Epidermal growth factor signals regulate dihydropyrimidine dehydrogenase expression in EGFR-mutated non-small-cell lung cancer

English version České info

Background:
It has been shown that epidermal growth factor receptor (EGFR) mutation status is associated with 5-fluorouracil (5-FU) sensitivity in non-small-cell lung cancer (NSCLC). However, the relationship between EGFR mutation status and dihydropyrimidine dehydrogenase (DPD), a 5-FU degrading enzyme, is unknown.

Methods:
We elucidated the crosstalk among the EGFR signal cascade, the DPD gene (DPYD), and DPD protein expression via the transcription factor Sp1 and the effect of EGFR mutation status on the crosstalk.

Results:
In the PC9 (exon19 E746-A750) study, EGF treatment induced up-regulation of both Sp1 and DPD; gefitinib, an EGFR-tyrosine kinase inhibitor (EGFR-TKI), and mithramycin A, a specific Sp-1 inhibitor, suppressed them. Among EGFR-mutated (PC9, HCC827; exon19 E746-A750 and H1975; exon21 L858R, T790M, gefitinib resistant) and -non-mutated (H1437, H1299) cell lines, EGF administration increased DPYD mRNA expression only in mutated cells (p < 0.05). Accordingly, gefitinib inhibited DPD protein expression only in PC9 and HCC827 cells, and mithramycin A inhibited it in EGFR-mutated cell lines, but not in wild-type. FU treatment decreased the level of cell viability more in gefitinib-treated EGFR-TKI sensitive cell lines. Further, combination treatment of FU and mithramycin A suppressed cell viability even in a gefitinib resistant cell line.

Conclusions:
The EGFR signal cascade regulates DPD expression via Sp1 in EGFR mutant cells. These results might be a step towards new therapies targeting Sp1 and DPD in NSCLC with different EGFR mutant status.

Keywords:
Non-small-cell lung cancer Sp1 Dihydropyrimidine dehydrogenase Epidermal growth factor receptor mutation 5-fluorouracil

Autoři: Tetsuro Tominaga ¹; Tomoshi Tsuchiya ¹; Koji Mochinaga ¹; Junichi Arai ¹; Naoya Yamasaki ¹; Keitaro Matsumoto ¹; Takuro Miyazaki ¹; Toshiya Nagasaki ¹; Atsushi Nanashima ²; Kazuhiro Tsukamoto ³; Takeshi Nagayasu ^1*
Působiště autorů: Division of Surgical Oncology, Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki, Nagasaki 85 -8501, Japan. ¹; Department of Surgery, Miyazaki University School of Medicine, 5200 Kihara, Miyazaki, Miyazaki 889-1692, Japan. ²; Department of Pharmacotherapeutics, Nagasaki University Graduate School of Biomedical Science, 1-14 Bunkyo, Nagasaki, Nagasaki 852-8521, Japan. ³
Vyšlo v časopise: BMC Cancer 2016, 16:354
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1186/s12885-016-2392-0

© The Authors. 2016

Open access
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
The electronic version of this article is the complete one and can be found online at: http://bmccancer.biomedcentral.com/articles/10.1186/s12885-016-2392-0

Souhrn

Keywords:
Non-small-cell lung cancer Sp1 Dihydropyrimidine dehydrogenase Epidermal growth factor receptor mutation 5-fluorouracil

Zdroje

1. Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, et al. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med. 2004;350:2129–39.

2. Paez JG, Jänne PA, Lee JC, Tracy S, Greulich H, Gabriel S, et al. EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy. Science. 2004;304:1497–500.

3. Thatcher N, Chang A, Parikh P, Rodriques PJ, Ciuleanu T, Pawel J, et al. Gefitinib plus best supportive care in previously treated patients with refractory advanced non-small-cell lung cancer: results from a randomised, placebo-controlled, multicentre study (Iressa Survival Evaluation in Lung Cancer). Lancet. 2005;366:1527–37.

4. Mitsudomi T, Kosaka T, Endoh H, Horio Y, Hida T, Mori S, et al. Mutations of the epidermal growth factor receptor gene predict prolonged survival after gefitinib treatment in patients with non-small-cell lung cancer with postoperative recurrence. J Clin Oncol. 2005;23:2513–20.

5. Morita S, Hirashima T, Hagiwara K, Hida T, Sunaga K, Sugio A, et al. I-CAMP Study Group Gefitinib combined survival analysis of the mutation positives from the prospective phase II trials ASCO Annual Meeting Proceedings. J Clin Oncol. 2008;26:8101.

6. Mitsudomi T, Morita S, Yatabe Y, Negoro S, Okamoto I, Tsurutani J, et al. Gefitinib versus cisplatin plus docetaxel in patients with non-small-cell lung cancer harbouring mutations of the epidermal growth factor receptor (WJTOG3405): an open label, randomised phase 3 trial. Lancet Oncol. 2010;11:121–8.

7. Maemondo M, Inoue A, Kobayashi K, Sugawara S, Oizumi S, Isobe H, et al. Gefitinib or chemotherapy for non-small-cell lung cancer with mutated EGFR. N Engl J Med. 2010;362:2380–8.

8. Zhou C, Wu YL, Chen G, Feng J, Liu XQ, Wang C, et al. Erlotinib versus chemotherapy as first-line treatment for patients with advanced EGFR mutation-positive non-small-cell lung cancer (OPTIMAL, CTONG-0802): a multicentre, open label, randomised, phase 3 study. Lancet Oncol. 2011;12:735–42.

9. Rosell R, Carcereny E, Gervais R, Vergnenegre A, Massuti B, Felip E, et al. Erlotinib versus standard chemotherapy as first-line treatment for European patients with advanced EGFR mutation-positive non-small-cell lung cancer (EURTAC):a multicenter, open-label, randomized phase 3 trial. Lancet Oncol. 2012;13:239–346.

10. Wu YL, Zhou C, Hu CP, Feng J, Lu S, Huang Y, et al. Afatinib versus cisplatin plus gemcitabine for first-line treatment of Asian patients with advanced non-small-cell lung cancer harbouring EGFR mutations (LUX-Lung6):an open-label, randomised phase 3 trial. Lancet Oncol. 2014;15:213–22.

11. Pez-Ares L, Soulieres D, Melezinek I, Moecks J, Keil L, Mok T, et al. Clinical outcomes in non-small-cell lung cancer patients with EGFR mutations: pooled analysis. J Cell Mol Med. 2010;14:51–69.

12. Jackman D, Pao W, Riely GJ, Engelman JA, Kris MG, Janne PA, et al. Clinical definition of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small-cell lung cancer. J Clin Oncol. 2010;28:357–60.

13. Sequist LV, Yang JC, Yamamoto N, O’Byrne K, Hirsh V, Mok T, et al. Phase III study of afanitib or cisplatin plus pemetrexed in patients with metastatic lung adenocarcinoma with EGFR mutations. J Clin Oncol. 2014;15:213–22.

14. Maring JG, Groen HJ, Wachters FM, Uqes DR, de Vries EG. Genetic factors influencing pyrimidine-antagonist chemotherapy. Pharmacogenomics J. 2005;5:226–43.

15. Zhang D, Ochi N, Takigawa N, Tanimoto Y, Chen Y, Ichihara E, et al. Establishment of pemetrexed-resistant non-small cell lung cancer cell lines. Cancer let. 2011;28:228–35.

16. Mattison LK, Soong R, Diasio RB. Implications of dihydropyrimidine dehydrogenase on 5-fluorouracil pharmacogenetics and pharmacogenomics. Pharmacogenomics. 2002;3:485–92.

17. Diasio RB, Harris BE. Clinical pharmacology of 5-fluorouracil. Clin Pharmacokinet. 1989;16:215–37.

18. Dobritzsch D, Schneider G, Schnackerz KD, Lindqvist Y. Crystal structure of dehydropyrimidine dehydrogenase, a major determinant of the pharmacokinetics of the anti-cancer drug 5-fluorouracil. EMBO J. 2001;208:650–60.

19. Tamura K, Okamoto I, Ozaki T, Kashii T, Takeda K, Kobayashi M, et al. Phase I/II study of S-1 plus carboplatin in patients with advanced non-small cell lung cancer. Eur J Cancer. 2009;45:2132–7.

20. Keira K, Sunaga N, Yanagitani N, Imai H, Utsugi M, Shimizu Y, et al. A phase I dose-escalation study of S-1 plus carboplatin in patients with advanced non-small cell lung cancer. Anticancer Drugs. 2007;18:471–6.

21. Kubota K, Sakai H, Yamamoto N, Kunitoh H, Nakagawa K, Takeda K, et al. A multi-institution phase I/II trial of triweekly regimen with S-1 plus cisplatin in patients with advanced non-small cell lung cancer. J Thorac Oncol. 2010;5:702–6.

22. Lamont EB, Schilsky RL. The oral fluoropyrimidines in cancer chemotherapy. Clin Cancer Res. 1999;5:2289–96.

23. Suehisa H, Toyooka S, Hotta K. Epidermal growth factor receptor mutation status and adjuvant chemotherapy with uracil-tegafur for adenocarcinoma of the lung. J Clin Oncol. 2007;25:3952–7.

24. Mochinaga K, Tsuchiya T, Nagasaki T, Arai J, Tominaga T, Yamasaki N, et al. High expression of dihydropyrimidine dehydrogenase in lung

adenocarcinoma is associated with mutations in epidermal growth factor receptor: implications for the treatment of NSCLC using 5-fluorouracil. Clin Lung Cancer. 2014;15:136–40.

25. Shestopal SA, Johnson MR, Diasio RB. Molecular cloning and characterization of the human dihydropyrimidine dehydrogenase promoter. Biochim Biophys Acta. 2000;1494:162–9.

26. Zhang X, Li L, Fourie J, Davie JR, Guarcello V, Diasio RB, et al. The role of Sp1 and Sp3 in the constitutive DPYD gene expression. Biochim Biophys Acta. 2006;1759:247–56.

27. Lee TC, Oslund KL, Thai P, Velichoko S, Fujisawa T, Duong T, et al. 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin-induced MUC5AC expression: aryl hydrocarbon receptor-independent/EGFR/ERK/p38-dependent SP1-based transcription. Am J Respir Cell Mol Biol. 2011;45:270–6.

28. Blume SW, Snyder RC, Ray R, Thomas S, Koller CA, Miller DM. Mithramycin inhibits SP1 binding and selectively inhibits transcriptional activity of the dehydrofolate reductase gene in vitro and in vivo. J Clin Invest. 1991;88:1613–21.

29. Sordella R, Bell DW, Haber DA, Settleman J. Gefitinib-sensitizing EGFR mutations in lung cancer activate anti-apoptotic pathways. Science. 2004;305:1163–7.

30. Greulich H, Chen TH, Feng W, Janne PA, Alvarez JV, Zappaterra M, et al. Oncogenic transformation by inhibitor-sensitive and -resistant EGFR mutants. ProS Med. 2005;2:1167–76.

31. Marks JL, Gong Y, Chitale D, Golas B, McLellan MD, Kasai Y, et al. Novel MEK1 mutation identified by mutational analysis epidermal growth factor receptor signaling pathway in lung adenocarcinoma. Cancer Res. 2008;68:5524–8.

32. Urick ME, Chung EJ, Shield WP. Enhancement of 5-Fluorouracil-induced in vitro and in vivo radiosensitization with MEK inhibition. Clin Cancer Res. 2011;17:5038–47.