Expression of TMPRSS4 in non-small cell lung cancer and its modulation by hypoxia

Nguyen,Tri-Hung; Weber,William; Havari,Evis; Connors,Timothy; Bagley,Rebecca  G.; McLaren,Rajashree; Nambiar,Prashant  R.; Madden,Stephen   L.; Teicher,Beverly  A.; Roberts,Bruce; Kaplan,Johanne; Shankara,Srinivas

doi:10.3892/ijo.2012.1513

Expression of TMPRSS4 in non-small cell lung cancer and its modulation by hypoxia

Authors:
- Tri-Hung Nguyen
- William Weber
- Evis Havari
- Timothy Connors
- Rebecca G. Bagley
- Rajashree McLaren
- Prashant R. Nambiar
- Stephen L. Madden
- Beverly A. Teicher
- Bruce Roberts
- Johanne Kaplan
- Srinivas Shankara
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Affiliations: Genzyme Corporation, Framingham, MA 01701, USA
Published online on: June 12, 2012 https://doi.org/10.3892/ijo.2012.1513
Pages: 829-838
Copyright: © Nguyen et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Overexpression of TMPRSS4, a cell surface-associated transmembrane serine protease, has been reported in pancreatic, colorectal and thyroid cancers, and has been implicated in tumor cell migration and metastasis. Few reports have investigated both TMPRSS4 gene expression levels and the protein products. In this study, quantitative RT-PCR and protein staining were used to assess TMPRSS4 expression in primary non-small cell lung carcinoma (NSCLC) tissues and in lung tumor cell lines. At the transcriptional level, TMPRSS4 message was significantly elevated in the majority of human squamous cell and adenocarcinomas compared with normal lung tissues. Staining of over 100 NSCLC primary tumor and normal specimens with rabbit polyclonal anti-TMPRSS4 antibodies confirmed expression at the protein level in both squamous cell and adenocarcinomas with little or no staining in normal lung tissues. Human lung tumor cell lines expressed varying levels of TMPRSS4 mRNA in vitro. Interestingly, tumor cell lines with high levels of TMPRSS4 mRNA failed to show detectable TMPRSS4 protein by either immunoblotting or flow cytometry. However, protein levels were increased under hypoxic culture conditions suggesting that hypoxia within the tumor microenvironment may upregulate TMPRSS4 protein expression in vivo. This was supported by the observation of TMPRSS4 protein in xenograft tumors derived from the cell lines. In addition, staining of human squamous cell carcinoma samples for carbonic anhydrase IX (CAIX), a hypoxia marker, showed TMPRSS4 positive cells adjacent to CAIX positive cells. Overall, these results indicate that the cancer-associated TMPRSS4 protein is overexpressed in NSCLC and may represent a potential therapeutic target.

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1	Group UCSW: United States Cancer Statistics: 1999–2005 Incidence and mortality Web-based report, 2009.
2	Sher YP, Shih JY, Yang PC, et al: Prognosis of non-small cell lung cancer patients by detecting circulating cancer cells in the peripheral blood with multiple marker genes. Clin Cancer Res. 11:173–179. 2005.PubMed/NCBI
3	Petersen I and Petersen S: Towards a genetic-based classification of human lung cancer. Anal Cell Pathol. 22:111–121. 2001. View Article : Google Scholar : PubMed/NCBI
4	Lu Y, Lemon W, Liu PY, et al: A gene expression signature predicts survival of patients with stage I non-small cell lung cancer. PLoS Med. 3:e4672006. View Article : Google Scholar : PubMed/NCBI
5	Boeri M, Verri C, Conte D, et al: MicroRNA signatures in tissues and plasma predict development and prognosis of computed tomography detected lung cancer. Proc Natl Acad Sci USA. 108:3713–3718. 2011. View Article : Google Scholar : PubMed/NCBI
6	Netzel-Arnett S, Hooper JD, Szabo R, et al: Membrane anchored serine proteases: a rapidly expanding group of cell surface proteolytic enzymes with potential roles in cancer. Cancer Metastasis Rev. 22:237–258. 2003. View Article : Google Scholar : PubMed/NCBI
7	Gress TM, Wallrapp C, Frohme M, et al: Identification of genes with specific expression in pancreatic cancer by cDNA representational difference analysis. Genes Chromosomes Cancer. 19:97–103. 1997. View Article : Google Scholar : PubMed/NCBI
8	Wallrapp C, Hahnel S, Muller-Pillasch F, et al: A novel transmembrane serine protease (TMPRSS3) overexpressed in pancreatic cancer. Cancer Res. 60:2602–2606. 2000.PubMed/NCBI
9	Butterworth MB, Edinger RS, Frizzell RA and Johnson JP: Regulation of the epithelial sodium channel by membrane trafficking. Am J Physiol Renal Physiol. 296:F10–F24. 2009. View Article : Google Scholar : PubMed/NCBI
10	Bhalla V and Hallows KR: Mechanisms of ENaC regulation and clinical implications. J Am Soc Nephrol. 19:1845–1854. 2008. View Article : Google Scholar : PubMed/NCBI
11	Grutzmann R, Pilarsky C, Ammerpohl O, et al: Gene expression profiling of microdissected pancreatic ductal carcinomas using high-density DNA microarrays. Neoplasia. 6:611–622. 2004. View Article : Google Scholar : PubMed/NCBI
12	Iacobuzio-Donahue CA, Ashfaq R, Maitra A, et al: Highly expressed genes in pancreatic ductal adenocarcinomas: a comprehensive characterization and comparison of the transcription profiles obtained from three major technologies. Cancer Res. 63:8614–8622. 2003.
13	Kebebew E, Peng M, Reiff E, Duh QY, Clark OH and McMillan A: ECM1 and TMPRSS4 are diagnostic markers of malignant thyroid neoplasms and improve the accuracy of fine needle aspiration biopsy. Ann Surg. 242:353–361. 2005.PubMed/NCBI
14	Ma XJ, Patel R, Wang X, et al: Molecular classification of human cancers using a 92-gene real-time quantitative polymerase chain reaction assay. Arch Pathol Lab Med. 130:465–473. 2006.PubMed/NCBI
15	Riker AI, Enkemann SA, Fodstad O, et al: The gene expression profiles of primary and metastatic melanoma yields a transition point of tumor progression and metastasis. BMC Med Genomics. 1:132008. View Article : Google Scholar : PubMed/NCBI
16	Larzabal L, Nguewa PA, Pio R, et al: Overexpression of TMPRSS4 in non-small cell lung cancer is associated with poor prognosis in patients with squamous histology. Br J Cancer. 105:1608–1614. 2011. View Article : Google Scholar : PubMed/NCBI
17	Jung H, Lee KP, Park SJ, et al: TMPRSS4 promotes invasion, migration and metastasis of human tumor cells by facilitating an epithelial-mesenchymal transition. Oncogene. 27:2635–2647. 2008. View Article : Google Scholar
18	Kim S, Kang HY, Nam EH, et al: TMPRSS4 induces invasion and epithelial-mesenchymal transition through upregulation of integrin {alpha}5 and its signaling pathways. Carcinogenesis. 31:597–606. 2010.PubMed/NCBI
19	Li T, Zeng ZC, Wang L, et al: Radiation enhances long-term metastasis potential of residual hepatocellular carcinoma in nude mice through TMPRSS4-induced epithelial-mesenchymal transition. Cancer Gene Ther. 18:617–626. 2011. View Article : Google Scholar
20	Hermanson GT: Bioconjugate Techniques. Academic Press; San Diego: 2008
21	Kyte J and Doolittle RF: A simple method for displaying the hydropathic character of a protein. J Mol Biol. 157:105–132. 1982. View Article : Google Scholar : PubMed/NCBI
22	Kivela AJ, Parkkila S, Saarnio J, et al: Expression of transmembrane carbonic anhydrase isoenzymes IX and XII in normal human pancreas and pancreatic tumours. Histochem Cell Biol. 114:197–204. 2000.PubMed/NCBI
23	Juhasz M, Chen J, Lendeckel U, et al: Expression of carbonic anhydrase IX in human pancreatic cancer. Aliment Pharmacol Ther. 18:837–846. 2003. View Article : Google Scholar : PubMed/NCBI
24	Nguyen TH, Havari E, Connors T, et al: Cancer cells expressing TMPRSS4 colocalized with carbonic anhydrase IX (CAIX)-positive cells in lung and pancreatic carcinomas. Mol Cancer Ther. 8:C1662009. View Article : Google Scholar
25	Jia JB, Wang WQ, Sun HC, et al: A novel tripeptide, tyroserleutide, inhibits irradiation-induced invasiveness and metastasis of hepatocellular carcinoma in nude mice. Invest New Drugs. 29:861–872. 2010. View Article : Google Scholar : PubMed/NCBI
26	Giatromanolaki A, Koukourakis MI, Sivridis E, et al: Expression of hypoxia-inducible carbonic anhydrase-9 relates to angiogenic pathways and independently to poor outcome in non-small cell lung cancer. Cancer Res. 61:7992–7998. 2001.PubMed/NCBI
27	Egeblad M and Werb Z: New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2:161–174. 2002. View Article : Google Scholar : PubMed/NCBI
28	Szabo R, Wu Q, Dickson RB, Netzel-Arnett S, Antalis TM and Bugge TH: Type II transmembrane serine proteases. Thromb Haemost. 90:185–193. 2003.PubMed/NCBI
29	Chaipan C, Kobasa D, Bertram S, et al: Proteolytic activation of the 1918 influenza virus hemagglutinin. J Virol. 83:3200–3211. 2009. View Article : Google Scholar : PubMed/NCBI
30	Garcia-Caballero A, Dang Y, He H and Stutts MJ: ENaC proteolytic regulation by channel-activating protease 2. J Gen Physiol. 132:521–535. 2008. View Article : Google Scholar : PubMed/NCBI
31	Kellenberger S and Schild L: Epithelial sodium channel/degenerin family of ion channels: a variety of functions for a shared structure. Physiol Rev. 82:735–767. 2002.PubMed/NCBI
32	Vuagniaux G, Vallet V, Jaeger NF, Hummler E and Rossier BC: Synergistic activation of ENaC by three membrane-bound channel-activating serine proteases (mCAP1, mCAP2, and mCAP3) and serum- and glucocorticoid-regulated kinase (Sgk1) in Xenopus Oocytes. J Gen Physiol. 120:191–201. 2002. View Article : Google Scholar
33	Passero CJ, Mueller GM, Myerburg MM, Carattino MD, Hughey RP and Kleyman TR: TMPRSS4-dependent activation of the epithelial sodium channel requires cleavage of the gamma subunit distal to the furin cleavage site. Am J Physiol Renal Physiol. 302:F1–F8. 2012. View Article : Google Scholar : PubMed/NCBI
34	Rossier BC: The epithelial sodium channel: activation by membrane-bound serine proteases. Proc Am Thorac Soc. 1:4–9. 2004. View Article : Google Scholar : PubMed/NCBI
35	Planes C and Caughey GH: Regulation of the epithelial Na⁺ channel by peptidases. Curr Top Dev Biol. 78:23–46. 2007. View Article : Google Scholar
36	Matsushita K, McCray PB Jr, Sigmund RD, Welsh MJ and Stokes JB: Localization of epithelial sodium channel subunit mRNAs in adult rat lung by in situ hybridization. Am J Physiol. 271:L332–L339. 1996.PubMed/NCBI
37	Yamamura H, Ugawa S, Ueda T, Nagao M and Shimada S: Protons activate the delta-subunit of the epithelial Na⁺ channel in humans. J Biol Chem. 279:12529–12534. 2004. View Article : Google Scholar : PubMed/NCBI
38	Yamamura H, Ugawa S, Ueda T and Shimada S: Expression analysis of the epithelial Na⁺ channel delta subunit in human melanoma G-361 cells. Biochem Biophys Res Commun. 366:489–492. 2008.PubMed/NCBI
39	Ji HL and Benos DJ: Degenerin sites mediate proton activation of deltabetagamma-epithelial sodium channel. J Biol Chem. 279:26939–26947. 2004. View Article : Google Scholar : PubMed/NCBI
40	Wodopia R, Ko HS, Billian J, Wiesner R, Bartsch P and Mairbaurl H: Hypoxia decreases proteins involved in epithelial electrolyte transport in A549 cells and rat lung. Am J Physiol Lung Cell Mol Physiol. 279:L1110–L1119. 2000.PubMed/NCBI
41	Bouvry D, Planes C, Malbert-Colas L, Escabasse V and Clerici C: Hypoxia-induced cytoskeleton disruption in alveolar epithelial cells. Am J Respir Cell Mol Biol. 35:519–527. 2006. View Article : Google Scholar : PubMed/NCBI
42	Kebebew E, Peng M, Reiff E and McMillan A: Diagnostic and extent of disease multigene assay for malignant thyroid neoplasms. Cancer. 106:2592–2597. 2006. View Article : Google Scholar : PubMed/NCBI