The online version of this article (doi:10.1186/1476-4598-11-34) contains supplementary material, which is available to authorized users.
The authors declare that they have no competing interests.
LT synthesized the compound IK11 and made original observations leading to this work. GF Jr and SB designed the work and GF Jr contributed to the critical revision of the script. EP conducted cell cycle analyses, RB measured apoptosis and necrosis and mitochondrial depolarization with flow cytometry. VB and TZ accomplished fluorescent microscopy, AC, PJK performed immunoblot analysis and TZ made the wound healing assay. RB contributed to the conception and design of the entire study, performed the rest of the experiments and wrote the initial drafts of the manuscript. All authors read and approved the final manuscript.
2,4-Dimethoxyphenyl-E-4-arylidene-3-isochromanone (IK11) was previously described to induce apoptotic death of A431 tumor cells. In this report, we investigated the molecular action of IK11 in the HepG2 human hepatocellular carcinoma cell line to increase our knowledge of the role of poly (ADP-ribose)-polymerase (PARP), protein kinase B/Akt and mitogen activated protein kinase (MAPK) activation in the survival and death of tumor cells and to highlight the possible role of PARP-inhibitors in co-treatments with different cytotoxic agents in cancer therapy.
We found that sublethal concentrations of IK11 prevented proliferation, migration and entry of the cells into their G2 phase. At higher concentrations, IK11 induced reactive oxygen species (ROS) production, mitochondrial membrane depolarization, activation of c-Jun N-terminal kinase 2 (JNK2), and substantial loss of HepG2 cells. ROS production appeared marginal in mediating the cytotoxicity of IK11 since N-acetyl cysteine was unable to prevent it. However, the PARP inhibitor PJ34, although not a ROS scavenger, strongly inhibited both IK11-induced ROS production and cell death. JNK2 activation seemed to be a major mediator of the effect of IK11 since inhibition of JNK resulted in a substantial cytoprotection while inhibitors of the other kinases failed to do so. Inhibition of Akt slightly diminished the effect of IK11, while the JNK and Akt inhibitor and ROS scavenger trans-resveratrol completely protected against it.
These results indicate significant involvement of PARP, a marginal role of ROS and a pro-apoptotic role of Akt in this system, and raise attention to a novel mechanism that should be considered when cancer therapy is augmented with PARP-inhibition, namely the cytoprotection by inhibition of JNK2.
Authors’ original file for figure 112943_2011_1029_MOESM1_ESM.tiff
Authors’ original file for figure 212943_2011_1029_MOESM2_ESM.tiff
Authors’ original file for figure 312943_2011_1029_MOESM3_ESM.tiff
Authors’ original file for figure 412943_2011_1029_MOESM4_ESM.tiff
Authors’ original file for figure 512943_2011_1029_MOESM5_ESM.tiff
Authors’ original file for figure 612943_2011_1029_MOESM6_ESM.tiff
Veres B, Gallyas F, Varbiro G, Berente Z, Osz E, Szekeres G, Szabo C, Sumegi B: Decrease of the inflammatory response and induction of the Akt/protein kinase B pathway by poly-(ADP-ribose) polymerase 1 inhibitor in endotoxin-induced septic shock. Biochem Pharmacol. 2003, 65 (8): 1373-1382. 10.1016/S0006-2952(03)00077-7 CrossRefPubMed
Szanto A, Bognar Z, Szigeti A, Szabo A, Farkas L, Gallyas F: Critical Role of Bad Phosphorylation by Akt in Cytostatic Resistance of Human Bladder Cancer Cells. Anticancer Res. 2009, 29: 159-164. PubMed
Mester L, Szabo A, Atlasz T, Szabadfi K, Reglodi D, Kiss P, Racz B, Tamas A, Gallyas F, Sumegi B, Hocsak E, Gabriel R, Kovacs K: Protection Against Chronic Hypoperfusion-Induced Retinal Neurodegeneration by PARP Inhibition via Activation of PI-3-kinase Akt Pathway and Suppression of JNK and p38 MAP Kinases. Neurotox Res. 2009, 16 (1): 68-76. 10.1007/s12640-009-9049-6 CrossRefPubMed
Abdallah Y, Gligorievski D, Kasseckert SA, Dieterich L, Schäfer M, Kuhlmann CR, Noll T, Sauer H, Piper HM, Schäfer C: The role of poly(ADP-ribose) polymerase (PARP) in the autonomous proliferative response of endothelial cells to hypoxia. Cardiovasc Res. 2007, 1 (3): 568-574. 73 CrossRef
Clark JS, Faisal A, Baliga R, Nagamine Y, Arany I: Cisplatin induces apoptosis through the ERK-p66shc pathway in renal proximal tubule cells. Cancer Lett. 2010, 28 (2): 165-170. 297 CrossRef
Racz B, Hanto K, Tapodi A, Solti I, Kalman N, Jakus P, Kovacs K, Debreceni B, Gallyas F, Sumegi B: Regulation of MKP-1 expression and MAPK activation by PARP-1 in oxidative stress: A new mechanism for cytoplasmic effect of PARP-1 activation. Free Radic Biol Med. 2010, 49 (12): 1978-1988. 10.1016/j.freeradbiomed.2010.09.026 CrossRefPubMed
Tapodi A, Debreceni B, Hanto K, Bognar Z, Wittmann I, Gallyas F, Varbiro G, Sumegi B: Pivotal role of Akt activation in mitochondrial protection and cell survival by poly(ADP-ribose)polymerase-1 inhibition in oxidative stress. J Biol Chem. 2005, 280 (42): 35767-35775. 10.1074/jbc.M507075200 CrossRefPubMed
Lemasters JJ, Nieminen AL, Qian T, Trost LC, Elmore SP, Nishimura Y, Crowe RA, Cascio WE, Bradham CA, Brenner DA, Herman B: The mitochondrial permeability transition in cell death: a common mechanism in necrosis, apoptosis and autophagy. Biochim Biophys Acta. 1998, 1366: 177-196. 10.1016/S0005-2728(98)00112-1 CrossRefPubMed
Lorand T, Forgo P, Foldesi A, Osz E, Prokai L: Improved solvent-free synthesis and structure elucidation of (E)- and (Z)-4-arylmethylene-3-isochromanones. Eur J Org Chem. 2002, 17: 2996-3003. CrossRef
Kalai T, Balog M, Szabo A, Gulyas G, Jeko J, Sumegi B, Hideg K: New poly(ADP-ribose) polymerase-1 inhibitors with antioxidant activity based on 4-carboxamidobenzimidazole-2-ylpyrroline and -tetrahydropyridine nitroxides and their precursors. J Med Chem. 2009, 52 (6): 1619-1629. 10.1021/jm801476y CrossRefPubMed
Palfi A, Toth A, Kulcsar G, Hanto K, Deres P, Bartha E, Halmosi R, Szabados E, Czopf L, Kalai T, Hideg K, Sumegi B, Toth K: The role of Akt and mitogen-activated protein kinase systems in the protective effect of poly-(ADP-ribose)-polymerase inhibition in Langendorff perfused and in isoproterenol-damaged rat hearts. J Pharmacol Exp Ther. 2005, 315 (1): 273-282. 10.1124/jpet.105.088336 CrossRefPubMed
Bartha E, Solti I, Kereskai L, Lantos J, Plozer E, Magyar K, Szabados E, Kálai T, Hideg K, Halmosi R, Sumegi B, Toth K: PARP inhibition delays transition of hypertensive cardiopathy to heart failure in spontaneously hypertensive rats. Cardiovasc Res. 2009, 83 (3): 501-510. 10.1093/cvr/cvp144 CrossRefPubMed
Sykes SM, Lane SW, Bullinger L, Kalaitzidis D, Yusuf R, Saez B, Ferraro F, Mercier F, Singh H, Brumme KM, Acharya SS, Schöll C, Tothova Z, Attar EC, Fröhling S, DePinho RA, Armstrong SA, Gilliland DG, Scadden DT: AKT/FOXO Signaling Enforces Reversible Differentiation Blockade in Myeloid Leukemias. Cell. 2011, 146 (5): 697-708. 10.1016/j.cell.2011.07.032 CrossRefPubMed
Kaul R, Saha P, Saradhi M, Prasad LA, Chatterjee S, Ghosh I, Tyagi RK, Datta K: Overexpression of hyaluronan binding protein 1 (HABP1/p32/gC1qR) in HepG2 cell leads to increased hyaluronan synthesis and cell proliferation by upregulation of cyclin D1 in AKT-dependent pathway. J Biol Chem. 2012, [Epub ahead of print]
- Protective effect of the poly(ADP-ribose) polymerase inhibitor PJ34 on mitochondrial depolarization-mediated cell death in hepatocellular carcinoma cells involves attenuation of c-Jun N-terminal kinase-2 and protein kinase B/Akt activation
Janos Krisztian Priber
Ferenc Gallyas Jr
- BioMed Central
Neu im Fachgebiet Onkologie
Mail Icon II