High pemetrexed sensitivity of docetaxel-resistant A549 cells is mediated by TP53 status and downregulated thymidylate synthase

Kuo,Wei-Ting; Tu,Dom-Gene; Chiu,Ling-Yen; Sheu,Gwo-Tarng; Wu,Ming-Fang

doi:10.3892/or.2017.5951

High pemetrexed sensitivity of docetaxel-resistant A549 cells is mediated by TP53 status and downregulated thymidylate synthase

Authors:
- Wei-Ting Kuo
- Dom-Gene Tu
- Ling-Yen Chiu
- Gwo-Tarng Sheu
- Ming-Fang Wu
View Affiliations

Affiliations: Institute of Clinical Medicine, National Yang-Ming University, Taipei 112, Taiwan, R.O.C., Department of Nuclear Medicine, Ditmanson Medical Foundation, Chia‑Yi Christian Hospital, Chiayi City 60002, Taiwan, R.O.C., Institute of Medicine, Chung Shan Medical University, Taichung City 402, Taiwan, R.O.C., School of Medicine, Chung Shan Medical University, Taichung City 402, Taiwan, R.O.C.
Published online on: September 7, 2017 https://doi.org/10.3892/or.2017.5951
Pages: 2787-2795
Copyright: © Kuo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

The chemoresistance of non-small cell lung cancer (NSCLC) that occurs in docetaxel (DOC) chemotherapy substantially decreases the survival of patients. To overcome DOC-induced chemoresistance, we established DOC-selected A549 lung cancer sublines (A549/D16 and A549/D32) and revealed that both sublines were cross-resistant to vincristine (VCR) and doxorubicin (DXR). Notably, both sublines were more sensitive to pemetrexed (PEM) than parental cells according to MTT and clonogenic assays. The expression levels of thymidylate synthase (TS) and γ-glutamyl hydrolase (GGH) were downregulated in DOC-resistant sublines. When exogenous TS was overexpressed in A549/D16 cells, PEM sensitivity was significantly decreased, however it was not decreased by overexpression of exogenous GGH. PEM treatment induced more apoptotic sub-G1 cells in both DOC-resistant sublines and in the in vivo PEM sensitivities of A549/D16 cells. These findings were further confirmed by a xenografted tumor model. To unmask the mediator of TS downregulation, we investigated human lung cancer cell lines that have various TP53 statuses using DOC treatment. The level of TS protein was significantly decreased in wild-type TP53-containing cells with DOC treatment; TS expression levels were not affected in mutant-TP53 and TP53‑null cells under the same conditions. Furthermore, when the expression of TP53 was inhibited in A549 cells, the expression level of TS was increased. Our data indicated that DOC activated wild-type TP53 and suppressed TS expression under continuous DOC exposure. Therefore, the expression of TS remained at low levels in DOC-resistant A549 cancer cells. Our data revealed that for lung cancer with DOC resistance and wild‑type TP53 status, the administration of PEM as a second-line agent to overcome DOC-resistance may benefit patients.

View Figures

View References

1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Roengvoraphoj M, Tsongalis GJ, Dragnev KH and Rigas JR: Epidermal growth factor receptor tyrosine kinase inhibitors as initial therapy for non-small cell lung cancer: Focus on epidermal growth factor receptor mutation testing and mutation-positive patients. Cancer Treat Rev. 39:839–850. 2013. View Article : Google Scholar : PubMed/NCBI
3	Alamgeer M, Ganju V and Watkins DN: Novel therapeutic targets in non-small cell lung cancer. Curr Opin Pharmacol. 13:394–401. 2013. View Article : Google Scholar : PubMed/NCBI
4	Fitzpatrick FA and Wheeler R: The immunopharmacology of paclitaxel (Taxol), docetaxel (Taxotere), and related agents. Int Immunopharmacol. 3:1699–1714. 2003. View Article : Google Scholar : PubMed/NCBI
5	Horwitz SB: Mechanism of action of taxol. Trends Pharmacol Sci. 13:134–136. 1992. View Article : Google Scholar : PubMed/NCBI
6	Leonard GD, Fojo T and Bates SE: The role of ABC transporters in clinical practice. Oncologist. 8:411–424. 2003. View Article : Google Scholar : PubMed/NCBI
7	Fojo AT and Menefee M: Microtubule targeting agents: Basic mechanisms of multidrug resistance (MDR). Semin Oncol. 32 Suppl 7:S3–S8. 2005. View Article : Google Scholar : PubMed/NCBI
8	Baldwin CM and Perry CM: Pemetrexed: A review of its use in the management of advanced non-squamous non-small cell lung cancer. Drugs. 69:2279–2302. 2009. View Article : Google Scholar : PubMed/NCBI
9	Shih C, Habeck LL, Mendelsohn LG, Chen VJ and Schultz RM: Multiple folate enzyme inhibition: Mechanism of a novel pyrrolopyrimidine-based antifolate LY231514 (MTA). Adv Enzyme Regul. 38:135–152. 1998. View Article : Google Scholar : PubMed/NCBI
10	Goldman ID and Zhao R: Molecular, biochemical, and cellular pharmacology of pemetrexed. Semin Oncol. 29 Suppl 18:S3–S17. 2002. View Article : Google Scholar
11	Zhang D, Ochi N, Takigawa N, Tanimoto Y, Chen Y, Ichihara E, Hotta K, Tabata M, Tanimoto M and Kiura K: Establishment of pemetrexed-resistant non-small cell lung cancer cell lines. Cancer Lett. 309:228–235. 2011. View Article : Google Scholar : PubMed/NCBI
12	Chen CY, Chang YL, Shih JY, Lin JW, Chen KY, Yang CH, Yu CJ and Yang PC: Thymidylate synthase and dihydrofolate reductase expression in non-small cell lung carcinoma: The association with treatment efficacy of pemetrexed. Lung Cancer. 74:132–138. 2011. View Article : Google Scholar : PubMed/NCBI
13	Liu Y, Yin TJ, Zhou R, Zhou S, Fan L and Zhang RG: Expression of thymidylate synthase predicts clinical outcomes of pemetrexed-containing chemotherapy for non-small-cell lung cancer: A systemic review and meta-analysis. Cancer Chemother Pharmacol. 72:1125–1132. 2013. View Article : Google Scholar : PubMed/NCBI
14	Sigmond J, Backus HH, Wouters D, Temmink OH, Jansen G and Peters GJ: Induction of resistance to the multitargeted antifolate Pemetrexed (ALIMTA) in WiDr human colon cancer cells is associated with thymidylate synthase overexpression. Biochem Pharmacol. 66:431–438. 2003. View Article : Google Scholar : PubMed/NCBI
15	Chiu LY, Ko JL, Lee YJ, Yang TY, Tee YT and Sheu GT: L-type calcium channel blockers reverse docetaxel and vincristine-induced multidrug resistance independent of ABCB1 expression in human lung cancer cell lines. Toxicol Lett. 192:408–418. 2010. View Article : Google Scholar : PubMed/NCBI
16	Chiu LY, Hsin IL, Yang TY, Sung WW, Chi JY, Chang JT, Ko JL and Sheu GT: The ERK-ZEB1 pathway mediates epithelial-mesenchymal transition in pemetrexed resistant lung cancer cells with suppression by vinca alkaloids. Oncogene. 36:242–253. 2017. View Article : Google Scholar : PubMed/NCBI
17	Shintani M, Urano M, Takakuwa Y, Kuroda M and Kamoshida S: Immunohistochemical characterization of pyrimidine synthetic enzymes, thymidine kinase-1 and thymidylate synthase, in various types of cancer. Oncol Rep. 23:1345–1350. 2010.PubMed/NCBI
18	Yang TY, Chang GC, Chen KC, Hung HW, Hsu KH, Sheu GT and Hsu SL: Sustained activation of ERK and Cdk2/cyclin-A signaling pathway by pemetrexed leading to S-phase arrest and apoptosis in human non-small cell lung cancer A549 cells. Eur J Pharmacol. 663:17–26. 2011. View Article : Google Scholar : PubMed/NCBI
19	van der Wilt CL, Backus HH, Smid K, Comijn L, Veerman G, Wouters D, Voorn DA, Priest DG, Bunni MA, Mitchell F, et al: Modulation of both endogenous folates and thymidine enhance the therapeutic efficacy of thymidylate synthase inhibitors. Cancer Res. 61:3675–3681. 2001.PubMed/NCBI
20	Chuang JC, Sheu GT, Wang PC, Liao FT, Liu WS, Huang CF, Tseng MH and Wu MF: Docetaxel and 5-fluorouracil induce human p53 tumor suppressor gene transcription via a short sequence at core promoter element. Toxicol In Vitro. 26:678–685. 2012. View Article : Google Scholar : PubMed/NCBI
21	Barak Y, Juven T, Haffner R and Oren M: mdm2 expression is induced by wild type p53 activity. EMBO J. 12:461–468. 1993.PubMed/NCBI
22	Wu X, Bayle JH, Olson D and Levine AJ: The p53-mdm-2 autoregulatory feedback loop. Genes Dev. 7:1126–1132. 1993. View Article : Google Scholar : PubMed/NCBI
23	Chang JT, Chang GC, Ko JL, Liao HY, Liu HJ, Chen CC, Su JM, Lee H and Sheu GT: Induction of tubulin by docetaxel is associated with p53 status in human non small cell lung cancer cell lines. Int J Cancer. 118:317–325. 2006. View Article : Google Scholar : PubMed/NCBI
24	Brown T, Pilkington G, Bagust A, Boland A, Oyee J, Tudur-Smith C, Blundell M, Lai M, Saborido C Martin, Greenhalgh J, et al: Clinical effectiveness and cost-effectiveness of first-line chemotherapy for adult patients with locally advanced or metastatic non-small cell lung cancer: A systematic review and economic evaluation. Health Technol Assess. 17:1–278. 2013. View Article : Google Scholar
25	Yang TY, Chang GC, Chen KC, Hung HW, Hsu KH, Wu CH, Sheu GT and Hsu SL: Pemetrexed induces both intrinsic and extrinsic apoptosis through ataxia telangiectasia mutated/p53-dependent and -independent signaling pathways. Mol Carcinog. 52:183–194. 2013. View Article : Google Scholar : PubMed/NCBI
26	Lee Y, Chen Y, Chang LS and Johnson LF: Inhibition of mouse thymidylate synthase promoter activity by the wild-type p53 tumor suppressor protein. Exp Cell Res. 234:270–276. 1997. View Article : Google Scholar : PubMed/NCBI
27	Calascibetta A, Martorana A, Cabibi D, Aragona F and Sanguedolce R: Relationship between thymidylate synthase and p53 and response to FEC versus taxane adjuvant chemotherapy for breast carcinoma. J Chemother. 23:354–357. 2011. View Article : Google Scholar : PubMed/NCBI
28	Giovannetti E, Backus HH, Wouters D, Ferreira CG, van Houten VM, Brakenhoff RH, Poupon MF, Azzarello A, Pinedo HM and Peters GJ: Changes in the status of p53 affect drug sensitivity to thymidylate synthase (TS) inhibitors by altering TS levels. Br J Cancer. 96:769–775. 2007. View Article : Google Scholar : PubMed/NCBI
29	Horie N and Takeishi K: Identification of functional elements in the promoter region of the human gene for thymidylate synthase and nuclear factors that regulate the expression of the gene. J Biol Chem. 272:18375–18381. 1997. View Article : Google Scholar : PubMed/NCBI
30	Wilson PM, Fazzone W, LaBonte MJ, Lenz HJ and Ladner RD: Regulation of human dUTPase gene expression and p53-mediated transcriptional repression in response to oxaliplatin-induced DNA damage. Nucleic Acids Res. 37:78–95. 2009. View Article : Google Scholar : PubMed/NCBI
31	Bodner SM, Minna JD, Jensen SM, D'Amico D, Carbone D, Mitsudomi T, Fedorko J, Buchhagen DL, Nau MM, Gazdar AF, et al: Expression of mutant p53 proteins in lung cancer correlates with the class of p53 gene mutation. Oncogene. 7:743–749. 1992.PubMed/NCBI
32	Takahashi T, Nau MM, Chiba I, Birrer MJ, Rosenberg RK, Vinocour M, Levitt M, Pass H, Gazdar AF and Minna JD: p53: A frequent target for genetic abnormalities in lung cancer. Science. 246:491–494. 1989. View Article : Google Scholar : PubMed/NCBI