• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. Jun 1, 2000; 348(Pt 2): 425–432.
PMCID: PMC1221082

Increase of mitochondria and mitochondrial DNA in response to oxidative stress in human cells.

Abstract

Mitochondrial respiratory function is impaired in the target tissues of patients with mitochondrial diseases and declines with age in various human tissues. It is generally accepted that respiratory-chain defects result in enhanced production of reactive oxygen species and free radicals in mitochondria. Recently, we have demonstrated that the copy number of mitochondrial DNA (mtDNA) is increased in the lung tissues of elderly human subjects. The mtDNA copy number was suggested to be increased by a feedback mechanism that compensates for defects in mitochondria harbouring mutated mtDNA and a defective respiratory system. However, the detailed mechanism remains unclear. In this study, we treated a human lung fibroblast cell line, MRC-5, with H(2)O(2) at concentrations of 90-360 microM. After the treatment for 24-72 h, we found that cells were arrested at G(0) and G(1) phases but that mitochondrial mass and mtDNA content were significantly increased in a concentration- and time-dependent manner. Moreover, the oxidative stress induced by buthionine sulphoximine was also found to cause an increase in mitochondrial mass of the treated cells. Increased uptake of a vital mitochondrial dye Rhodamine 123 and enhanced tetrazolium [MTT, 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] reduction revealed that the mitochondria increased by H(2)O(2) treatment were functional. In addition, the increase in the mitochondrial mass was also observed in cell-cycle-arrested cells induced by mimosine, lovastatin and genistein. Taken together, these findings suggest that the increase in mitochondrial mass and mtDNA content are the early molecular events of human cells in response to endogenous or exogenous oxidative stress through cell-cycle arrest.

Full Text

The Full Text of this article is available as a PDF (175K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Chance B, Sies H, Boveris A. Hydroperoxide metabolism in mammalian organs. Physiol Rev. 1979 Jul;59(3):527–605. [PubMed]
  • Papa S. Mitochondrial oxidative phosphorylation changes in the life span. Molecular aspects and physiopathological implications. Biochim Biophys Acta. 1996 Sep 12;1276(2):87–105. [PubMed]
  • Wei YH. Oxidative stress and mitochondrial DNA mutations in human aging. Proc Soc Exp Biol Med. 1998 Jan;217(1):53–63. [PubMed]
  • Lee HC, Lu CY, Fahn HJ, Wei YH. Aging- and smoking-associated alteration in the relative content of mitochondrial DNA in human lung. FEBS Lett. 1998 Dec 18;441(2):292–296. [PubMed]
  • Fahn HJ, Wang LS, Hsieh RH, Chang SC, Kao SH, Huang MH, Wei YH. Age-related 4,977 bp deletion in human lung mitochondrial DNA. Am J Respir Crit Care Med. 1996 Oct;154(4 Pt 1):1141–1145. [PubMed]
  • Fahn HJ, Wang LS, Kao SH, Chang SC, Huang MH, Wei YH. Smoking-associated mitochondrial DNA mutations and lipid peroxidation in human lung tissues. Am J Respir Cell Mol Biol. 1998 Dec;19(6):901–909. [PubMed]
  • Lee HC, Lim ML, Lu CY, Liu VW, Fahn HJ, Zhang C, Nagley P, Wei YH. Concurrent increase of oxidative DNA damage and lipid peroxidation together with mitochondrial DNA mutation in human lung tissues during aging--smoking enhances oxidative stress on the aged tissues. Arch Biochem Biophys. 1999 Feb 15;362(2):309–316. [PubMed]
  • Wei YH, Kao SH, Lee HC. Simultaneous increase of mitochondrial DNA deletions and lipid peroxidation in human aging. Ann N Y Acad Sci. 1996 Jun 15;786:24–43. [PubMed]
  • Lu CY, Lee HC, Fahn HJ, Wei YH. Oxidative damage elicited by imbalance of free radical scavenging enzymes is associated with large-scale mtDNA deletions in aging human skin. Mutat Res. 1999 Jan 25;423(1-2):11–21. [PubMed]
  • Gadaleta MN, Rainaldi G, Lezza AM, Milella F, Fracasso F, Cantatore P. Mitochondrial DNA copy number and mitochondrial DNA deletion in adult and senescent rats. Mutat Res. 1992 Sep;275(3-6):181–193. [PubMed]
  • Barrientos A, Casademont J, Cardellach F, Estivill X, Urbano-Marquez A, Nunes V. Reduced steady-state levels of mitochondrial RNA and increased mitochondrial DNA amount in human brain with aging. Brain Res Mol Brain Res. 1997 Dec 15;52(2):284–289. [PubMed]
  • Ames BN, Shigenaga MK, Hagen TM. Mitochondrial decay in aging. Biochim Biophys Acta. 1995 May 24;1271(1):165–170. [PubMed]
  • Sohal RS, Sohal BH. Hydrogen peroxide release by mitochondria increases during aging. Mech Ageing Dev. 1991 Feb;57(2):187–202. [PubMed]
  • Richter C, Gogvadze V, Laffranchi R, Schlapbach R, Schweizer M, Suter M, Walter P, Yaffee M. Oxidants in mitochondria: from physiology to diseases. Biochim Biophys Acta. 1995 May 24;1271(1):67–74. [PubMed]
  • Sohal RS, Sohal BH, Orr WC. Mitochondrial superoxide and hydrogen peroxide generation, protein oxidative damage, and longevity in different species of flies. Free Radic Biol Med. 1995 Oct;19(4):499–504. [PubMed]
  • Sen CK, Packer L. Antioxidant and redox regulation of gene transcription. FASEB J. 1996 May;10(7):709–720. [PubMed]
  • Lander HM. An essential role for free radicals and derived species in signal transduction. FASEB J. 1997 Feb;11(2):118–124. [PubMed]
  • Burdon RH. Superoxide and hydrogen peroxide in relation to mammalian cell proliferation. Free Radic Biol Med. 1995 Apr;18(4):775–794. [PubMed]
  • Poyton RO, McEwen JE. Crosstalk between nuclear and mitochondrial genomes. Annu Rev Biochem. 1996;65:563–607. [PubMed]
  • Scarpulla RC. Nuclear control of respiratory chain expression in mammalian cells. J Bioenerg Biomembr. 1997 Apr;29(2):109–119. [PubMed]
  • Lee HC, Wei YH. Mitochondrial role in life and death of the cell. J Biomed Sci. 2000 Jan-Feb;7(1):2–15. [PubMed]
  • Maftah A, Petit JM, Ratinaud MH, Julien R. 10-N nonyl-acridine orange: a fluorescent probe which stains mitochondria independently of their energetic state. Biochem Biophys Res Commun. 1989 Oct 16;164(1):185–190. [PubMed]
  • Petit JM, Maftah A, Ratinaud MH, Julien R. 10N-nonyl acridine orange interacts with cardiolipin and allows the quantification of this phospholipid in isolated mitochondria. Eur J Biochem. 1992 Oct 1;209(1):267–273. [PubMed]
  • Mancini M, Anderson BO, Caldwell E, Sedghinasab M, Paty PB, Hockenbery DM. Mitochondrial proliferation and paradoxical membrane depolarization during terminal differentiation and apoptosis in a human colon carcinoma cell line. J Cell Biol. 1997 Jul 28;138(2):449–469. [PMC free article] [PubMed]
  • Rothe G, Valet G. Flow cytometric analysis of respiratory burst activity in phagocytes with hydroethidine and 2',7'-dichlorofluorescin. J Leukoc Biol. 1990 May;47(5):440–448. [PubMed]
  • García-Ruiz C, Colell A, Morales A, Kaplowitz N, Fernández-Checa JC. Role of oxidative stress generated from the mitochondrial electron transport chain and mitochondrial glutathione status in loss of mitochondrial function and activation of transcription factor nuclear factor-kappa B: studies with isolated mitochondria and rat hepatocytes. Mol Pharmacol. 1995 Nov;48(5):825–834. [PubMed]
  • Pagliacci MC, Spinozzi F, Migliorati G, Fumi G, Smacchia M, Grignani F, Riccardi C, Nicoletti I. Genistein inhibits tumour cell growth in vitro but enhances mitochondrial reduction of tetrazolium salts: a further pitfall in the use of the MTT assay for evaluating cell growth and survival. Eur J Cancer. 1993;29A(11):1573–1577. [PubMed]
  • Robin ED, Wong R. Mitochondrial DNA molecules and virtual number of mitochondria per cell in mammalian cells. J Cell Physiol. 1988 Sep;136(3):507–513. [PubMed]
  • Renis M, Cantatore P, Loguercio Polosa P, Fracasso F, Gadaleta MN. Content of mitochondrial DNA and of three mitochondrial RNAs in developing and adult rat cerebellum. J Neurochem. 1989 Mar;52(3):750–754. [PubMed]
  • Shay JW, Pierce DJ, Werbin H. Mitochondrial DNA copy number is proportional to total cell DNA under a variety of growth conditions. J Biol Chem. 1990 Sep 5;265(25):14802–14807. [PubMed]
  • Williams RS, Salmons S, Newsholme EA, Kaufman RE, Mellor J. Regulation of nuclear and mitochondrial gene expression by contractile activity in skeletal muscle. J Biol Chem. 1986 Jan 5;261(1):376–380. [PubMed]
  • Bladier C, Wolvetang EJ, Hutchinson P, de Haan JB, Kola I. Response of a primary human fibroblast cell line to H2O2: senescence-like growth arrest or apoptosis? Cell Growth Differ. 1997 May;8(5):589–598. [PubMed]
  • Chen QM, Bartholomew JC, Campisi J, Acosta M, Reagan JD, Ames BN. Molecular analysis of H2O2-induced senescent-like growth arrest in normal human fibroblasts: p53 and Rb control G1 arrest but not cell replication. Biochem J. 1998 May 15;332(Pt 1):43–50. [PMC free article] [PubMed]
  • Suzuki H, Kumagai T, Goto A, Sugiura T. Increase in intracellular hydrogen peroxide and upregulation of a nuclear respiratory gene evoked by impairment of mitochondrial electron transfer in human cells. Biochem Biophys Res Commun. 1998 Aug 19;249(2):542–545. [PubMed]
  • Sazer S, Sherwood SW. Mitochondrial growth and DNA synthesis occur in the absence of nuclear DNA replication in fission yeast. J Cell Sci. 1990 Nov;97(Pt 3):509–516. [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...