Skip to main content

Advertisement

SpringerLink
Log in

Tetrazolium violet induces G0/G1 arrest and apoptosis in brain tumor cells

  • Laboratory Investigation-human/animal tissue
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Summary

Tetrazolium violet (TV), a potent anticancer agent, has been shown to induce cell growth-inhibition in tumor cells. However, the related mechanism has not been revealed yet. In this report we assessed the influence of TV on cell growth and cell cycle in brain tumor cells. Treatment of C6 tumor cells with TV (5–15 μM for 24–72 h) resulted in a growth inhibition in a dose and time-dependent manner and G0/G1 phase arrest, determined by flow cytometry analysis. These effects were accompanied by apoptosis other than necrosis, evidenced by nuclear condensation, terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay and trypan blue exclusion assay plus lactate dehydrogenase (LDH) release assay. Treatment of cells with TV at 15 μM for 24 h resulted in an increase in the activity of caspase-3, evidenced by colorimetric assay, and a dramatic up-regulation of p53, accompanied with a significant increase of Bax/Bcl-2 ratio, as evidenced by immunofluorescence assay. These results suggest that TV induces growth inhibition of C6 cells through p53-midiated apoptotic pathway and G0/G1 checkpoint mechanism. Although detailed mechanisms remain to be explored, selective blockage of tumor cells in G0/G1 phase accompanied by p53-associated apoptosis makes tetrazolium violet a promising anticancer agent, meriting further investigations.

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

Similar content being viewed by others

Abbreviations

TV:

tetrazolium violet

C6:

cell from rat glial tumor

9L:

rat gliosarcoma cell line

CNS-1:

inbred Lewis rat glioma cell line

TUNEL:

TdT-mediated nick end labeling

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide

PBS:

phosphate-buffered saline

DMEM:

Dulbecco’s modified Eagle’s medium

HEPES:

4-(2-hydroxyerhyl)piperazine-1-erhanesulfonicacid

FBS:

fetal bovine serum

LDH:

lactate dehydrogenase

FACS:

fluorescence-activated cell sorter

FITC:

fluorescein isothiocyanate

TRITC:

tetramethylrhodamine isothiocyanante

CDK:

cyclin-dependent kinase

Ig:

immunoglobulin.

References

  1. Thompson CB, 1995 Apoptosis in the pathogenesis and treatment of disease Science 267: 1456–1462

    Article  PubMed  CAS  Google Scholar 

  2. Steller H, 1995 Mechanisms and genes of cellular suicideScience 267:1445–1449

    Article  PubMed  CAS  Google Scholar 

  3. Fisher DE, 1994 Apoptosis in cancer therapy: crossing the thresholdCell 78: 539–542

    Article  PubMed  CAS  Google Scholar 

  4. Bouton LA, Ramirez CD, Bailey DP, Yeatman CF, Yue J, Wright HV, Domen J, Rosato RR, Grant S, Fischer-Stenger K, Ryan JJ, 2004 Costimulation with interleukin-4 and interleukin-10 induces mast cell apoptosis and cell-cycle arrest: the role of p53 and the mitochondrionExp Hematol 32(12):1137–45

    Article  PubMed  CAS  Google Scholar 

  5. Kong Q, Zhang N, Zhao YF: Tetrazolium violet sensitizes tumor cells to chemotherapeutic agents. Proceedings of the American Association for Cancer Research A556, 2004

  6. Price P, McMillan TJ, 1990 Use of the tetrazolium assay in measuring the response of human Tumor cells to ionizing radiationCancer Res 50: 1392–1396

    PubMed  CAS  Google Scholar 

  7. Sugiyama K, Akiyama T, Shimizu M, Tamaoki T, Carol Courage, Gescher A, Akinaga S, 1999 Decrease in susceptibility toward induction of apoptosis and alteration in G1 checkpoint function as determinants of resistance of human lung cancer cells against the antisignaling drug UCN-01 (7-Hydroxystaurosporine) Cancer Res 59: 4406–4412

    PubMed  CAS  Google Scholar 

  8. Fueyo J, Gomez-Manzano C, Yung WK, Liu TJ, Alemany R, McDonnell TJ, Shi X, Rao JS, Levin VA, Kyritsis AP, 1998 Overexpression of E2F-1 in glioma triggers apoptosis and suppresses tumor growth in vitro and in vivoNat Med 4:685–90

    Article  PubMed  CAS  Google Scholar 

  9. Ruan MS, Okcu F, Pong R-C, Andreeff M, Levin V, Hsieh J-T, Zhang W, 1999 Attenuation of WAF1/Cip1 expression by an antisense adenovirus expression vector sensitizes glioblastoma cells to apoptosis induced by chemotherapeutic agents 1,3-Bis(2- chloroethyl)-1-nitrosourea and CisplatinClinl Cancer Res 5: 197–202

    CAS  Google Scholar 

  10. Gobert C, Skladanowski A, Larsen AK, 1999 The interaction between p53 and DNA topoisomerase I is regulated differently in cells with wild-type and mutant p53. PNAS 96(18): 10355– 10360

    Article  PubMed  CAS  Google Scholar 

  11. Lau WK, Yeung CW, Lui PW, Cheung LH, Poon NT, Yung KKL, 2002 Different trends in modulation of NMDAR1 and NMDAR2B gene expression in cultured cortical and hippocampal neurons after lead exposureBrain Res 932: 10–20

    Article  PubMed  CAS  Google Scholar 

  12. Wyllie AH, Kerr JF, Currie AR, 1980 Cell death: the significance of apoptosis. Int Rev Cytol 68: 251–306

    Article  PubMed  CAS  Google Scholar 

  13. Bezabeh T, Mowat MRA, Jarolim L, Greenberg AH, Smith ICP, 2001 Detection of drug-induced apoptosis and necrosis in human cervical carcinoma cells using 1H NMR spectroscopyCell Death Differ 8: 219–224

    Article  PubMed  CAS  Google Scholar 

  14. Sun J, Kong L D, Kong Q: Centrosome crystallization by tetrazolium salts for cancer therapy. American Association for the Advancement of Science, Meeting Program A96, 2002

  15. O’Connor PM, 1997 Mammalian G1 and G2 phase checkpointsCancer Surv 29:151–182

    PubMed  CAS  Google Scholar 

  16. Hartwell LH, Kastan MB, 1994 Cell cycle control and cancerScience 266: 1821–1828

    Article  PubMed  CAS  Google Scholar 

  17. Fulda S, Debatin KM, 2004 Sensitization for anticancer drug-induced apoptosis by the chemopreventive agent resveratrolOncogene 23(40): 6702–6711

    Article  PubMed  CAS  Google Scholar 

  18. Yoon HS, Chen X, Yang VW, 2003 Kru ppel-like Factor 4 Mediates p53-dependent G1/S Cell Cycle Arrest in Response to DNA DamageJ Biol chem 278: 2101–2105

    Article  PubMed  CAS  Google Scholar 

  19. Mercer J, Figg N, Stoneman V, Braganza D, Bennett M, 2005 Endogenous p53 protects vascular smooth muscle cells from apoptosis and reduces atherosclerosis in ApoE knockout miceCirc Res 96(6):667–674

    Article  PubMed  CAS  Google Scholar 

  20. Bissonnette N, Wasylyk B, Hunting DJ, 1997 The apoptotic and transcriptional transactivation activities of p53 can be dissociatedBiochem Cell Biol 75: 351–358

    Article  PubMed  CAS  Google Scholar 

  21. Wlodarski BP, Wasik M, Ratajczak MZ, Sevignani C, Hoser G, Kawiak J, Gewirtz AM, Calabretta B, Skorski T, 1998 Role of p53 in hematopoietic recovery after cytotoxic treatmentBlood 91: 2998–3006

    PubMed  CAS  Google Scholar 

  22. Bates S, Vousden KH, 1999 Mechanism of p53-mediated apoptosisCell Mol Life Sci 55: 28–37

    Article  PubMed  CAS  Google Scholar 

  23. Sabbatini P, Han J, Chiou S, Nicholson DW, White E, 1997 Interleukin 1beta converting enzyme-like protease are essential for p53-mediated transcriptionally dependent apoptosisCell Growth Differ 8: 643–653

    PubMed  CAS  Google Scholar 

  24. Ding HF, McGill G, Rowan S, Schmalz C, Shimamura A, Fisher DE, 1998 Oncogene-dependent regulation of caspase activation by p53 protein in a cell-free systemJ Biol Chem 273: 28378– 28383

    Article  PubMed  CAS  Google Scholar 

  25. Gong XF, Wang MW, Tashiro S, Onodera S, Ikejima T, 2005 Pseudolaric acid B induces apoptosis through p53 and Bax/Bcl-2 pathways in human melanoma A375-S2 cellsArch Pharm Res 28(1): 68–72

    Article  PubMed  CAS  Google Scholar 

  26. Kong Q, 2003 The centrosome-centered Cell-brain in apoptosisMed Hypotheses 61(1): 126–132

    Article  PubMed  CAS  Google Scholar 

  27. Kong Q, Zhang N, Zhao YF, 2004 The roles of centrosome in cancer developmentDrug News Perspec 17:195–200

    Article  CAS  Google Scholar 

  28. Hinchcliffe EH, Miller FJ, Cham M, Khodjakov A, Sluder G, 2001 Requirement of a Centrosomal Activity for Cell Cycle Progression Through G1 into S PhaseScience 291:1547–1550

    Article  PubMed  CAS  Google Scholar 

  29. Khodjakov A, Rieder CL, 2001 Centrosomes enhance the fidelity of cytokinesis in vertebrates and are required for cell cycle progressionJ Cell Biol 153(1): 237–42

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Life Science, Shandong University, Jinan, 250100, China

    Yunfeng Zhao & Qingzhong Kong

  2. School of Life Science, Qufu Normal University, Qufu, Qufu 273165, China

    Yunfeng Zhao

  3. Department of Medicines, Jinan Central Hospital, Jinan, Shandong Province, China

    Nan Zhang

  4. LanJin Biotech Company, Jinan, 250100, China

    Qingzhong Kong

Authors
  1. Yunfeng Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qingzhong Kong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Qingzhong Kong.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhao, Y., Zhang, N. & Kong, Q. Tetrazolium violet induces G0/G1 arrest and apoptosis in brain tumor cells. J Neurooncol 77, 109–115 (2006). https://doi.org/10.1007/s11060-005-9012-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11060-005-9012-1

Keywords

Search

Navigation