Pemetrexed Induces G1 Phase Arrest and Apoptosis through Inhibiting Akt Activation in Human Non Small Lung Cancer Cell Line A549

  • Wu, Dong-Ming (State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University) ;
  • Zhang, Peng (Department of Radiation Oncology, Sichuan Cancer Hospital) ;
  • Xu, Guang-Chao (State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University) ;
  • Tong, Ai-Ping (State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University) ;
  • Zhou, Cong (State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University) ;
  • Lang, Jin-Yi (Department of Radiation Oncology, Sichuan Cancer Hospital) ;
  • Wang, Chun-Ting (State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University)
  • Published : 2015.03.09


Pemetrexed is an antifolate agent which has been used for treating malignant pleural mesothelioma and non small lung cancer in the clinic as a chemotherapeutic agent. In this study, pemetrexed inhibited cell growth and induced G1 phase arrest in the A549 cell line. To explore the molecular mechanisms of pemetrexed involved in cell growth, we used a two-dimensional polyacrylamide gel electrophoresis (2-DE) proteomics approach to analyze proteins changed in A549 cells treated with pemetrexed. As a result, twenty differentially expressed proteins were identified by ESI-Q-TOF MS/MS analysis in A549 cells incubated with pemetrexed compared with non-treated A549 cells. Three key proteins (GAPDH, HSPB1 and EIF4E) changed in pemetrexed treated A549 cells were validated by Western blotting. Accumulation of GAPDH and decrease of HSPB1 and EIF4E which induce apoptosis through inhibiting phosphorylation of Akt were noted. Expression of p-Akt in A549 cells treated with pemetrexed was reduced. Thus, pemetrexed induced apoptosis in A549 cells through inhibiting the Akt pathway.


Pemetrexed;2-DE;Akt;apoptosis;A549 non small cell lung cancer cells


Supported by : National Natural Science Foundation of China


  1. Adjei AA (2000). Pemetrexed: a multitargeted antifolate agent with promising activity in solid tumors. Ann Oncol, 11, 1335-41.
  2. Adjei AA (2004). Pemetrexed (ALIMTA), a novel multitargeted antineoplastic agent. Clin Cancer Res, 10, 4276-80.
  3. Bareschino MA, Schettino C, Rossi A, et al (2011). Treatment of advanced non small cell lung cancer. J Thoracic Disease, 3, 122-33.
  4. Buque A, Muhialdin JS, Munoz A, et al (2012). Molecular mechanism implicated in Pemetrexedinduced apoptosis in human melanoma cells. Mol Cancer, 11, 25.
  5. Cohen MH, Johnson JR, Wang YC, Sridhara R, Pazdur R (2005). FDA drug approval summary: pemetrexed for injection (Alimta) for the treatment of non-small cell lung cancer. Oncologist, 10, 363-8.
  6. Colell A, Green DR, Ricci JE (2009). Novel roles for GAPDH in cell death and carcinogenesis. Cell Death Differ, 16, 1573-81.
  7. Dai H, Chen Y, Elmquist WF (2005). Distribution of the novel antifolate pemetrexed to the brain. J Pharmacol Exp Ther, 315, 222-9.
  8. De Benedetti A, Graff JR (2004). eIF-4E expression and its role in malignancies and metastases. Oncogene, 23, 3189-99.
  9. De Boer RH, Arrieta O, Yang CH, et al (2011). Vandetanib plus pemetrexed for the second-line treatment of advanced nonsmall-cell lung cancer: a randomized, double-blind phase III trial. J Clin Oncol, 29, 1067-74.
  10. Deng BG, Yao JH, Liu QY, et al (2013). Comparative serum proteomic analysis of serum diagnosis proteins of colorectal cancer based on magnetic bead separation and maldi-tof mass spectrometry. Asian Pac J Cancer Prev, 14, 6069-75.
  11. Ferlay J, Shin HR, Bray F, et al (2010). Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer, 127, 2893-917.
  12. Gettinger S, Lynch T (2011). A decade of advances in treatment for advanced non-small cell lung cancer. Clin Chest Med, 32, 839-51.
  13. Hayashi N, Peacock JW, Beraldi E, et al (2012). Hsp27 silencing coordinately inhibits proliferation and promotes Fas-induced apoptosis by regulating the PEA-15 molecular switch. Cell Death Differ, 19, 990-1002.
  14. Molina JR, Adjei AA (2003). The role of Pemetrexed (Alimta, LY231514) in lung cancer therapy. Clin Lung Cancer, 5, 21-7.
  15. Leisner TM, Moran C, Holly SP, Parise LV (2012). CIB1 prevents nuclear GAPDH accumulation and non-apoptotic tumor cell death via AKT and ERK signaling. Oncogene.
  16. Li XJ, Wu QF, He DL, Fu JK, Jin X (2013). Proteomic profiling of serum from stage I lung squamous cell carcinoma patients. Asian Pac J Cancer Prev, 14, 2273-6.
  17. Luo X, Lamsal LP, Xu WJ, et al (2014). Genetic variant in CLPTM1L confers reduced risk of lung cancer: a replication study in Chinese and a meta-analysis. Asian Pac J Cancer Prev, 15, 9241-7.
  18. Parisa K, Armin S, Mohammad RR (2014). Implementation of proteomics for cancer research: past, present, and future. Asian Pac J Cancer Prev, 15, 2433-38.
  19. Ramirez JM, Ocio EM, San Miguel JF, Pandiella A (2007). Pemetrexed acts as an antimyeloma agent by provoking cell cycle blockade and apoptosis. Leukemia, 21, 797-804.
  20. Rane MJ, Pan Y, Singh S, et al (2003). Heat shock protein 27 controls apoptosis by regulating Akt activation. J Biol Chem, 278, 27828-35.
  21. Raizer JJ, Rademaker A, Evens AM, et al (2012). Pemetrexed in the treatment of relapsed/refractory primary central nervous system lymphoma. Cancer, 118, 3743-8.
  22. Rothbart SB, Racanelli AC, Moran RG (2010). Pemetrexed indirectly activates the metabolic kinase AMPK in human carcinomas. Cancer Res, 70, 10299-309.
  23. Smit EF, Socinski MA, Mullaney BP, et al (2012). Biomarker analysis in a phase III study of pemetrexed-carboplatin versus etoposide-carboplatin in chemonaive patients with extensive-stage small-cell lung cancer. Ann Oncol, 23, 1723-9.
  24. Smith PG, Marshman E, Newell DR, Curtin NJ (2000). Dipyridamole potentiates the in vitro activity of MTA (LY231514) by inhibition of thymidine transport. Br J Cancer, 82, 924-30.
  25. Socinski MA, Smit EF, Lorigan P, et al (2009). Phase III study of pemetrexed plus carboplatin compared with etoposide plus carboplatin in chemotherapy-naive patients with extensivestage small-cell lung cancer. J Clin Oncol, 27, 4787-92.
  26. Sun SY, Rosenberg LM, Wang X, et al (2005). Activation of Akt and eIF4E survival pathways by rapamycin-mediated mammalian target of rapamycin inhibition. Cancer Res, 65, 7052-8.
  27. Stinchcombe TE, Socinski MA (2009). Current treatments for advanced stage non-small cell lung cancerc. Proc Am Thoracic Soc, 6, 233-41.
  28. Tarze A, Deniaud A, Le Bras M, et al (2007). GAPDH, a novel regulator of the pro-apoptotic mitochondrial membrane permeabilization. Oncogene, 26, 2606-20.
  29. Tong A, Wu L, Lin Q, et al (2008). Proteomic analysis of cellular protein alterations using a hepatitis B virus-producing cellular model. Proteomics, 8, 2012-23
  30. Tristan C, Shahani N, Sedlak TW, Sawa A (2011). The diverse functions of GAPDH: views from different subcellular compartments. Cell Signal, 23, 317-23
  31. Wendel HG, De Stanchina E, Fridman JS, et al (2004). Survival signalling by Akt and eIF4E in oncogenesis and cancer therapy. Nature, 428, 332-7
  32. Wu MF, Hsiao YM, Huang CF, et al (2010). Genetic determinants of pemetrexed responsiveness and nonresponsiveness in non-small cell lung cancer cells. J Thorac Oncol, 5, 1143-51.
  33. Wu R, Kausar H, Johnson P, et al (2007). Hsp27 regulates Akt activation and polymorphonuclear leukocyte apoptosis by scaffolding MK2 to Akt signal complex. J Biol Chem, 282, 21598-608.
  34. Yang TY, Chang GC, Chen KC, et al (2011). Pemetrexed induces both intrinsic and extrinsic apoptosis through ataxia telangiectasia mutated/p53-dependent and -independent signaling pathways. Mol Carcinogen.
  35. Yang TY, Chang GC, Chen KC, et al (2011). 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.
  36. Yu Z, Chen XZ, Cui LH, et al (2014). Prediction of lung cancer based on serum biomarkers by gene expression programming methods. Asian Pac J Cancer Prev, 15, 9367-73.

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