Eur J Cancer Prev 2010;19:11–7.10.1097/CEJ.0b013e32832f9aa6Search in Google Scholar PubMed

34. Gao W, Zhang C, Feng Y, Chen G, Wen S, Huangfu H, et al. Fascin-1, ezrin and paxillin contribute to the malignant progression and are predictors of clinical prognosis in laryngeal squamous cell carcinoma. PLoS One 2012;7:e50710.10.1371/journal.pone.0050710Search in Google Scholar PubMed PubMed Central

35. Alam H, Bhate AV, Gangadaran P, Sawant SS, Salot S, Sehgal L, et al. Fascin overexpression promotes neoplastic progression in oral squamous cell carcinoma. BMC Cancer 2012;12:32.10.1186/1471-2407-12-32Search in Google Scholar PubMed PubMed Central

36. Rink M, Chun FK, Robinson B, Sun M, Karakiewicz PI, Bensalah K, et al. Tissue-based molecular markers for renal cell carcinoma. Minerva Urol Nefrol 2011;63:293–308.Search in Google Scholar

37. Kulasingam V, Diamandis EP. Fascin-1 is a novel biomarker of aggressiveness in some carcinomas. BMC Med 2013;11:53.10.1186/1741-7015-11-53Search in Google Scholar PubMed PubMed Central

38. Teng Y, Xu S, Yue W, Ma L, Zhang L, Zhao X, et al. Serological investigation of the clinical significance of fascin in non-small-cell lung cancer. Lung Cancer 2013;82:346.10.1016/j.lungcan.2013.08.017Search in Google Scholar PubMed

Received: 2014-10-17

Accepted: 2015-1-20

Published Online: 2015-3-10

Published in Print: 2015-9-1

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Articles in the same Issue

https://doi.org/10.1515/cclm-2014-1016

Keywords for this article

circulating tumor markers; fascin; head and neck cancer; tumor interstitial fluid