The online version of this article (doi:10.1186/1476-4598-11-25) contains supplementary material, which is available to authorized users.
Aitziber Buqué, Jangi Sh Muhialdin contributed equally to this work.
The authors declare that they have no competing interests.
AB undertook the experimental procedures as well as preparing and writing the manuscript. JSM undertook the apoptosis experiments as well as preparing and writing the manuscript. AB and JSM contributed equally to this work. UA and BC undertook the gene expression assays. UA, SC and AM did the statistical analysis for the manuscript. SC, AM, AS and IR proposed the chemotherapy for the study. GLV was central in the design, coordination and manuscript editing. All authors have read and approved the final manuscript.
Metastatic melanoma is a lethal skin cancer and its incidence is rising every year. It represents a challenge for oncologist, as the current treatment options are non-curative in the majority of cases; therefore, the effort to find and/or develop novel compounds is mandatory. Pemetrexed (Alimta®, MTA) is a multitarget antifolate that inhibits folate-dependent enzymes: thymidylate synthase, dihydrofolate reductase and glycinamide ribonucleotide formyltransferase, required for de novo synthesis of nucleotides for DNA replication. It is currently used in the treatment of mesothelioma and non-small cell lung cancer (NSCLC), and has shown clinical activity in other tumors such as breast, colorectal, bladder, cervical, gastric and pancreatic cancer. However, its effect in human melanoma has not been studied yet.
In the current work we studied the effect of MTA on four human melanoma cell lines A375, Hs294T, HT144 and MeWo and in two NSCLC cell lines H1299 and Calu-3. We have found that MTA induces DNA damage, S-phase cell cycle arrest, and caspase- dependent and –independent apoptosis. We show that an increment of the intracellular reactive oxygen species (ROS) and p53 is required for MTA-induced cytotoxicity by utilizing N-Acetyl-L-Cysteine (NAC) to blockage of ROS and p53-defective H1299 NSCLC cell line. Pretreatment of melanoma cells with NAC significantly decreased the DNA damage, p53 up-regulation and cytotoxic effect of MTA. MTA was able to induce p53 expression leading to up-regulation of p53-dependent genes Mcl-1 and PIDD, followed by a postranscriptional regulation of Mcl-1 improving apoptosis.
We found that MTA induced DNA damage and mitochondrial-mediated apoptosis in human melanoma cells in vitro and that the associated apoptosis was both caspase-dependent and –independent and p53-mediated. Our data suggest that MTA may be of therapeutic relevance for the future treatment of human malignant melanoma.
Authors’ original file for figure 112943_2011_1045_MOESM1_ESM.pdf
Authors’ original file for figure 212943_2011_1045_MOESM2_ESM.pdf
Authors’ original file for figure 312943_2011_1045_MOESM3_ESM.pdf
Authors’ original file for figure 412943_2011_1045_MOESM4_ESM.pdf
Authors’ original file for figure 512943_2011_1045_MOESM5_ESM.tiff
Authors’ original file for figure 612943_2011_1045_MOESM6_ESM.pdf
Authors’ original file for figure 712943_2011_1045_MOESM7_ESM.ppt
American Cancer Society: What Are the Key Statistics About Melanoma?. [ http://www.cancer.org/Cancer/SkinCancer-Melanoma/DetailedGuide/melanoma-skin-cancer-key-statistics], 
National Cancer Institute: Technical Notes for the SEER Cancer Statistics Review. 2011, 1975-2008.
Jangi SM, az-Perez JL, Ochoa-Lizarralde B, Martin-Ruiz I, Asumendi A, Perez-Yarza G, Gardeazabal J, az-Ramon JL, Boyano MD: H1 Histamine Receptor Antagonists Induce Genotoxic and Caspase-2-Dependent Apoptosis in Human Melanoma Cells. Carcinogenesis. 2006, 27: 1787-1796. 10.1093/carcin/bgl021 CrossRefPubMed
Zamzami N, Kroemer G: Methods to Measure Membrane Potential and Permeability Transition in the Mitochondria During Apoptosis. Methods Mol Biol. 2004, 282: 103-115. PubMed
Ramsby ML, Makowski GS, Khairallah EA: Differential Detergent Fractionation of Isolated Hepatocytes: Biochemical, Immunochemical and Two-Dimensional Gel Electrophoresis Characterization of Cytoskeletal and Noncytoskeletal Compartments. Electrophoresis. 1994, 15: 265-277. 10.1002/elps.1150150146 CrossRefPubMed
Kim JM, Bae HR, Park BS, Lee JM, Ahn HB, Rho JH, Yoo KW, Park WC, Rho SH, Yoon HS, Yoo YH: Early Mitochondrial Hyperpolarization and Intracellular Alkalinization in Lactacystin-Induced Apoptosis of Retinal Pigment Epithelial Cells. J Pharmacol Exp Ther. 2003, 305: 474-481. 10.1124/jpet.102.047811 CrossRefPubMed
Norman P: Pemetrexed Disodium (Eli Lilly). Curr Opin Investig Drugs. 2001, 2: 1611-1622. PubMed
Teicher BA, Chen V, Shih C, Menon K, Forler PA, Phares VG, Amsrud T: Treatment Regimes Including the Multitargeted Antifolate LY231514 in Human Tumor Xenografts. Clin Cancer Res. 2000, 6: 1016-1023. PubMed
Balch CM, Buzaid AC, Soong SJ, Atkins MB, Cascinelli N, Coit DG, Fleming ID, Gershenwald JE, Houghton A, Kirkwood JM, McMasters KM, Mihm MF, Morton DL, Reintgen DS, Ross MI, Sober A, Thompson JA, Thompson JF: Final Version of the American Joint Committee on Cancer Staging System for Cutaneous Melanoma. J Clin Oncol. 2001, 19: 3635-3648. PubMed
Manola J, Atkins M, Ibrahim J, Kirkwood J: Prognostic Factors in Metastatic Melanoma: a Pooled Analysis of Eastern Cooperative Oncology Group Trials. J Clin Oncol. 2000, 18: 3782-3793. PubMed
Ramirez JM, San Miguel JF, Ocio EM, Pandiella A: Pemetrexed Acts As an Antimyeloma Agent by Provoking Cell Cycle Blockade and Apoptosis. Leukaemia. 2007, 21: 797-804.
Slee EA, Harte MT, Kluck RM, Wolf BB, Casiano CA, Newmeyer DD, Wang HG, Reed JC, Nicholson DW, Alnemri ES, Green DR, Martin SJ: Ordering the Cytochrome C-Initiated Caspase Cascade: Hierarchical Activation of Caspases-2, -3, -6, -7, -8, and −10 in a Caspase-9-Dependent Manner. J Cell Biol. 1999, 144: 281-292. 10.1083/jcb.144.2.281 PubMedCentralCrossRefPubMed
Ruefli AA, Ausserlechner MJ, Bernhard D, Sutton VR, Tainton KM, Kofler R, Smyth MJ, Johnstone RW: The Histone Deacetylase Inhibitor and Chemotherapeutic Agent Suberoylanilide Hydroxamic Acid (SAHA) Induces a Cell-Death Pathway Characterized by Cleavage of Bid and Production of Reactive Oxygen Species. Proc Natl Acad Sci U S A. 2001, 98: 10833-10838. 10.1073/pnas.191208598 PubMedCentralCrossRefPubMed
Calin GA, Cimmino A, Fabbri M, Ferracin M, Wojcik SE, Shimizu M, Taccioli C, Zanesi N, Garzon R, Aqeilan RI, Alder H, Volinia S, Rassenti L, Liu X, Liu CG, Kipps TJ, Negrini M, Croce CM: MiR-15a and MiR-16-1 Cluster Functions in Human Leukemia. Proc Natl Acad Sci U S A. 2008, 105: 5166-5171. 10.1073/pnas.0800121105 PubMedCentralCrossRefPubMed
Thallinger C, Wolschek MF, Wacheck V, Maierhofer H, Gunsberg P, Polterauer P, Pehamberger H, Monia BP, Selzer E, Wolff K, Jansen B: Mcl-1 Antisense Therapy Chemosensitizes Human Melanoma in a SCID Mouse Xenotransplantation Model. J Invest Dermatol. 2003, 120: 1081-1086. 10.1046/j.1523-1747.2003.12252.x CrossRefPubMed
- Molecular mechanism implicated in Pemetrexed-induced apoptosis in human melanoma cells
Jangi Sh Muhialdin
- BioMed Central
Neu im Fachgebiet Onkologie
e.Med Kampagnen-Visual, Mail Icon II