Overexpression of fibronectin confers cell adhesion‑mediated drug resistance (CAM-DR) against 5-FU in oral squamous cell carcinoma cells

Nakagawa,Yoshihiro; Nakayama,Hideki; Nagata,Masashi; Yoshida,Ryoji; Kawahara,Kenta; Hirosue,Akiyuki; Tanaka,Takuya; Yuno,Akira; Matsuoka,Yuichiro; Kojima,Taku; Yoshitake,Yoshihiro; Hiraki,Akimitsu; Shinohara,Masanori

doi:10.3892/ijo.2014.2265

Overexpression of fibronectin confers cell adhesion‑mediated drug resistance (CAM-DR) against 5-FU in oral squamous cell carcinoma cells

Authors:
- Yoshihiro Nakagawa
- Hideki Nakayama
- Masashi Nagata
- Ryoji Yoshida
- Kenta Kawahara
- Akiyuki Hirosue
- Takuya Tanaka
- Akira Yuno
- Yuichiro Matsuoka
- Taku Kojima
- Yoshihiro Yoshitake
- Akimitsu Hiraki
- Masanori Shinohara
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Affiliations: Department of Oral and Maxillofacial Surgery, Faculty of Life Sciences, Kumamoto University, Chuo-ku, Kumamoto 860-8556, Japan
Published online on: January 21, 2014 https://doi.org/10.3892/ijo.2014.2265
Pages: 1376-1384

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Abstract

The tumor-associated microenvironment has been shown to protect tumor cells from treatment, and the extracellular matrix (ECM) is known to affect drug resistance as a key regulator of the tumor microenvironment. However, little is known about cell adhesion-mediated drug resistance (CAM‑DR) due to cell-ECM contact in patients with oral squamous cell carcinoma (OSCC). In the present study, we evaluated the ECM molecule fibronectin (FN) using DNA microarray data obtained from parental and 5-FU-resistant OSCC cell lines. We investigated the effects of cell adhesion to FN on 5-FU resistance in OSCC cells and examined the activation of FN receptor β1 integrin‑mediated survival regulators such as ILK, Akt and NF-κB. In addition, we investigated whether FNIII14, a 22-mer peptide derived from FN that potently prevents β1 integrin-mediated adhesion to FN, could overcome CAM-DR against 5-FU in OSCC cells and examined the activation of survival regulators and apoptosis-related molecules. Consequently, we obtained the following results. FN was extracellularly overexpressed in the 5-FU-resistant cells compared with that observed in the 5-FU-sensitive cells. Cell adhesion to FN enhanced 5-FU resistance and activated integrin-mediated ILK/Akt/NF-κB survival signaling in the 5-FU-resistant OSCC cells. Furthermore, the inhibition of cell adhesion to FN by FNIII14 enhanced chemosensitivity to 5-FU and apoptosis by suppressing ILK/Akt/NF-κB signaling in the 5-FU-resistant cells. These novel findings demonstrate that FN is a potentially useful biomarker and therapeutic target for improving the treatment of OSCC, particularly in the setting of 5-FU resistance.

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1.	Lawrence TS, Tepper JE and Blackstock AW: Fluoropyrimidine-radiation interactions in cells and tumors. Semin Radiat Oncol. 7:260–266. 1997. View Article : Google Scholar : PubMed/NCBI
2.	Pignon JP, Bourhis J, Domenge C and Designe L: Chemotherapy added to locoregional treatment for head and neck squamous-cell carcinoma: three meta-analyses of updated individual data. MACH-NC Collaborative Group. Meta-analysis of chemo-therapy on head and neck cancer. Lancet. 355:949–955. 2000. View Article : Google Scholar
3.	Adelstein DJ, Saxton JP, Rybicki LA, et al: Multiagent concurrent chemoradiotherapy for locoregionally advanced squamous cell head and neck cancer: mature results from a single institution. J Clin Oncol. 24:1064–1071. 2006. View Article : Google Scholar : PubMed/NCBI
4.	Tsukuda M, Ishitoya J, Matsuda H, et al: Randomized controlled phase II comparison study of concurrent chemoradiotherapy with docetaxel, cisplatin, and 5-fluorouracil versus CCRT with cisplatin, 5-fluorouracil, methotrexate and leucovorin in patients with locally advanced squamous cell carcinoma of the head and neck. Cancer Chemother Pharmacol. 66:729–736. 2010.
5.	Shingaki S, Takada M, Sasai K, et al: Impact of lymph node metastasis on the pattern of failure and survival in oral carcinomas. Am J Surg. 185:278–284. 2003. View Article : Google Scholar : PubMed/NCBI
6.	Bell RB, Kademani D, Homer L, Dierks EJ and Potter BE: Tongue cancer: Is there a difference in survival compared with other subsites in the oral cavity? J Oral Maxillofac Surg. 65:229–236. 2007. View Article : Google Scholar : PubMed/NCBI
7.	Colevas AD: Chemotherapy options for patients with metastatic or recurrent squamous cell carcinoma of the head and neck. J Clin Oncol. 24:2644–2652. 2006. View Article : Google Scholar
8.	Gibson MK, Li Y, Murphy B, et al: Randomized phase III evaluation of cisplatin plus fluorouracil versus cisplatin plus paclitaxel in advanced head and neck cancer (E1395): an intergroup trial of the Eastern Cooperative Oncology Group. J Clin Oncol. 23:3562–3567. 2005. View Article : Google Scholar : PubMed/NCBI
9.	Kang HC, Kim IJ, Park JH, et al: Identification of genes with differential expression in acquired drug-resistant gastric cancer cells using high-density oligonucleotide microarrays. Clin Cancer Res. 10:272–284. 2004. View Article : Google Scholar
10.	Herrmann R: 5-Fluorouracil in colorectal cancer, a never ending story. Ann Oncol. 7:551–552. 1996. View Article : Google Scholar : PubMed/NCBI
11.	Yoo BC, Jeon E, Hong SH, Shin YK, Chang HJ and Park JG: Metabotropic glutamate receptor 4-mediated 5-fluorouracil resistance in a human colon cancer cell line. Clin Cancer Res. 10:4176–4184. 2004. View Article : Google Scholar : PubMed/NCBI
12.	Wang W, Cassidy J, O’Brien V, Ryan KM and Collie-Duguid E: Mechanistic and predictive profiling of 5-fluorouracil resistance in human cancer cells. Cancer Res. 64:8167–8176. 2004. View Article : Google Scholar : PubMed/NCBI
13.	Petersen SL, Peyton M, Minna JD and Wang X: Overcoming cancer cell resistance to Smac mimetic induced apoptosis by modulating cIAP-2 expression. Proc Natl Acad Sci USA. 107:11936–11941. 2010. View Article : Google Scholar : PubMed/NCBI
14.	Meads MB, Gatenby RA and Dalton WS: Environment-mediated drug resistance: a major contributor to minimal residual disease. Nat Rev Cancer. 9:665–674. 2009. View Article : Google Scholar : PubMed/NCBI
15.	Denys H, Braems G, Lambein K, et al: The extracellular matrix regulates cancer progression and therapy response: implications for prognosis and treatment. Curr Pharm Des. 15:1373–1384. 2009. View Article : Google Scholar : PubMed/NCBI
16.	Cukierman E and Bassi DE: The mesenchymal tumor microenvironment: a drug-resistant niche. Cell Adh Migr. 6:285–296. 2012. View Article : Google Scholar : PubMed/NCBI
17.	Hynes RO: Integrins: bidirectional, allosteric signaling machines. Cell. 110:673–687. 2002. View Article : Google Scholar : PubMed/NCBI
18.	Aoudjit F and Vuori K: Integrin signaling inhibits paclitaxel-induced apoptosis in breast cancer cells. Oncogene. 20:4995–5004. 2001. View Article : Google Scholar : PubMed/NCBI
19.	Damiano JS: Integrins as novel drug targets for overcoming innate drug resistance. Curr Cancer Drug Targets. 2:37–43. 2002. View Article : Google Scholar : PubMed/NCBI
20.	Lewis JM, Truong TN and Schwartz MA: Integrins regulate the apoptotic response to DNA damage through modulation of p53. Proc Natl Acad Sci USA. 99:3627–3632. 2002. View Article : Google Scholar : PubMed/NCBI
21.	Sethi T, Rintoul RC, Moore SM, et al: Extracellular matrix proteins protect small cell lung cancer cells against apoptosis: a mechanism for small cell lung cancer growth and drug resistance in vivo. Nat Med. 5:662–668. 1999. View Article : Google Scholar : PubMed/NCBI
22.	Hannigan G, Troussard AA and Dedhar S: Integrin-linked kinase: a cancer therapeutic target unique among its ILK. Nat Rev Cancer. 5:51–63. 2005. View Article : Google Scholar : PubMed/NCBI
23.	Shain KH and Dalton WS: Cell adhesion is a key determinant in de novo multidrug resistance (MDR): new targets for the prevention of acquired MDR. Mol Cancer Ther. 1:69–78. 2001.PubMed/NCBI
24.	Kato R, Ishikawa T, Kamiya S, et al: A new type of anti-metastatic peptide derived from fibronectin. Clin Cancer Res. 8:2455–2462. 2002.PubMed/NCBI
25.	Matsunaga T, Fukai F, Miura S, et al: Combination therapy of an anticancer drug with the FNIII14 peptide of fibronectin effectively overcomes cell adhesion-mediated drug resistance of acute myelogenous leukemia. Leukemia. 22:353–360. 2008. View Article : Google Scholar
26.	Fukai F, Kamiya S, Ohwaki T, et al: The fibronectin-derived anti-adhesive peptide III14-2 suppresses adhesion and apoptosis of leukemic cell lines through down-regulation of protein-tyrosine phosphorylation. Cell Mol Biol (Noisy-le-grand). 46:145–152. 2000.PubMed/NCBI
27.	Nagata M, Nakayama H, Tanaka T, et al: Overexpression of cIAP2 contributes to 5-FU resistance and a poor prognosis in oral squamous cell carcinoma. Br J Cancer. 105:1322–1330. 2011. View Article : Google Scholar : PubMed/NCBI
28.	Bolstad BM, Irizarry RA, Astrand M and Speed TP: A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics. 19:185–193. 2003. View Article : Google Scholar : PubMed/NCBI
29.	Gentleman RC, Carey VJ, Bates DM, et al: Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 5:R802004. View Article : Google Scholar : PubMed/NCBI
30.	Quackenbush J: Microarray data normalization and transformation. Nat Genet. 32(Suppl): 496–501. 2002. View Article : Google Scholar
31.	Fukai F, Hasebe S, Ueki M, et al: Identification of the anti-adhesive site buried within the heparin-binding domain of fibronectin. J Biochem. 121:189–192. 1997.PubMed/NCBI
32.	Hazlehurst LA, Damiano JS, Buyuksal I, Pledger WJ and Dalton WS: Adhesion to fibronectin via beta1 integrins regulates p27kip1 levels and contributes to cell adhesion mediated drug resistance (CAM-DR). Oncogene. 19:4319–4327. 2000. View Article : Google Scholar : PubMed/NCBI
33.	Kouniavsky G, Khaikin M, Zvibel I, et al: Stromal extracellular matrix reduces chemotherapy-induced apoptosis in colon cancer cell lines. Clin Exp Metastasis. 19:55–60. 2002. View Article : Google Scholar : PubMed/NCBI
34.	Uhm JH, Dooley NP, Kyritsis AP, Rao JS and Gladson CL: Vitronectin, a glioma-derived extracellular matrix protein, protects tumor cells from apoptotic death. Clin Cancer Res. 5:1587–1594. 1999.
35.	Matsunaga T, Takemoto N, Sato T, et al: Interaction between leukemic-cell VLA-4 and stromal fibronectin is a decisive factor for minimal residual disease of acute myelogenous leukemia. Nat Med. 9:1158–1165. 2003. View Article : Google Scholar
36.	Helleman J, Jansen MP, Ruigrok-Ritstier K, et al: Association of an extracellular matrix gene cluster with breast cancer prognosis and endocrine therapy response. Clin Cancer Res. 14:5555–5564. 2008. View Article : Google Scholar
37.	Jansen MP, Foekens JA, van Staveren IL, et al: Molecular classification of tamoxifen-resistant breast carcinomas by gene expression profiling. J Clin Oncol. 23:732–740. 2005. View Article : Google Scholar : PubMed/NCBI
38.	Morin PJ: Drug resistance and the microenvironment: nature and nurture. Drug Resist Updat. 6:169–172. 2003. View Article : Google Scholar : PubMed/NCBI
39.	Miyamoto H, Murakami T, Tsuchida K, Sugino H, Miyake H and Tashiro S: Tumor-stroma interaction of human pancreatic cancer: acquired resistance to anticancer drugs and proliferation regulation is dependent on extracellular matrix proteins. Pancreas. 28:38–44. 2004. View Article : Google Scholar
40.	Shain KH, Landowski TH and Dalton WS: The tumor micro-environment as a determinant of cancer cell survival: a possible mechanism for de novo drug resistance. Curr Opin Oncol. 12:557–563. 2000. View Article : Google Scholar : PubMed/NCBI
41.	Persad S, Attwell S, Gray V, et al: Regulation of protein kinase B/Akt-serine 473 phosphorylation by integrin-linked kinase: critical roles for kinase activity and amino acids arginine 211 and serine 343. J Biol Chem. 276:27462–27469. 2001. View Article : Google Scholar
42.	Nicholson KM and Anderson NG: The protein kinase B/Akt signalling pathway in human malignancy. Cell Signal. 14:381–395. 2002. View Article : Google Scholar : PubMed/NCBI
43.	Tan C, Mui A and Dedhar S: Integrin-linked kinase regulates inducible nitric oxide synthase and cyclooxygenase-2 expression in an NF-kappa B-dependent manner. J Biol Chem. 277:3109–3116. 2002. View Article : Google Scholar : PubMed/NCBI
44.	Younes MN, Yigitbasi OG, Yazici YD, et al: Effects of the integrin-linked kinase inhibitor QLT0267 on squamous cell carcinoma of the head and neck. Arch Otolaryngol Head Neck Surg. 133:15–23. 2007. View Article : Google Scholar : PubMed/NCBI