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Biological evaluation of a cytotoxic 2-substituted benzimidazole copper(II) complex: DNA damage, antiproliferation and apoptotic induction activity in human cervical cancer cells

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Abstract

Exploring novel chemotherapeutic agents is a great challenge in cancer medicine. To that end, 2-substituted benzimidazole copper(II) complex, [Cu(BMA)Cl2]·(CH3OH) (1) [BMA = N,N′-bis(benzimidazol-2-yl-methyl)amine], was synthesized and its cytotoxicity was characterized. The interaction between complex 1 and calf thymus DNA was detected by spectroscopy methods. The binding constant (K b = 1.24 × 104 M−1) and the apparent binding constant (K app = 6.67 × 106 M−1) of 1 indicated its moderate DNA affinity. Complex 1 induced single strand breaks of pUC19 plasmid DNA in the presence of H2O2 through an oxidative pathway. Cytotoxicity studies proved that complex 1 could inhibit the proliferation of human cervical carcinoma cell line HeLa in both time- and dose-dependent manners. The results of nuclei staining by Hoechst 33342 and alkaline single-cell gel electrophoresis proved that complex 1 caused cellular DNA damage in HeLa cells. Furthermore, treatment of HeLa cells with 1 resulted in S-phase arrest, loss of mitochondrial potential, and up-regulation of caspase-3 and -9 in HeLa cells, suggesting that complex 1 was capable of inducing apoptosis in cancer cells through the intrinsic mitochondrial pathway.

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Abbreviations

BMA:

N,N′-Bis(benzimidazol-2-yl-methyl)amine

CT-DNA:

Calf thymus DNA

DSBs:

Double strand breaks

EB:

Ethidium bromide

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

JC-1:

5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraatheylbenzamidazolo-carbocyanin iodide

LMPA:

Low melting point agarose

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PI:

Propidium iodide

PS:

Phosphatidylserine

ROS:

Reactive oxygen species

TAE:

Tris–acetate-EDTA

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Acknowledgments

We thank Tiffany K. West (Department of Cancer Biology, Wake Forest University Health Sciences, Winston-Salem, NC, USA) for language correction. This work was supported by National Natural Science Foundation of China (No. 21371135), the Key Program of Tianjin Municipal Natural Science Foundation (No. 13JCZDJC28200) and Innovation Fund for Graduate Students of Tianjin Medical University (No. 2010GSI05).

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Authors and Affiliations

  1. Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin, 300070, People’s Republic of China

    Xin Qiao, Zhong-Ying Ma, Jia Shao, Wei-Guo Bao & Jing-Yuan Xu

  2. School of Basic Medicine, Tianjin Medical University, Tianjin, 300070, People’s Republic of China

    Zhao-Yan Qiang & Jian-Shi Lou

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  1. Xin Qiao
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  2. Zhong-Ying Ma
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Correspondence to Jing-Yuan Xu or Jian-Shi Lou.

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Qiao, X., Ma, ZY., Shao, J. et al. Biological evaluation of a cytotoxic 2-substituted benzimidazole copper(II) complex: DNA damage, antiproliferation and apoptotic induction activity in human cervical cancer cells. Biometals 27, 155–172 (2014). https://doi.org/10.1007/s10534-013-9696-1

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