Top

Published in:

01-06-2014 | Original Article

The novel thymidylate synthase inhibitor trifluorothymidine (TFT) and TRAIL synergistically eradicate non-small cell lung cancer cells

Authors: Kaamar Azijli, Ingrid A. M. van Roosmalen, Jorn Smit, Saravanan Pillai, Masakazu Fukushima, Steven de Jong, Godefridus J. Peters, Irene V. Bijnsdorp, Frank A. E. Kruyt

Published in: Cancer Chemotherapy and Pharmacology | Issue 6/2014

Abstract

Purpose

TRAIL, a tumor selective anticancer agent, may be used for the treatment of non-small cell lung cancer (NSCLC). However, TRAIL resistance is frequently encountered. Here, the combined use of TRAIL with trifluorothymidine (TFT), a thymidylate synthase inhibitor, was examined for sensitizing NSCLC cells to TRAIL.

Methods

Interactions between TRAIL and TFT were studied in NSCLC cells using growth inhibition and apoptosis assays. Western blotting and flow cytometry were used to investigate underlying mechanisms.

Results

The combined treatment of TFT and TRAIL showed synergistic cytotoxicity in A549, H292, H322 and H460 cells. For synergistic activity, the sequence of administration was important; TFT treatment followed by TRAIL exposure did not show sensitization. Combined TFT and TRAIL treatment for 24 h followed by 48 h of TFT alone was synergistic in all cell lines, with combination index values below 0.9. The treatments affected cell cycle progression, with TRAIL inducing a G1 arrest and TFT, a G2/M arrest. TFT activated Chk2 and reduced Cdc25c levels known to cause G2/M arrest. TRAIL-induced caspase-dependent apoptosis was enhanced by TFT, whereas TFT alone mainly induced caspase-independent death. TFT increased the expression of p53 and p21/WAF1, and p53 was involved in the increase of TRAIL-R2 surface expression. TFT also caused downregulation of cFLIP and XIAP and increased Bax expression.

Conclusions

TFT enhances TRAIL-induced apoptosis in NSCLC cells by sensitizing the apoptotic machinery at different levels in the TRAIL pathway. Our findings suggest a possible therapeutic benefit of the combined use of TFT and TRAIL in NSCLC.

Jemal A, Siegel R, Ward E, Murray T, Xu J, Thun MJ (2007) Cancer statistics, 2007. CA Cancer J Clin 57(1):43–66PubMedCrossRef

Mountain CF (1995) New prognostic factors in lung cancer. Biologic prophets of cancer cell aggression. Chest 108(1):246–254PubMedCrossRef

Ashkenazi A (2008) Directing cancer cells to self-destruct with pro-apoptotic receptor agonists. Nat Rev Drug Discov 7(12):1001–1012PubMedCrossRef

Wang S, El-Deiry WS (2003) TRAIL and apoptosis induction by TNF-family death receptors. Oncogene 22(53):8628–8633PubMedCrossRef

Danial NN, Korsmeyer SJ (2004) Cell death: critical control points. Cell 116(2):205–219PubMedCrossRef

Voortman J, Resende TP, Abou El Hassan MA, Giaccone G, Kruyt FA (2007) TRAIL therapy in non-small cell lung cancer cells: sensitization to death receptor-mediated apoptosis by proteasome inhibitor bortezomib. Mol Cancer Ther 6(7):2103–2112PubMedCrossRef

Pore MM, Hiltermann TJ, Kruyt FA (2010) Targeting apoptosis pathways in lung cancer. Cancer Lett 332(2):359–368

Bagnoli M, Canevari S, Mezzanzanica D (2010) Cellular FLICE-inhibitory protein (c-FLIP) signalling: a key regulator of receptor-mediated apoptosis in physiologic context and in cancer. Int J Biochem Cell Biol 42(2):210–213PubMedCrossRef

Stegehuis JH, de Wilt LH, de Vries EG, Groen HJ, de Jong S, Kruyt FA (2010) TRAIL receptor targeting therapies for non-small cell lung cancer: current status and perspectives. Drug Resist Updat 13(1–2):2–15PubMedCrossRef

10.

Temmink OH, Emura T, de Bruin M, Fukushima M, Peters GJ (2007) Therapeutic potential of the dual-targeted TAS-102 formulation in the treatment of gastrointestinal malignancies. Cancer Sci 98(6):779–789PubMedCrossRef

11.

Bijnsdorp IV, Peters GJ, Temmink OH, Fukushima M, Kruyt FA (2010) Differential activation of cell death and autophagy results in an increased cytotoxic potential for trifluorothymidine compared to 5-fluorouracil in colon cancer cells. Int J Cancer 126(10):2457–2468PubMed

12.

Broker LE, Kruyt FA, Giaccone G (2005) Cell death independent of caspases: a review. Clin Cancer Res 11(9):3155–3162PubMedCrossRef

13.

Emura T, Murakami Y, Nakagawa F, Fukushima M, Kitazato K (2004) A novel antimetabolite, TAS-102 retains its effect on FU-related resistant cancer cells. Int J Mol Med 13(4):545–549PubMed

14.

Overman MJ, Varadhachary G, Kopetz S, Thomas MB, Fukushima M, Kuwata K, Mita A, Wolff RA, Hoff PM, Xiong H, Abbruzzese JL (2008) Phase 1 study of TAS-102 administered once daily on a 5-day-per-week schedule in patients with solid tumors. Invest New Drugs 26(5):445–454PubMedCrossRef

15.

Yoshino T, Mizunuma N, Yamazaki K, Nishina T, Komatsu Y, Baba H, Tsuji A, Yamaguchi K, Muro K, Sugimoto N, Tsuji Y, Moriwaki T, Esaki T, Hamada C, Tanase T, Ohtsu A (2012) TAS-102 monotherapy for pretreated metastatic colorectal cancer: a double-blind, randomised, placebo-controlled phase 2 trial. Lancet Oncol 13(10):993–1001PubMedCrossRef

16.

Keepers YP, Pizao PE, Peters GJ, van Ark-Otte J, Winograd B, Pinedo HM (1991) Comparison of the sulforhodamine B protein and tetrazolium (MTT) assays for in vitro chemosensitivity testing. Eur J Cancer 27(7):897–900PubMedCrossRef

17.

Bijnsdorp IV, Kruyt FA, Gokoel S, Fukushima M, Peters GJ (2008) Synergistic interaction between trifluorothymidine and docetaxel is sequence dependent. Cancer Sci 99(11):2302–2308PubMedCrossRef

18.

Bijnsdorp IV, Capriotti F, Kruyt FA, Losekoot N, Fukushima M, Griffioen AW, Thijssen VL, Peters GJ (2011) Thymidine phosphorylase in cancer cells stimulates human endothelial cell migration and invasion by the secretion of angiogenic factors. Br J Cancer 104(7):1185–1192PubMedCentralPubMedCrossRef

19.

Mathews ST, Plaisance EP, Kim T (2009) Imaging systems for westerns: chemiluminescence vs. infrared detection. Methods Mol Biol 536:499–513PubMedCrossRef

20.

Koyama S, Koike N, Adachi S (2002) Expression of TNF-related apoptosis-inducing ligand (TRAIL) and its receptors in gastric carcinoma and tumor-infiltrating lymphocytes: a possible mechanism of immune evasion of the tumor. J Cancer Res Clin Oncol 128(2):73–79PubMedCrossRef

21.

Duiker EW, Meijer A, van der Bilt AR, Meersma GJ, Kooi N, van der Zee AG, de Vries EG, de Jong S (2011) Drug-induced caspase 8 upregulation sensitises cisplatin-resistant ovarian carcinoma cells to rhTRAIL-induced apoptosis. Br J Cancer 104(8):1278–1287PubMedCentralPubMedCrossRef

22.

Kastan MB, Bartek J (2004) Cell-cycle checkpoints and cancer. Nature 432(7015):316–323PubMedCrossRef

23.

Mendoza FJ, Ishdorj G, Hu X, Gibson SB (2008) Death receptor-4 (DR4) expression is regulated by transcription factor NF-kappaB in response to etoposide treatment. Apoptosis 13(6):756–770PubMedCrossRef

24.

Frese S, Frese-Schaper M, Andres AC, Miescher D, Zumkehr B, Schmid RA (2006) Cardiac glycosides initiate Apo2L/TRAIL-induced apoptosis in non-small cell lung cancer cells by up-regulation of death receptors 4 and 5. Cancer Res 66(11):5867–5874PubMedCrossRef

25.

Pennarun B, Meijer A, de Vries EG, Kleibeuker JH, Kruyt F, de Jong S (2010) Playing the DISC: turning on TRAIL death receptor-mediated apoptosis in cancer. Biochim Biophys Acta 1805(2):123–140PubMed

26.

Galligan L, Longley DB, McEwan M, Wilson TR, McLaughlin K, Johnston PG (2005) Chemotherapy and TRAIL-mediated colon cancer cell death: the roles of p53, TRAIL receptors, and c-FLIP. Mol Cancer Ther 4(12):2026–2036PubMedCrossRef

27.

Johnston JB, Kabore AF, Strutinsky J, Hu X, Paul JT, Kropp DM, Kuschak B, Begleiter A, Gibson SB (2003) Role of the TRAIL/APO2-L death receptors in chlorambucil- and fludarabine-induced apoptosis in chronic lymphocytic leukemia. Oncogene 22(51):8356–8369PubMedCrossRef

28.

Kitada S, Andersen J, Akar S, Zapata JM, Takayama S, Krajewski S, Wang HG, Zhang X, Bullrich F, Croce CM, Rai K, Hines J, Reed JC (1998) Expression of apoptosis-regulating proteins in chronic lymphocytic leukemia: correlations with In vitro and In vivo chemoresponses. Blood 91(9):3379–3389PubMed

29.

Ewald B, Sampath D, Plunkett W (2008) Nucleoside analogs: molecular mechanisms signaling cell death. Oncogene 27(50):6522–6537PubMedCrossRef

30.

Solier S, Sordet O, Kohn KW, Pommier Y (2009) Death receptor-induced activation of the Chk2- and histone H2AX-associated DNA damage response pathways. Mol Cell Biol 29(1):68–82PubMedCentralPubMedCrossRef

31.

Bosque A, Aguilo JI, del Rey M, Paz-Artal E, Allende LM, Naval J, Anel A (2008) Cell cycle regulation by FasL and Apo2L/TRAIL in human T-cell blasts. Implications for autoimmune lymphoproliferative syndromes. J Leukoc Biol 84(2):488–498PubMedCrossRef

32.

Nagaraj NS, Vigneswaran N, Zacharias W (2006) Cathepsin B mediates TRAIL-induced apoptosis in oral cancer cells. J Cancer Res Clin Oncol 132(3):171–183PubMedCrossRef

33.

Vanden Berghe T, Linkermann A, Jouan-Lanhout S, Walczak H, Vandenabeele P (2014) Regulated necrosis: the expanding network of non-apoptotic cell death pathways. Nat Rev Mol Cell Biol 15(2):135–147PubMedCrossRef

34.

Li LC, Liu GD, Zhang XJ, Li YB (2014) Autophagy, a novel target for chemotherapeutic intervention of thyroid cancer. Cancer Chemother Pharmacol 73(3):439–449PubMedCrossRef

35.

Galvani E, Peters GJ, Giovannetti E (2011) Thymidylate synthase inhibitors for non-small cell lung cancer. Expert Opin Investig Drugs 20(10):1343–1356PubMedCrossRef

36.

Temmink OH, de Bruin M, Comijn EM, Fukushima M, Peters GJ (2005) Determinants of trifluorothymidine sensitivity and metabolism in colon and lung cancer cells. Anticancer Drugs 16(3):285–292PubMedCrossRef

37.

Hong DS, Abbruzzese JL, Bogaard K, Lassere Y, Fukushima M, Mita A, Kuwata K, Hoff PM (2006) Phase I study to determine the safety and pharmacokinetics of oral administration of TAS-102 in patients with solid tumors. Cancer 107(6):1383–1390PubMedCrossRef

38.

Doi T, Ohtsu A, Yoshino T, Boku N, Onozawa Y, Fukutomi A, Hironaka S, Koizumi W, Sasaki T (2012) Phase I study of TAS-102 treatment in Japanese patients with advanced solid tumours. Br J Cancer 107(3):429–434PubMedCentralPubMedCrossRef

39.

Camidge DR, Herbst RS, Gordon MS, Eckhardt SG, Kurzrock R, Durbin B, Ing J, Tohnya TM, Sager J, Ashkenazi A, Bray G, Mendelson D (2010) A phase I safety and pharmacokinetic study of the death receptor 5 agonistic antibody PRO95780 in patients with advanced malignancies. Clin Cancer Res 16(4):1256–1263PubMedCrossRef

40.

Mom CH, Verweij J, Oldenhuis CN, Gietema JA, Fox NL, Miceli R, Eskens FA, Loos WJ, de Vries EG, Sleijfer S (2009) Mapatumumab, a fully human agonistic monoclonal antibody that targets TRAIL-R1, in combination with gemcitabine and cisplatin: a phase I study. Clin Cancer Res 15(17):5584–5590PubMedCrossRef

41.

Soria JC, Mark Z, Zatloukal P, Szima B, Albert I, Juhasz E, Pujol JL, Kozielski J, Baker N, Smethurst D, Hei YJ, Ashkenazi A, Stern H, Amler L, Pan Y, Blackhall F (2011) Randomized phase II study of dulanermin in combination with paclitaxel, carboplatin, and bevacizumab in advanced non-small-cell lung cancer. J Clin Oncol 29(33):4442–4451PubMedCrossRef

Title: The novel thymidylate synthase inhibitor trifluorothymidine (TFT) and TRAIL synergistically eradicate non-small cell lung cancer cells
Authors: Kaamar Azijli
Ingrid A. M. van Roosmalen
Jorn Smit
Saravanan Pillai
Masakazu Fukushima
Steven de Jong
Godefridus J. Peters
Irene V. Bijnsdorp
Frank A. E. Kruyt
Publication date: 01-06-2014
Publisher: Springer Berlin Heidelberg
Published in: Cancer Chemotherapy and Pharmacology / Issue 6/2014
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-014-2465-1

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 6/2014

Impact of variable CYP genotypes on breast cancer relapse in patients undergoing adjuvant tamoxifen therapy

A phase II trial of Xeloda and oxaliplatin (XELOX) neo-adjuvant chemotherapy followed by surgery for advanced gastric cancer patients with para-aortic lymph node metastasis

An open-label study to investigate the cardiac safety profile of cabazitaxel in patients with advanced solid tumors

Phase I study of adjuvant chemotherapy with gemcitabine plus cisplatin in patients with biliary tract cancer undergoing curative resection without major hepatectomy (KHBO1004)

Pharmacokinetics of pazopanib administered in combination with bevacizumab

Mouse pharmacokinetics and metabolism of the curcumin analog, 4-piperidinone,3,5-bis[(2-fluorophenyl)methylene]-acetate(3E,5E) (EF-24; NSC 716993)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer