Paraquat Induces Apoptosis through Cytochrome C Release and ERK Activation

  • Seo, Hong Joo (Department of Thoracic and Cardiovascular Surgery, Chosun University Hospital) ;
  • Choi, Sang Joon (Department of Obstetrics and Gynecology, Chosun University Hospital) ;
  • Lee, Jung-Hee (Department of Cellular and Molecular Medicine, Chosun University School of Medicine)
  • Received : 2014.10.14
  • Accepted : 2014.11.04
  • Published : 2014.11.30


Paraquat has been suggested to induce apoptosis by generation of reactive oxygen species (ROS). However, little is known about the mechanism of paraquat-induced apoptosis. Here, we demonstrate that extracellular signal-regulated protein kinase (ERK) is required for paraquat-induced apoptosis in NIH3T3 cells. Paraquat treatment resulted in activation of ERK, and U0126, inhibitors of the MEK/ERK signaling pathway, prevented apoptosis. Moreover, paraquat-induced apoptosis was associated with cytochrome C release, which could be prevented by treatment with the MEK inhibitors. Taken together, our findings suggest that ERK activation plays an active role in mediating paraquat-induced apoptosis of NIH3T3 cells.



Supported by : Ministry of Education, Science and Technology


  1. Agrawal, M., Bhaskar, A. S. and Rao, P. V. (2014) Involvement of mitogen-activated protein kinase pathway in T-2 toxin-induced cell cycle alteration and apoptosis in human neuroblastoma cells. Mol. Neurobiol. Aug 2. [Epub ahead of print]
  2. Bost, F., McKay, R., Dean, N. and Mercola, D. (1997) The JUN kinase/stress-activated protein kinase pathway is required for epidermal growth factor stimulation of growth of human A549 lung carcinoma cells. J. Biol. Chem. 272, 33422-33429.
  3. Chang, X., Lu, W., Dou, T., Wang, X., Lou, D., Sun, X. and Zhou, Z. (2013) Paraquat inhibits cell viability via enhanced oxidative stress and apoptosis in human neural progenitor cells. Chem. Biol. Interact. 206, 248-255.
  4. Craig, E. A., Stevens, M. V., Vaillancourt, R. R. and Camenisch, T. D. (2008) MAP3Ks as central regulators of cell fate during development. Dev. Dyn. 237, 3102-3114.
  5. Darling, N. J. and Cook, S. J. (2014) The role of MAPK signalling pathways in the response to endoplasmic reticulum stress. Biochim. Biophys. Acta 1843, 2150-2163.
  6. Deschenes-Simard, X., Kottakis, F., Meloche, S. and Ferbeyre, G. (2014) ERKs in cancer: friends or foes? Cancer Res. 74, 412-419.
  7. Ding, H. D., Zhang, X. H., Xu, S. C., Sun, L. L., Jiang, M. Y., Zhang, A. Y. and Jin, Y. G. (2009) Induction of protection against paraquat-induced oxidative damage by abscisic acid in maize leaves is mediated through mitogen-activated protein kinase. J. Integr. Plant Biol. 51, 961-972.
  8. Esmaeili, M. A., Farimani, M. M. and Kiaei, M. (2014) Anticancer effect of calycopterin via PI3K/Akt and MAPK signaling pathways, ROSmediated pathway and mitochondrial dysfunction in hepatoblastoma cancer (HepG2) cells. Mol. Cell. Biochem. 397, 17-31
  9. Fei, Q. and Ethell, D. W. (2008) Maneb potentiates paraquat neurotoxicity by inducing key Bcl-2 family members. J. Neurochem. 105, 2091-2097.
  10. Fei, Q., McCormack, A. L., Di Monte, D. A. and Ethell, D. W. (2008) Paraquat neurotoxicity is mediated by a Bak-dependent mechanism. J. Biol. Chem. 283, 3357-3364.
  11. Giordano, S., Darley-Usmar, V. and Zhang, J. (2014) Autophagy as an essential cellular antioxidant pathway in neurodegenerative disease. Redox Biol. 2, 82-90.
  12. Gold, M. R. (2008) B cell development: important work for ERK. Immunity 28, 488-490.
  13. Haberzettl, P. and Hill, B. G. (2013) Oxidized lipids activate autophagy in a JNK-dependent manner by stimulating the endoplasmic reticulum stress response. Redox Biol. 1, 56-64.
  14. Han, J., Zhang, Z., Yang, S., Wang, J., Yang, X. and Tan, D. (2014) Betanin attenuates paraquat-induced liver toxicity through a mitochondrial pathway. Food Chem. Toxicol. 70, 100-106.
  15. Hong, G. L., Liu, J. M., Zhao, G. J., Wang, L., Liang, G., Wu, B., Li, M. F., Qiu, Q. M. and Lu, Z. Q. (2013) The reversal of paraquatinduced mitochondria-mediated apoptosis by cycloartenyl ferulate, the important role of Nrf2 pathway. Exp. Cell Res. 319, 2845-2855.
  16. Huttemann, M., Pecina, P., Rainbolt, M., Sanderson, T. H., Kagan, V. E., Samavati, L., Doan, J. W. and Lee, I. (2011) The multiple functions of cytochrome c and their regulation in life and death decisions of the mammalian cell: From respiration to apoptosis. Mitochondrion 11, 369-381.
  17. Kim, D., Koo, J. S. and Lee, S. (2014) Overexpression of reactive oxygen species scavenger enzymes is associated with a good prognosis in triple-negative breast cancer. Oncology 88, 9-17.
  18. Kim, E. K. and Choi, E. J. (2010) Pathological roles of MAPK signaling pathways in human diseases. Biochim. Biophys. Acta 1802, 396-405.
  19. Kumar, P., Rao, G. N., Pal, B. B. and Pal, A. (2014) Hyperglycemia-induced oxidative stress induces apoptosis by inhibiting PI3-kinase/ Akt and ERK1/2 MAPK mediated signaling pathway causing downregulation of 8-oxoG-DNA glycosylase levels in glial cells. Int. J. Biochem. Cell Biol. 53, 302-319.
  20. Landes, T. and Martinou, J. C. (2011) Mitochondrial outer membrane permeabilization during apoptosis: the role of mitochondrial fission. Biochim. Biophys. Acta 1813, 540-545.
  21. Lavrik, I. N. and Krammer, P. H. (2012) Regulation of CD95/Fas signaling at the DISC. Cell Death Differ. 19, 36-41.
  22. Li, J., O, W., Li, W., Jiang, Z. G. and Ghanbari, H. A. (2013) Oxidative stress and neurodegenerative disorders. Int. J. Mol. Sci. 14, 24438-24475.
  23. Lim, N. R., Thomas, C. J., Silva, L. S., Yeap, Y. Y., Yap, S., Bell, J. R., Delbridge, L. M., Bogoyevitch, M. A., Woodman, O. L., Williams, S. J., May, C. N. and Ng, D. C. (2013) Cardioprotective 3',4'-dihydroxyflavonol attenuation of JNK and p38(MAPK) signalling involves CaMKII inhibition. Biochem. J. 456, 149-161.
  24. Liu, P., Kong, F., Wang, J., Xu, H., Qi, T. and Meng, J. (2014) Involvement of IGF-1 and MEOX2 in PI3K/Akt1/2 and ERK1/2 pathways mediated proliferation and differentiation of perivascular adipocytes. Exp. Cell. Res. Sep 18. pii: S0014-4827 (14)00400-5. doi: 10.1016/j.yexcr.2014.09.011. [Epub ahead of print]
  25. Mak, S. K., Tewari, D., Tetrud, J. W., Langston, J. W. and Schule, B. (2011) Mitochondrial dysfunction in skin fibroblasts from a Parkinson's disease patient with an alpha-synuclein triplication. J. Parkinsons Dis. 1, 175-183.
  26. Meierjohann, S. (2014) Oxidative stress in melanocyte senescence and melanoma transformation. Eur. J .Cell Biol. 93, 36-41.
  27. Miller, R. L., Sun, G. Y. and Sun, A. Y. (2007) Cytotoxicity of paraquat in microglial cells: Involvement of PKCdelta- and ERK1/2-dependent NADPH oxidase. Brain Res. 1167, 129-139.
  28. Monian, P. and Jiang, X. (2012) Clearing the final hurdles to mitochondrial apoptosis: regulation post cytochrome C release. Exp. Oncol. 34, 185-191.
  29. Munshi, A. and Ramesh, R. (2013) Mitogen-activated protein kinases and their role in radiation response. Genes Cancer 4, 401-408.
  30. Nahirnyj, A., Livne-Bar, I., Guo, X. and Sivak, J. M. (2013) ROS detoxification and proinflammatory cytokines are linked by p38 MAPK signaling in a model of mature astrocyte activation. PLoS One 8, e83049.
  31. Nikoletopoulou, V., Markaki, M., Palikaras, K. and Tavernarakis, N. (2013) Crosstalk between apoptosis, necrosis and autophagy. Biochim. Biophys. Acta 1833, 3448-3459.
  32. Ohmichi, M., Hayakawa, J., Tasaka, K., Kurachi, H. and Murata, Y. (2005) Mechanisms of platinum drug resistance. Trends Pharmacol. Sci. 26, 113-116.
  33. Prakash, J., Yadav, S. K., Chouhan, S. and Singh, S. P. (2013) Neuroprotective role of Withania somnifera root extract in maneb-paraquat induced mouse model of parkinsonism. Neurochem. Res. 38, 972-980.
  34. Qin, X., Wu, Q., Lin, L., Sun, A., Liu, S., Li, X., Cao, X., Gao, T., Luo, P., Zhu, X. and Wang, X. (2014) Soluble epoxide hydrolase deficiency or inhibition attenuates MPTP-induced parkinsonism. Mol. Neurobiol. Aug 17. [Epub ahead of print]
  35. Rincheval, V., Bergeaud, M., Mathieu, L., Leroy, J., Guillaume, A., Mignotte, B., Le Floch, N. and Vayssiere, J. L. (2012) Differential effects of Bcl-2 and caspases on mitochondrial permeabilization during endogenous or exogenous reactive oxygen species-induced cell death: a comparative study of H(2)O(2), paraquat, t-BHP, etoposide and TNF-alpha-induced cell death. Cell Biol.Toxicol. 28, 239-253.
  36. Son, Y., Kim, S., Chung, H. T. and Pae, H. O. (2013) Reactive oxygen species in the activation of MAP kinases. Methods Enzymol. 528, 27-48.
  37. Tian, F., Dong, L., Zhou, Y., Shao, Y., Li, W., Zhang, H. and Wang, F. (2014a) Rapamycin-Induced apoptosis in HGF-stimulated lens epithelial cells by AKT/mTOR, ERK and JAK2/STAT3 pathways. Int. J. Mol. Sci. 15, 13833-13848.
  38. Tian, H., Zhang, D., Gao, Z., Li, H., Zhang, B., Zhang, Q., Li, L., Cheng, Q., Pei, D. and Zheng, J. (2014b) MDA-7/IL-24 inhibits Nrf2-mediated antioxidant response through activation of p38 pathway and inhibition of ERK pathway involved in cancer cell apoptosis. Cancer Gene Ther. Sep 19. doi: 10.1038/cgt.2014.45. [Epub ahead of print]
  39. Tormos, A. M., Talens-Visconti, R., Nebreda, A. R. and Sastre, J. (2013) p38 MAPK: a dual role in hepatocyte proliferation through reactive oxygen species. Free Radic. Res. 47, 905-916.
  40. Wallace, M. A., Bailey, S., Fukuto, J. M., Valentine, J. S. and Gralla, E. B. (2005) Induction of phenotypes resembling CuZn-superoxide dismutase deletion in wild-type yeast cells: an in vivo assay for the role of superoxide in the toxicity of redox-cycling compounds. Chem. Res. Toxicol. 18, 1279-1286.
  41. Wang, F., Franco, R., Skotak, M., Hu, G. and Chandra, N. (2014a) Mechanical stretch exacerbates the cell death in SH-SY5Y cells exposed to paraquat: mitochondrial dysfunction and oxidative stress. Neurotoxicology 41, 54-63.
  42. Wang, J., Deng, X., Zhang, F., Chen, D. and Ding, W. (2014b) ZnO nanoparticle-induced oxidative stress triggers apoptosis by activating JNK signaling pathway in cultured primary astrocytes. Nanoscale Res. Lett. 9, 117.
  43. Wang, X., Luo, F. and Zhao, H. (2014c) Paraquat-induced reactive oxygen species inhibit neutrophil apoptosis via a p38 MAPK/NFkappaB-IL-6/TNF-alpha positive-feedback circuit. PLoS One 9, e93837.

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