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Centchroman induces redox-dependent apoptosis and cell-cycle arrest in human endometrial cancer cells

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Abstract

Centchroman (CC) or Ormeloxifene has been shown to induce apoptosis and cell cycle arrest in various types of cancer cells. This has, however, not been addressed for endometrial cancer cells where its (CC) mechanism of action remains unclear. This study focuses on the basis of antineoplasticity of CC by blocking the targets involved in the cell cycle, survival and apoptosis in endometrial cancer cells. Ishikawa Human Endometrial Cancer Cells were cultured under estrogen deprived medium, exposed to CC and analyzed for proliferation and apoptosis. Additionally, we also analyzed oxidative stress induced by CC. Cell viability studies confirmed the IC50 of CC in Ishikawa cells to be 20 µM after 48 h treatment. CC arrests the cells in G0/G1 phase through cyclin D1 and cyclin E mediated pathways. Phosphatidylserine externalization, nuclear morphology changes, DNA fragmentation, PARP cleavage, and alteration of Bcl-2 family protein expression clearly suggest ongoing apoptosis in the CC treated cells. Activation of caspase 3 & 9, up-regulation of AIF and inhibition of apoptosis by z-VAD-fmk clearly explains the participation of the intrinsic pathway of programmed cell death. Further, the increase of ROS, loss of MMP, inhibition of antioxidant (MnSOD, Cu/Zn-SOD and GST) and inhibition of apoptosis with L-NAC suggests CC induced oxidative stress leading to apoptosis via mitochondria mediated pathway. Therefore, CC could be a potential therapeutic agent for the treatment of Endometrial Cancer adjunct to its utility as a contraceptive and an anti-breast cancer agent.

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Acknowledgements

The authors are thankful to the Director, CSIR-CDRI for permitting to carry out the work. Mr. A. L. Vishwakarma, SAIF is thanked for the experiments with flow cytometry. Hari Shyam and Shweta Kaushik gratefully acknowledge the fellowship received from ICMR and UGC New Delhi, respectively. This is the CSIR-CDRI communication no. 9428.

Author Contributions

Conception and design: Hari Shyam, Neetu Singh and Anil K. Balapure. Development of methodology: Hari Shyam, Neetu Singh and Anil K.Balapure. Acquisition of data: Hari Shyam, Shweta Kaushik and Ramesh Sharma. Shyam, Neetu Singh, Ramesh Sharma and Anil K. Balapure. Writing, review and/or revision of the manuscript: Hari Shyam, Neetu Singh and Anil K. Balapure. Study supervision: Anil K. Balapure.

Funding

This study was supported by Project code BSC0001, CSIR-Central Drug Research Institute, Lucknow, India and Indian Council of Medical Research, New Delhi, India.

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Authors and Affiliations

  1. Division of Biochemistry, CSIR-Central Drug Research Institute, Council of Scientific & Industrial Research, 226031, Lucknow, India

    Hari Shyam, Shweta Kaushik, Ramesh Sharma & Anil K. Balapure

  2. Department of Molecular Biology, Centre for Advance Research, King George’s Medical University, 226 003, Lucknow, India

    Hari Shyam & Neetu Singh

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  1. Hari Shyam
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  2. Neetu Singh
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  3. Shweta Kaushik
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  4. Ramesh Sharma
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  5. Anil K. Balapure
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Correspondence to Anil K. Balapure.

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The authors declare no potential conflict of Interest.

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Hari Shyam is the recipient of a research fellowship from the Indian Council of Medical Research, and this work constitutes a part of his PhD thesis.

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Shyam, H., Singh, N., Kaushik, S. et al. Centchroman induces redox-dependent apoptosis and cell-cycle arrest in human endometrial cancer cells. Apoptosis 22, 570–584 (2017). https://doi.org/10.1007/s10495-017-1346-6

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  • DOI: https://doi.org/10.1007/s10495-017-1346-6

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