Skip to main content

Advertisement

SpringerLink
Log in

Arsenic induces apoptosis in myoblasts through a reactive oxygen species-induced endoplasmic reticulum stress and mitochondrial dysfunction pathway

  • Inorganic Compounds
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

A pool of myoblasts available for myogenesis is important for skeletal muscle size. The decreased number of skeletal muscle fibers could be due to the decreased myoblast proliferation or cytotoxicity. Identification of toxicants that regulate myoblast apoptosis is important in skeletal muscle development or regeneration. Here, we investigate the cytotoxic effect and its possible mechanisms of arsenic trioxide (As2O3) on myoblasts. C2C12 myoblasts underwent apoptosis in response to As2O3 (1–10 μM), accompanied by increased Bax/Bcl-2 ratio, decreased mitochondria membrane potential, increased cytochrome c release, increased caspase-3/-9 activity, and increased poly (ADP-ribose) polymerase (PARP) cleavage. Moreover, As2O3 triggered the endoplasmic reticulum (ER) stress indentified through several key molecules of the unfolded protein response, including glucose-regulated protein (GRP)-78, GRP-94, PERK, eIF2α, ATF6, and caspase-12. Pretreatment with antioxidant N-acetylcysteine (NAC, 0.5 mM) dramatically suppressed the increases in reactive oxygen species (ROS), lipid peroxidation, ER stress, caspase cascade activity, and apoptosis in As2O3 (10 μM)-treated myoblasts. Furthermore, As2O3 (10 μM) effectively decreased the phosphorylation of Akt, which could be reversed by NAC. Over-expression of constitutive activation of Akt (c.a. Akt) also significantly attenuated As2O3-induced myoblast apoptosis. Taken together, these results suggest that As2O3 may exert its cytotoxicity on myoblasts by inducing apoptosis through a ROS-induced mitochondrial dysfunction, ER stress, and Akt inactivation signaling pathway.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Adams JM, Cory S (2007) The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 26:1324–1337

    Article  CAS  PubMed  Google Scholar 

  • Antonsson B (2001) Bax and other pro-apoptotic Bcl-2 family “killer-proteins” and their victim the mitochondrion. Cell Tissue Res 306:347–361

    Article  CAS  PubMed  Google Scholar 

  • Boonstra J, Post JA (2004) Molecular events associated with reactive oxygen species and cell cycle progression in mammalian cells. Gene 337:1–13

    Article  CAS  PubMed  Google Scholar 

  • Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ, Arden KC, Blenis J, Greenberg ME (1999) Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 96:857–868

    Article  CAS  PubMed  Google Scholar 

  • Cai BZ, Meng FY, Zhu SL, Zhao J, Liu JQ, Liu CJ, Chen N, Ye ML, Li ZY, Ai J, Lu YJ, Yang BF (2010) Arsenic trioxide induces the apoptosis in bone marrow mesenchymal stem cells by intracellular calcium signal and caspase-3 pathways. Toxicol Lett 193:173–178

    Article  CAS  PubMed  Google Scholar 

  • Chao DT, Korsmeyer SJ (1998) BCL-2 family: regulators of cell death. Annu Rev Immunol 16:395–419

    Article  CAS  PubMed  Google Scholar 

  • Charge SB, Rudnicki MA (2004) Cellular and molecular regulation of muscle regeneration. Physiol Rev 84:209–238

    Article  CAS  PubMed  Google Scholar 

  • Chen YC, Lin-Shiau SY, Lin JK (1998) Involvement of reactive oxygen species and caspase 3 activation in arsenite-induced apoptosis. J Cell Physiol 177:324–333

    Article  CAS  PubMed  Google Scholar 

  • Chen YW, Huang CF, Tsai KS, Yang RS, Yen CC, Yang CY, Lin-Shiau SY, Liu SH (2006) Methylmercury induces pancreatic beta-cell apoptosis and dysfunction. Chem Res Toxicol 19:1080–1085

    Article  CAS  PubMed  Google Scholar 

  • Chen L, Xu B, Liu L, Luo Y, Yin J, Zhou H, Chen W, Shen T, Han X, Huang S (2010) Hydrogen peroxide inhibits mTOR signaling by activation of AMPKalpha leading to apoptosis of neuronal cells. Lab Invest 90:762–773

    Article  CAS  PubMed  Google Scholar 

  • Choi YJ, Park JW, Suh SI, Mun KC, Bae JH, Song DK, Kim SP, Kwon TK (2002) Arsenic trioxide-induced apoptosis in U937 cells involve generation of reactive oxygen species and inhibition of Akt. Int J Oncol 21:603–610

    CAS  PubMed  Google Scholar 

  • Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y, Greenberg ME (1997) Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91:231–241

    Article  CAS  PubMed  Google Scholar 

  • Deponti D, Francois S, Baesso S, Sciorati C, Innocenzi A, Broccoli V, Muscatelli F, Meneveri R, Clementi E, Cossu G, Brunelli S (2007) Necdin mediates skeletal muscle regeneration by promoting myoblast survival and differentiation. J Cell Biol 179:305–319

    Article  CAS  PubMed  Google Scholar 

  • Florea AM, Splettstoesser F, Busselberg D (2007) Arsenic trioxide (As2O3) induced calcium signals and cytotoxicity in two human cell lines: SY-5Y neuroblastoma and 293 embryonic kidney (HEK). Toxicol Appl Pharmacol 220:292–301

    Article  CAS  PubMed  Google Scholar 

  • Fresno Vara JA, Casado E, de Castro J, Cejas P, Belda-Iniesta C, Gonzalez-Baron M (2004) PI3K/Akt signalling pathway and cancer. Cancer Treat Rev 30:193–204

    Article  PubMed  Google Scholar 

  • Garrido C, Galluzzi L, Brunet M, Puig PE, Didelot C, Kroemer G (2006) Mechanisms of cytochrome c release from mitochondria. Cell Death Differ 13:1423–1433

    Article  CAS  PubMed  Google Scholar 

  • Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312

    Article  CAS  PubMed  Google Scholar 

  • Hawke TJ, Garry DJ (2001) Myogenic satellite cells: physiology to molecular biology. J Appl Physiol 91:534–551

    CAS  PubMed  Google Scholar 

  • Hockenbery D, Nunez G, Milliman C, Schreiber RD, Korsmeyer SJ (1990) Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature 348:334–336

    Article  CAS  PubMed  Google Scholar 

  • Jansen KM, Pavlath GK (2008) Prostaglandin F2alpha promotes muscle cell survival and growth through upregulation of the inhibitor of apoptosis protein BRUCE. Cell Death Differ 15:1619–1628

    Article  CAS  PubMed  Google Scholar 

  • Jiang B, Xiao W, Shi Y, Liu M, Xiao X (2005a) Heat shock pretreatment inhibited the release of Smac/DIABLO from mitochondria and apoptosis induced by hydrogen peroxide in cardiomyocytes and C2C12 myogenic cells. Cell Stress Chaperones 10:252–262

    Article  CAS  PubMed  Google Scholar 

  • Jiang B, Xiao W, Shi Y, Liu M, Xiao X (2005b) Role of Smac/DIABLO in hydrogen peroxide-induced apoptosis in C2C12 myogenic cells. Free Radic Biol Med 39:658–667

    Article  CAS  PubMed  Google Scholar 

  • Jimi S, Uchiyama M, Takaki A, Suzumiya J, Hara S (2004) Mechanisms of cell death induced by cadmium and arsenic. Ann NY Acad Sci 1011:325–331

    Article  CAS  PubMed  Google Scholar 

  • Kaufman RJ (1999) Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls. Genes Dev 13:1211–1233

    Article  CAS  PubMed  Google Scholar 

  • Kuo ML, Chuang SE, Lin MT, Yang SY (2001) The involvement of PI 3-K/Akt-dependent up-regulation of Mcl-1 in the prevention of apoptosis of Hep3B cells by interleukin-6. Oncogene 20:677–685

    Article  CAS  PubMed  Google Scholar 

  • Lazebnik YA, Kaufmann SH, Desnoyers S, Poirier GG, Earnshaw WC (1994) Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature 371:346–347

    Article  CAS  PubMed  Google Scholar 

  • Li JX, Shen YQ, Cai BZ, Zhao J, Bai X, Lu YJ, Li XQ (2010) Arsenic trioxide induces the apoptosis in vascular smooth muscle cells via increasing intracellular calcium and ROS formation. Mol Biol Rep 37:1569–1576

    Article  CAS  PubMed  Google Scholar 

  • Lu TH, Su CC, Chen YW, Yang CY, Wu CC, Hung DZ, Chen CH, Cheng PW, Liu SH, Huang CF (2011) Arsenic induces pancreatic beta-cell apoptosis via the oxidative stress-regulated mitochondria-dependent and endoplasmic reticulum stress-triggered signaling pathways. Toxicol Lett 201:15–26

    Article  CAS  PubMed  Google Scholar 

  • Ma Y, Brewer JW, Diehl JA, Hendershot LM (2002) Two distinct stress signaling pathways converge upon the CHOP promoter during the mammalian unfolded protein response. J Mol Biol 318:1351–1365

    Article  CAS  PubMed  Google Scholar 

  • Manning BD, Cantley LC (2007) AKT/PKB signaling: navigating downstream. Cell 129:1261–1274

    Article  CAS  PubMed  Google Scholar 

  • Mauro A (1961) Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol 9:493–495

    Article  CAS  PubMed  Google Scholar 

  • Mitchell PO, Pavlath GK (2001) A muscle precursor cell-dependent pathway contributes to muscle growth after atrophy. Am J Physiol Cell Physiol 281:C1706–C1715

    CAS  PubMed  Google Scholar 

  • Miyake M, Hayashi S, Iwasaki S, Uchida T, Watanabe K, Ohwada S, Aso H, Yamaguchi T (2011) TIEG1 negatively controls the myoblast pool indispensable for fusion during myogenic differentiation of C2C12 cells. J Cell Physiol 226:1128–1136

    Article  CAS  PubMed  Google Scholar 

  • Morishima N, Nakanishi K, Takenouchi H, Shibata T, Yasuhiko Y (2002) An endoplasmic reticulum stress-specific caspase cascade in apoptosis. Cytochrome c-independent activation of caspase-9 by caspase-12. J Biol Chem 277:34287–34294

    Article  CAS  PubMed  Google Scholar 

  • Nakagawa T, Yuan J (2000) Cross-talk between two cysteine protease families. Activation of caspase-12 by calpain in apoptosis. J Cell Biol 150:887–894

    Article  CAS  PubMed  Google Scholar 

  • Navas-Acien A, Sharrett AR, Silbergeld EK, Schwartz BS, Nachman KE, Burke TA, Guallar E (2005) Arsenic exposure and cardiovascular disease: a systematic review of the epidemiologic evidence. Am J Epidemiol 162:1037–1049

    Article  PubMed  Google Scholar 

  • Orrenius S, Gogvadze V, Zhivotovsky B (2007) Mitochondrial oxidative stress: implications for cell death. Annu Rev Pharmacol Toxicol 47:143–183

    Article  CAS  PubMed  Google Scholar 

  • Osaki M, Oshimura M, Ito H (2004) PI3K-Akt pathway: its functions and alterations in human cancer. Apoptosis 9:667–676

    Article  CAS  PubMed  Google Scholar 

  • Oyadomari S, Mori M (2004) Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 11:381–389

    Article  CAS  PubMed  Google Scholar 

  • Porter AG, Janicke RU (1999) Emerging roles of caspase-3 in apoptosis. Cell Death Differ 6:99–104

    Article  CAS  PubMed  Google Scholar 

  • Qi XJ, Wildey GM, Howe PH (2006) Evidence that Ser87 of BimEL is phosphorylated by Akt and regulates BimEL apoptotic function. J Biol Chem 281:813–823

    Article  CAS  PubMed  Google Scholar 

  • Raghu KG, Cherian OL (2009) Characterization of cytotoxicity induced by arsenic trioxide (a potent anti-APL drug) in rat cardiac myocytes. J Trace Elem Med Biol 23:61–68

    Article  CAS  PubMed  Google Scholar 

  • Reed JC (1995) Regulation of apoptosis by bcl-2 family proteins and its role in cancer and chemoresistance. Curr Opin Oncol 7:541–546

    Article  CAS  PubMed  Google Scholar 

  • Ron D, Walter P (2007) Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 8:519–529

    Article  CAS  PubMed  Google Scholar 

  • Rosenblatt JD, Yong D, Parry DJ (1994) Satellite cell activity is required for hypertrophy of overloaded adult rat muscle. Muscle Nerve 17:608–613

    Article  CAS  PubMed  Google Scholar 

  • Shen ZX, Chen GQ, Ni JH, Li XS, Xiong SM, Qiu QY, Zhu J, Tang W, Sun GL, Yang KQ, Chen Y, Zhou L, Fang ZW, Wang YT, Ma J, Zhang P, Zhang TD, Chen SJ, Chen Z, Wang ZY (1997) Use of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients. Blood 89:3354–3360

    CAS  PubMed  Google Scholar 

  • Steffens AA, Hong GM, Bain LJ (2011) Sodium arsenite delays the differentiation of C2C12 mouse myoblast cells and alters methylation patterns on the transcription factor myogenin. Toxicol Appl Pharmacol 250:154–161

    Article  CAS  PubMed  Google Scholar 

  • Szegezdi E, Fitzgerald U, Samali A (2003) Caspase-12 and ER-stress-mediated apoptosis: the story so far. Ann NY Acad Sci 1010:186–194

    Article  CAS  PubMed  Google Scholar 

  • Szegezdi E, Logue SE, Gorman AM, Samali A (2006) Mediators of endoplasmic reticulum stress-induced apoptosis. EMBO Rep 7:880–885

    Article  CAS  PubMed  Google Scholar 

  • Tang CH, Chiu YC, Huang CF, Chen YW, Chen PC (2009) Arsenic induces cell apoptosis in cultured osteoblasts through endoplasmic reticulum stress. Toxicol Appl Pharmacol 241:173–181

    Article  CAS  PubMed  Google Scholar 

  • Tews DS, Goebel HH (1997) DNA-fragmentation and expression of apoptosis-related proteins in muscular dystrophies. Neuropathol Appl Neurobiol 23:331–338

    Article  CAS  PubMed  Google Scholar 

  • Tidball JG, Albrecht DE, Lokensgard BE, Spencer MJ (1995) Apoptosis precedes necrosis of dystrophin-deficient muscle. J Cell Sci 108:2197–2204

    CAS  PubMed  Google Scholar 

  • Todd DJ, Lee AH, Glimcher LH (2008) The endoplasmic reticulum stress response in immunity and autoimmunity. Nat Rev Immunol 8:663–674

    Article  CAS  PubMed  Google Scholar 

  • Tophkhane C, Yang S, Bales W, Archer L, Osunkoya A, Thor AD, Yang X (2007) Bcl-2 overexpression sensitizes MCF-7 cells to genistein by multiple mechanisms. Int J Oncol 31:867–874

    CAS  PubMed  Google Scholar 

  • Torres M, Forman HJ (2003) Redox signaling and the MAP kinase pathways. BioFactors 17:287–296

    Article  CAS  PubMed  Google Scholar 

  • Wang XZ, Harding HP, Zhang Y, Jolicoeur EM, Kuroda M, Ron D (1998) Cloning of mammalian Ire1 reveals diversity in the ER stress responses. EMBO J 17:5708–5717

    Article  CAS  PubMed  Google Scholar 

  • Yadav S, Shi Y, Wang F, Wang H (2010) Arsenite induces apoptosis in human mesenchymal stem cells by altering Bcl-2 family proteins and by activating intrinsic pathway. Toxicol Appl Pharmacol 244:263–272

    Article  CAS  PubMed  Google Scholar 

  • Yen YP, Tsai KS, Chen YW, Huang CF, Yang RS, Liu SH (2010) Arsenic inhibits myogenic differentiation and muscle regeneration. Environ Health Perspect 118:949–956

    Article  CAS  PubMed  Google Scholar 

  • Yen CC, Ho TJ, Wu CC, Chang CF, Su CC, Chen YW, Jinn TR, Lu TH, Cheng PW, Su YC, Liu SH, Huang CF (2011) Inorganic arsenic causes cell apoptosis in mouse cerebrum through an oxidative stress-regulated signaling pathway. Arch Toxicol 85:565–575

    Article  CAS  PubMed  Google Scholar 

  • Yoshida H, Okada T, Haze K, Yanagi H, Yura T, Negishi M, Mori K (2000) ATF6 activated by proteolysis binds in the presence of NF-Y (CBF) directly to the cis-acting element responsible for the mammalian unfolded protein response. Mol Cell Biol 20:6755–6767

    Article  CAS  PubMed  Google Scholar 

  • Yoshida T, Yamauchi H, Fan Sun G (2004) Chronic health effects in people exposed to arsenic via the drinking water: dose-response relationships in review. Toxicol Appl Pharmacol 198:243–252

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Duncan G, Wang L, Liu P, Cui H, Reddan JR, Yang BF, Wormstone IM (2007) Arsenic trioxide initiates ER stress responses, perturbs calcium signalling and promotes apoptosis in human lens epithelial cells. Exp Eye Res 85:825–835

    Article  CAS  PubMed  Google Scholar 

  • Zou H, Yang R, Hao J, Wang J, Sun C, Fesik SW, Wu JC, Tomaselli KJ, Armstrong RC (2003) Regulation of the Apaf-1/caspase-9 apoptosome by caspase-3 and XIAP. J Biol Chem 278:8091–8098

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by grant from the National Science Council of Taiwan (NSC97-2314-B-002-052-MY3).

Conflict of interest

The authors declare no conflict of interest.

Author information

Authors and Affiliations

  1. College of Medicine, Institute of Toxicology, National Taiwan University, Taipei, Taiwan

    Yuan-Peng Yen & Shing-Hwa Liu

  2. Department of Laboratory Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan

    Keh-Sung Tsai

  3. Department of Physiology, China Medical University, Taichung, Taiwan

    Ya-Wen Chen

  4. Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan

    Ya-Wen Chen

  5. School of Chinese Medicine, China Medical University, Taichung, Taiwan

    Chun-Fa Huang

  6. Department of Orthopaedic, Collegelege of Medicine and Hospital, National Taiwan University, Taipei, Taiwan

    Rong-Sen Yang

  7. Department of Urology, College of Medicine and Hospital, National Taiwan University, Taipei, Taiwan

    Shing-Hwa Liu

Authors
  1. Yuan-Peng Yen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Keh-Sung Tsai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ya-Wen Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chun-Fa Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rong-Sen Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shing-Hwa Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Rong-Sen Yang or Shing-Hwa Liu.

Additional information

Yuan-Peng Yen and Keh-Sung Tsai contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yen, YP., Tsai, KS., Chen, YW. et al. Arsenic induces apoptosis in myoblasts through a reactive oxygen species-induced endoplasmic reticulum stress and mitochondrial dysfunction pathway. Arch Toxicol 86, 923–933 (2012). https://doi.org/10.1007/s00204-012-0864-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-012-0864-9

Keywords

Search

Navigation