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Novel combination of salinomycin and resveratrol synergistically enhances the anti-proliferative and pro-apoptotic effects on human breast cancer cells

Abstract

Resveratrol (RES) is a natural polyphenol having anti-proliferative activity against breast cancer cells. RES in combination with other chemo modulatory agents, minimizes toxicity and increases efficacy of the treatment. Salinomycin (SAL), a monocarboxylic polyether ionophore is known for selectively targeting breast cancer stem cells. Purpose of the present study was to investigate whether RES in combination with SAL exerts synergistic anti-proliferative activity on breast cancer cells. We further evaluated the molecular mechanism behind SAL and RES mediated cell death. Cytotoxicity assay was performed to determine 50% inhibitory concentration (IC50) of SAL and RES in different human breast cancer cells (HBCCs). Drug synergism and combination index (CI) were calculated using CompuSyn software and effects of synergistic combinations (CI < 1) involving lower doses of SAL and RES were selected for further studies. This combination significantly induced apoptosis in HBCCs without affecting non tumorigenic human breast epithelial cells MCF-10A. Co-treatment enhanced apoptosis in MCF-7 cells via reactive oxygen species (ROS) mediated mitochondrial dysfunction. Oxidative stress disrupt redox homeostasis which altered antioxidant enzymes viz. CuZn Superoxide dismutase (SOD), MnSOD and catalase. Additionally, combination altered nuclear morphology, enhanced PARP cleavage and led to caspase activation. SAL and RES also synergistically modulated MAPK pathway. Study suggests that SAL and RES offer a novel combination approach for the treatment of breast cancer.

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Acknowledgements

This work was supported by grants from Council of Scientific and Industrial Research, networking project BSC0103. J.D. gratefully acknowledge the fellowship received from University Grant Commission, New Delhi, A.K.V. and D.T. acknowledge the fellowship received from Council of Scientific and Industrial Research, S.S. acknowledge the fellowship received from Indian Council of Medical Research. Authors want to acknowledge Mr. A.L. Vishwakarma for support in flow cytometric studies. We are greatly thankful to Dr. Sharad Sharma for his valuable help in English editing of this manuscript. CSIR-CDRI communication number for this manuscript is 9514.

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JD and SKR designed study, analysed data and prepared draft of manuscript. JD, DT, AY performed experimental work. SS and AKV helped in flow cytometer and data interpretation. All authors reviewed the manuscript.

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Correspondence to Srikanta Kumar Rath.

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The authors declare no competing financial interests.

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Dewangan, J., Tandon, D., Srivastava, S. et al. Novel combination of salinomycin and resveratrol synergistically enhances the anti-proliferative and pro-apoptotic effects on human breast cancer cells. Apoptosis 22, 1246–1259 (2017). https://doi.org/10.1007/s10495-017-1394-y

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  • DOI: https://doi.org/10.1007/s10495-017-1394-y

Keywords

  • Breast cancer
  • Salinomycin
  • Resveratrol
  • Apoptosis
  • Synergism
  • MAPK