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DNA binding, cleavage and cytotoxicity studies of three mononuclear Cu(II) chloro-complexes containing N–S donor Schiff base ligands

  • Sidhali U. Parsekar
  • Joseph Fernandes
  • Arnab Banerjee
  • Om Prakash Chouhan
  • Sumit Biswas
  • Manohar Singh
  • Durga P. Mishra
  • Manjuri Kumar
Original Paper
  • 38 Downloads

Abstract

We report the biological activity of three Cu(II) complexes [Cu(pabt)Cl] (1), [Cu(pma)Cl] (2), and [Cu(pdta)Cl]Cl (3) (pabt = N-(2-mercaptophenyl)-2′-pyridylmethylenimine, pma = N-(2-pyridylmethyl)-2-mercaptoaniline, pdta = 2,2′-di(pyridyl-2-methyleneimine)diphenyl disulfide). 13 display four-line EPR multiplet in solution at RT suggesting that these are mononuclear. DNA-binding studies using spectrophotometric titration of these complexes with calf thymus DNA showed binding through intercalation mode which was found to be consistent with the observation of increased viscosity of DNA and quenching of fluorescence of ethidium bromide bound DNA in the presence of these complexes. All three complexes were found to be efficient in bringing about oxidative and hydrolytic cleavage of DNA. The proposed mechanism of hydrolytic DNA cleavage has been discussed. MTT assay showed remarkable cytotoxicity on cervical cancer HeLa cell line and the IC50 values were 1.27, 4.13, and 3.92 μM for 1, 2 and 3, respectively, as compared to the IC50 value (13 μM) reported for cisplatin in HeLa cells. AO/PI and Annexin-V/PI assay suggest the induction of cell death primarily via apoptotic pathway. Nuclear staining using DAPI was used to assess changes in nuclear morphology during apoptotic cell death. The role of reactive oxygen species (ROS) for induction of apoptotic cell death was studied using H2DCF-DA assay and the result suggests that the generation of ROS by the complexes may be a possible cause for their antiproliferative activity. TUNEL assay showed DNA fragmentation in apoptotic cells. Cell cycle analysis using flow cytometry showed significant increase in the G2/M phase in HeLa cells by the compounds 13.

Graphical abstract

Mononuclear Cu(II) complexes display remarkable cytotoxicity against cervical cancer HeLa cell line. The generation of ROS by the complexes may be a cause of their antiproliferative activity. Fluorescent images from DAPI staining assay revealed that the cells undergoing apoptosis displayed typical features like cell shrinkage, membrane blebbing, chromatin condensation and nuclear fragmentation. TUNEL assay showed DNA fragmentation in apoptotic cells.

Keywords

Cu(II) complexes DNA binding and cleavage MTT assay TUNEL assay Cytotoxicity Anticancer activity 

Notes

Acknowledgements

We thank SAIF, IIT-Madras for EPR facilities and SAIF, IIT-Bombay for recording ESI MS for the compounds 1 and 3 with Varian Inc., USA Liquid Chromatograph Mass Spectrometer Model 410 Prostar Binary LC with 500 MS IT PDA Detectors. M.K. thanks Ms. Neha Saran and Prof. P. Bhavana, Department of Chemistry, BITS-Pilani K. K. Birla Goa Campus for help with the electrochemical studies.

Supplementary material

775_2018_1620_MOESM1_ESM.pdf (4.7 mb)
Supplementary material Fig. S1 ESI-MS spectrum of 1, 2 and 3 in CH3CN. Fig. S2 X-band RT solution EPR spectra of 1-3. Fig. S3 Cyclic voltammograms for 1-3 in CH3CN. Fig. S4 Effect of increasing amount of 1-3 on the relative viscosity of CT-DNA (100 μM) in 10 mM Tris.HCl buffer (pH 7.4) at room temperature. Fig. S5 ROS detection in HeLa cells treated with H2O2 (final concentration of 180 µM) as a positive control. Fig. S6 Detection of apoptotic DNA fragmentation by TUNEL assay using Alexa Fluor® picolyl azide dye (PDF 4793 kb)

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Copyright information

© SBIC 2018

Authors and Affiliations

  • Sidhali U. Parsekar
    • 1
  • Joseph Fernandes
    • 2
  • Arnab Banerjee
    • 2
  • Om Prakash Chouhan
    • 2
  • Sumit Biswas
    • 2
  • Manohar Singh
    • 3
  • Durga P. Mishra
    • 3
  • Manjuri Kumar
    • 1
  1. 1.Department of Chemical EngineeringBirla Institute of Technology and Science-PilaniGoaIndia
  2. 2.Department of Biological SciencesBirla Institute of Technology and Science-PilaniGoaIndia
  3. 3.Cell Death Research Laboratory, Endocrinology DivisionCSIR-Central Drug Research InstituteLucknowIndia

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