Combined gefitinib and pemetrexed overcome the acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small cell lung cancer

Wu,Min; Yuan,Yuan; Pan,Yue-Yin; Zhang,Ying

doi:10.3892/mmr.2014.2243

Combined gefitinib and pemetrexed overcome the acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small cell lung cancer

Authors:
- Min Wu
- Yuan Yuan
- Yue-Yin Pan
- Ying Zhang
View Affiliations

Affiliations: Department of Geriatrics, The Third Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230061, P.R. China, The Central Laboratory of Binhu Hospital, The Third Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230061, P.R. China, Department of Oncology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, P.R. China
Published online on: May 16, 2014 https://doi.org/10.3892/mmr.2014.2243
Pages: 931-938

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Currently, chemotherapy and targeted therapies share the principal limitation of the emergence of drug resistance, which prevents these strategies from having lasting clinical benefits. The combination of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) with concurrent chemotherapy has been proposed as one strategy to overcome acquired resistance to EGFR-TKIs. The purpose of the present study was to investigate the combined effects of gefitinib and pemetrexed on EGFR-TKI-sensitive and EGFR‑TKI‑resistant human non-small cell lung cancer (NSCLC) cell lines. The antiproliferative effects of gefitinib and pemetrexed, alone and in combination, on the growth of NSCLC cell lines, were assessed using an MTT assay. The cytotoxic interaction between the two drugs was evaluated in vitro using the combination index (CI) method. Cell cycle distribution and apoptosis were analyzed by flow cytometry and alterations in signaling pathways were determined by western blot analysis. In the present study, it was identified that when cells were concurrently exposed to pemetrexed and gefitinib, cytotoxic synergism was present in the gefitinib-resistant PC9/GR human NSCLC cell line and antagonistic interactions were observed in the gefitinib-sensitive PC9 cell line. Synergism was associated with a combination of cell cycle effects of the different agents. In addition, the combination of pemetrexed and gefitinib decreased the levels of phosphorylated AKT, phosphorylated extracellular-signal-regulated kinase and B-cell lymphoma 2 as compared with those in the control. By contrast, antagonism was associated with gefitinib-induced G0/G1-phase blockade of gefitinib-sensitive cells, which interfered with the cell cycle-specific cytotoxicity of chemotherapy. The combination of pemetrexed and gefitinib generated synergistic effects in gefitinib-acquired resistant cells and antagonistic effects in gefitinib-sensitive cells, suggesting that EGFR-TKIs combined with pemetrexed may be a beneficial treatment strategy for NSCLC patients with acquired resistance to EGFR-TKIs.

View Figures

View References

1	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar
2	Salomon DS, Brandt R, Ciardiello F and Normanno N: Epidermal growth factor-related peptides and their receptors in human malignancies. Crit Rev Oncol Hematol. 19:183–232. 1995. View Article : Google Scholar : PubMed/NCBI
3	Ennis BW, Lippman ME and Dickson RB: The EGF receptor system as a target for antitumor therapy. Cancer Invest. 9:553–562. 1991. View Article : Google Scholar : PubMed/NCBI
4	Lynch TJ, Bell DW, Sordella R, et al: Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med. 350:2129–2139. 2004. View Article : Google Scholar : PubMed/NCBI
5	Maemondo M, Inoue A, Kobayashi K, et al: Gefitinib or chemotherapy for non-small-cell-lung cancer with mutated EGFR. N Engl J Med. 362:2380–2388. 2010. View Article : Google Scholar : PubMed/NCBI
6	Masago K, Fujita S, Togashi Y, et al: Clinicopathologic factors affecting the progression free survival of patients with advanced non-small-cell lung cancer after gefitinib therapy. Clin Lung Cancer. 12:56–61. 2011. View Article : Google Scholar : PubMed/NCBI
7	Jackman D, Pao W, Riely GJ, et al: Clinical definition of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small-cell lung cancer. J Clin Oncol. 28:357–360. 2010. View Article : Google Scholar : PubMed/NCBI
8	Lovly CM and Horn L: Strategies for overcoming EGFR resistance in the treatment of advanced-stage NSCLC. Curr Treat Options Oncol. 13:516–526. 2012. View Article : Google Scholar : PubMed/NCBI
9	Goldberg SB, Oxnard GR, Digumarthy S, et al: Chemotherapy with erlotinib or chemotherapy alone in advanced non-small cell lung cancer with acquired resistance to EGFR tyrosine kinase inhibitors. Oncologist. 18:1214–1220. 2013. View Article : Google Scholar : PubMed/NCBI
10	Haddad R, Allen A, Wirth L, Tishler R and Posner M: Integrating novel agents into the curative treatment of head and neck cancer. Expert Rev Anticancer Ther. 6:157–159. 2006. View Article : Google Scholar : PubMed/NCBI
11	Hanna N, Shepherd FA, Fossella FV, et al: Randomized phase III trial of pemetrexed versus docetaxel in patients with non-small cell lung cancer previously treated with chemotherapy. J Clin Oncol. 22:1589–1597. 2004. View Article : Google Scholar : PubMed/NCBI
12	Fukuoka M, Yano S, Giaccone G, et al: Multi-institutional randomized phase II trial of gefitinib for previously treated patients with advanced non-small-cell lung cancer (The IDEAL 1 Trial) [corrected]. J Clin Oncol. 21:2237–2246. 2003.
13	Grunwald V and Hidalgo M: Developing inhibitors of the epidermal growth factor receptor for cancer treatment. J Natl Cancer Inst. 95:851–867. 2003. View Article : Google Scholar : PubMed/NCBI
14	Kim ES, Hirsh V, Mok T, et al: Gefitinib versus docetaxel in previously treated non-small-cell lung cancer (INTEREST): a randomised phase III trial. Lancet. 372:1809–1818. 2008. View Article : Google Scholar : PubMed/NCBI
15	Steins MB, Reinmuth N, Bischoff H, Kindermann M and Thomas M: Targeting the epidermal growth factor receptor in non-small cell lung cancer. Onkologie. 33:704–709. 2010. View Article : Google Scholar : PubMed/NCBI
16	Baselga J and Arteaga CL: Critical update and emerging trends in epidermal growth factor receptor targeting in cancer. J Clin Oncol. 23:2445–2459. 2005. View Article : Google Scholar : PubMed/NCBI
17	Herbst RS and Sandler AB: Overview of the current status of human epidermal growth factor receptor inhibitors in lung cancer. Clin Lung Cancer. 6(Suppl 1): S7–S19. 2004. View Article : Google Scholar : PubMed/NCBI
18	Ciardiello F, Caputo R, Borriello G, et al: ZD1839 (IRESSA), an EGFR-selective tyrosine kinase inhibitor, enhances taxane activity in bcl-2 overexpressing, multidrug-resistant MCF-7 ADR human breast cancer cells. Int J Cancer. 98:463–469. 2002. View Article : Google Scholar
19	Park JK, Lee SH, Kang JH, Nishio K, Saijo N and Kuh HJ: Synergistic interaction between gefitinib (Iressa, ZD1839) and paclitaxel against human gastric carcinoma cells. Anticancer Drugs. 15:809–818. 2004. View Article : Google Scholar
20	Sirotnak FM, Zakowski MF, Miller VA, Scher HI and Kris MG: Efficacy of cytotoxic agents against human tumor xenografts is markedly enhanced by coadministration of ZD1839 (Iressa), an inhibitor of EGFR tyrosine kinase. Clin Cancer Res. 6:4885–4892. 2000.PubMed/NCBI
21	Shih C, Chen VJ, Gossett LS, et al: LY231514, a pyrrolo[2,3-d]pyrimidine-based antifolate that inhibits multiple folate-requiring enzymes. Cancer Res. 57:1116–1123. 1997.
22	Chattopadhyay S, Moran RG and Goldman ID: Pemetrexed: biochemical and cellular pharmacology, mechanisms, and clinical applications. Mol Cancer Ther. 6:404–417. 2007. View Article : Google Scholar : PubMed/NCBI
23	Scagliotti GV, Parikh P, von Pawel J, et al: Phase III study comparing cisplatin plus gemcitabine with cisplatin plus pemetrexed in chemotherapy-naive patients with advanced-stage non-small-cell lung cancer. J Clin Oncol. 26:3543–3551. 2008. View Article : Google Scholar : PubMed/NCBI
24	Ciuleanu T, Brodowicz T, Zielinski C, et al: Maintenance pemetrexed plus best supportive care versus placebo plus best supportive care for non-small-cell lung cancer: a randomised, double-blind, phase 3 study. Lancet. 374:1432–1440. 2009. View Article : Google Scholar : PubMed/NCBI
25	Chou TC and Talalay P: Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul. 22:27–55. 1984. View Article : Google Scholar : PubMed/NCBI
26	Rinaldi DA, Kuhn JG, Burris HA, et al: A phase I evaluation of multitargeted antifolate (MTA, LY231514), administered every 21 days, utilizing the modified continual reassessment method for dose escalation. Cancer Chemother Pharmacol. 44:372–380. 1999. View Article : Google Scholar
27	Thödtmann R, Depenbrock H, Dumez H, et al: Clinical and pharmacokinetic phase I study of multitargeted antifolate (LY231514) in combination with cisplatin. J Clin Oncol. 17:3009–3016. 1999.PubMed/NCBI
28	D’Addario G, Pintilie M, Leighl NB, Feld R, Cerny T and Shepherd FA: Platinum-based versus non-platinum-based chemotherapy in advanced non-small-cell lung cancer: a meta-analysis of the published literature. J Clin Oncol. 23:2926–2936. 2005.PubMed/NCBI
29	Pfister DG, Johnson DH, Azzoli CG, et al: American Society of Clinical Oncology treatment of unresectable non-small-cell lung cancer guideline: update 2003. J Clin Oncol. 22:330–353. 2004. View Article : Google Scholar : PubMed/NCBI
30	Ardizzoni A and Tiseo M: Second-line chemotherapy in the treatment of advanced non-small cell lung cancer (NSCLC). J Chemother. 16(Suppl 4): 104–107. 2004. View Article : Google Scholar : PubMed/NCBI
31	Maemondo M: Timing the change of chemotherapy for non small cell lung cancer. Gan To Kagaku Ryoho. 39:1316–1319. 2012.(In Japanese).
32	Oxnard GR, Arcila ME, Chmielecki J, Ladanyi M, Miller VA and Pao W: New strategies in overcoming acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in lung cancer. Clin Cancer Res. 17:5530–5537. 2011. View Article : Google Scholar : PubMed/NCBI
33	Van Schaeybroeck S, Kyula J, Kelly DM, et al: Chemotherapy-induced epidermal growth factor receptor activation determines response to combined gefitinib/chemotherapy treatment in non-small cell lung cancer cells. Mol Cancer Ther. 5:1154–1165. 2006.
34	Li T, Ling YH, Goldman ID and Perez-Soler R: Schedule-dependent cytotoxic synergism of pemetrexed and erlotinib in human non-small cell lung cancer cells. Clin Cancer Res. 13:3413–3422. 2007. View Article : Google Scholar : PubMed/NCBI
35	Giovannetti E, Lemos C, Tekle C, et al: Molecular mechanisms underlying the synergistic interaction of erlotinib, an epidermal growth factor receptor tyrosine kinase inhibitor, with the multitargeted antifolate pemetrexed in non-small-cell lung cancer cells. Mol Pharmacol. 73:1290–1300. 2008. View Article : Google Scholar
36	Davies AM, Ho C, Lara PN Jr, Mack P, Gumerlock PH and Gandara DR: Pharmacodynamic separation of epidermal growth factor receptor tyrosine kinase inhibitors and chemotherapy in non-small-cell lung cancer. Clin Lung Cancer. 7:385–388. 2006. View Article : Google Scholar : PubMed/NCBI
37	Ruvolo PP, Deng X and May WS: Phosphorylation of Bcl-2 and regulation of apoptosis. Leukemia. 15:515–522. 2001. View Article : Google Scholar : PubMed/NCBI
38	Diel P, Smolnikar K and Michna H: The pure antiestrogen ICI 182780 is more effective in the induction of apoptosis and down regulation of BCL-2 than tamoxifen in MCF-7 cells. Breast Cancer Res Treat. 58:87–97. 1999. View Article : Google Scholar : PubMed/NCBI
39	Sordella R, Bell DW, Haber DA and Settleman J: Gefitinib-sensitizing EGFR mutations in lung cancer activate anti-apoptotic pathways. Science. 305:1163–1167. 2004. View Article : Google Scholar : PubMed/NCBI
40	Amann J, Kalyankrishna S, Massion PP, et al: Aberrant epidermal growth factor receptor signaling and enhanced sensitivity to EGFR inhibitors in lung cancer. Cancer Res. 65:226–235. 2005.PubMed/NCBI
41	Van Schaeybroeck S, Karaiskou-McCaul A, Kelly D, et al: EGFR activity determines response of colorectal cancer cells to gefitinib (Iressa) alone and in combination with chemotherapy. Clin Cancer Res. 11:7480–7489. 2005.PubMed/NCBI