Skip to main content
SpringerLink
Log in

In vitro cytotoxicity and induction of apoptosis by ruthenium(II) complexes in HL-60 and K-562

  • Original Research
  • Published:
Medicinal Chemistry Research Aims and scope Submit manuscript

Abstract

Mononuclear Ru complexes were evaluated for its cytotoxicity against two human leukemic cell lines such as acute myeloblastic leukemia (HL-60) and chronic myelogenic leukemia (K-562). Cytotoxicity was assayed by MTT assay. Fluorescent microscopic analysis, DNA fragmentation study by agarose gel electrophoresis, flow cytometric analysis, and caspase activity by spectrofluorimetry were carried out to evaluate the apoptosis-inducing property of Ru complexes. Ru complexes exhibited cytotoxicity against both the tested cell lines in a dose-dependent manner. Microscopic examination of ruthenium complex-treated cells showed characteristic morphological features of apoptosis, like cell shrinkage and formation of apoptotic bodies. Fluorescent staining with Hoechst 33342 revealed distinct chromatin condensation and nuclear fragmentation. DNA fragmentation pattern and flow cytometric analysis indicated that the cells treated with ruthenium complexes underwent apoptotic cell death. Thus from this investigation it may be concluded that Ru complexes act by the induction of apoptosis in tested human leukemic cell lines.

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
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

DNA:

Deoxyribonucleic acid

MTT:

3-(4,5-Dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide)

AFC:

7-Amino-4-trifluoromethylcoumarin

TEMED:

N,N,N′,N′-tetramethylethylene diamine

SDS:

Sodium dodecyl sulfate

CO2 :

Carbon dioxide

μl:

Microlitre

DMSO:

Dimethyl sulfoxide

PBS:

Phoshpate buffer solution

rpm:

Rotation per minute

HEPES:

4-(2-Hydroxy ethyl)-1-piperazine ethane sulfonic acid

NaCl:

Sodium chloride

MgCl2 :

Magnesium chloride

EDTA:

Ethylene diamine tetra acetic acid

RNase:

Ribonuclease

DNase:

Deoxyribonuclease

nm:

Nanometer

References

  • Arends MJ, Morris RJ, Wyllie AH (1990) Apoptosis: the role of endonuclease. Am J Pathol 136:593–608

    PubMed  CAS  Google Scholar 

  • Bergamo A, Gava B, Alessio E, Mestroni G, Serli B, Cocchieto M, Zoezet S, Sava G (2002) Ruthenium based NAMI-A type complexes with in vivo selective metastasis reduction and in vitro invasion inhibition unrelated to cell cytotoxicity. Int J Oncol 21:1331–1338

    PubMed  CAS  Google Scholar 

  • Cohen GM (1997) Caspases: the executioners of apoptosis. Biochem J 326:1–16

    PubMed  CAS  Google Scholar 

  • Compton MM (1992) A biochemical hallmark of apoptosis: internucleosomal degradation of the genome. Cancer Metastasis Rev 11:105–119

    Article  PubMed  CAS  Google Scholar 

  • Darzynkiewicz Z, Bruno S, Del Bino G, Gorczyca W, Hotz MA, Lassota P, Traganos F (1992) Features of apoptotic cells measured by flow cytometry. Cytometry 13:795–808

    Article  PubMed  CAS  Google Scholar 

  • Gupta M, Mazumder UK, Karki SS, Bhattacharya S, Suresh R, Sivakumar T (2004) Antineoplastic and antibacterial activity of some mononuclear Ru(II) complexes. J Enzym Inhib Med Chem 19:185–192

    Article  Google Scholar 

  • Hale AJ, Smith CA, Sutherland LC, Stoneman VEA, Longthrone VL, Culhane AC, Williams GT (1996) Apoptosis: molecular regulation of cell death. Eur J Biochem 236:1–26

    Article  PubMed  CAS  Google Scholar 

  • Hermann H, Lorenz HM, Voll R, Grunke M, Woith W, Kalden JR (1994) A rapid and simple method for the isolation of apoptotic DNA fragments. Nucleic Acids Res 22:5506–5507

    Article  Google Scholar 

  • Hotze AC, Caspers SE, De VD, Kooijman H, Spek AL, Flamigni A, Bacac M, Sava G, Haasnoot JG, Reedijk J (2004) Structure dependent in vitro cytotoxicity of the isomeric complexes [Ru(L)2Cl2] (L = o-tolylazopyridine and 4-methyl-2-phenyl-azopyridine) in comparison to [Ru(azpy)2Cl2]. J Biol Inorg Chem 9:354–364

    Article  PubMed  CAS  Google Scholar 

  • Keppler BK, Kreuser E, Berdel D, Wolfgang E, Slmuth P, Eckhard T (1992) Synergistic antitumor interactions between newly synthesized ruthenium complexes and cytokines in human colon carcinoma cell lines. Semin Oncol 19:73–81

    PubMed  Google Scholar 

  • Kerr JFR, Winterford CM, Harmon V (1994) Apoptosis, its significance in cancer and cancer therapy. Cancer 73:2013–2026

    Article  PubMed  CAS  Google Scholar 

  • Mazumder UK, Gupta M, Karki SS, Bhattacharya S, Suresh R, Sivakumar T (2004) Synthesis, anticancer and antibacterial activity of some novel mononuclear Ru(II) complexes. Chem Pharm Bull 52:178–185

    Article  PubMed  CAS  Google Scholar 

  • Mazumder UK, Gupta M, Karki SS, Bhattacharya S, Suresh R, Sivakumar T (2005) Synthesis and pharmacological activities of some mononuclear Ru(II) complexes. Bioorg Med Chem 13(20):5766–5773

    Article  PubMed  CAS  Google Scholar 

  • Nicholson DW, Thornberry NA (1997) Caspases: killer proteases. Trends Biochem Sci 22:299–306

    Article  PubMed  CAS  Google Scholar 

  • Nicoleti G, Oaggliacci MC, Grignanni F, Ricardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139:271–279

    Article  Google Scholar 

  • Roy M, Chakraborthy S, Siddiqui M, Bhattacharya RK (2002) Induction of apoptosis in tumor cells by natural phenolic compounds. Asia Pac J Cancer Prev 3:61–67

    Google Scholar 

  • Sava G, Cleriei K, Cocchietto M, Gagliardi R, Alessio E, Mestroni G, Perbellini A (1992) Effects of the Ru(III) complexes [mer-RuCl3(DMSO) and Na[trans-RuCl4(DMSO)Im] on solid mouse tumors. Anticancer Drugs 3:25–31

    Article  PubMed  CAS  Google Scholar 

  • Thornberry NA, Lazebnick Y (1998) Caspase: enemies within. Science 281:1312–1316

    Article  PubMed  CAS  Google Scholar 

  • Velders AH, Kooijman H, Spek AL, Haasnoot JG, De VD, Edijk J (2000) Strong differences in the in vitro cytotoxicity of three isomeric dichlorobis(2,2′-phenylazopyridine)ruthenium(II) complexes. Inorg Chem 39:2966–2967

    Article  PubMed  CAS  Google Scholar 

  • Wyllie AH, Kerr JFR, Currie AR (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68:251–307

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

This study was supported by the University Grant Commission of India.

Conflict of interest

Authors disclose that there is no conflict of interest.

Author information

Authors and Affiliations

  1. Division of Pharmaceutical Chemistry, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, West Bengal, 700 032, India

    S. Bhattacharya, R. Suresh, G. P. Senthilkumar & Upal K. Mazumder

  2. Division of Pharmacology, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, West Bengal, 700 032, India

    L. Manikandan & M. Gupta

  3. Department of Pharmaceutical Chemistry, KLE University’s College of Pharmacy, 2nd Block, Rajajinagar, Bangalore, Karnataka, 560 010, India

    Subhas S. Karki

  4. Department of Biotechnology, Dr. MGR Educational and Research Institute Maduravoyal, Chennai, Tamilnadu, 600 095, India

    R. Balaji

  5. Department of Pharmaceutical Chemistry, C.L. Baid Metha College of Pharmacy, Chennai, Tamilnadu, 600 097, India

    K. Murali

Authors
  1. S. Bhattacharya
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Subhas S. Karki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. Suresh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. L. Manikandan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. G. P. Senthilkumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M. Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Upal K. Mazumder
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. R. Balaji
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. K. Murali
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Upal K. Mazumder.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bhattacharya, S., Karki, S.S., Suresh, R. et al. In vitro cytotoxicity and induction of apoptosis by ruthenium(II) complexes in HL-60 and K-562. Med Chem Res 21, 1120–1126 (2012). https://doi.org/10.1007/s00044-011-9620-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00044-011-9620-6

Keywords

Search

Navigation