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Gedunin isolated from the mangrove plant Xylocarpus granatum exerts its anti-proliferative activity in ovarian cancer cells through G2/M-phase arrest and oxidative stress-mediated intrinsic apoptosis

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Abstract

Gedunin is a natural tetranorterpenoid secondary metabolite found in plants of the Meliaceae family, which has been reported for its antiparasitic, antifungal and anticancer activities. Here, we describe the molecular mechanisms underlying the in vitro anti proliferative activity of gedunin (isolated from the mangrove plant Xylocarpus granatum) in human ovarian cancer cells. We observed that gedunin triggered severe ROS generation leading to DNA damage and cell cycle arrest in G2/M phase thus inhibiting cell proliferation. ROS upregulation also led to mitochondrial stress and membrane depolarization, which eventually resulted in mitochondria-mediated apoptosis following cytochrome C release, caspase 9, 3 activation, and PARP cleavage. Transmission electron microscopy of gedunin treated cells revealed sub-cellular features typical of apoptosis. Moreover, an upregulation in stress kinases like phospho-ERK 1/2, phospho-p38 and phospho-JNK was also observed in gedunin treated cells. Free radical scavenger N-Acetyl-L-Cysteine (NAC) reversed all these effects resulting in increased cell survival, abrogation of cell cycle arrest, rescue of mitochondrial membrane potential and suppression of apoptotic markers. Interestingly, gedunin is also an inhibitor of the evolutionarily conserved molecular chaperone Heat Shock Protein 90 (hsp90) responsible for maintaining cellular homeostasis. Targeting this chaperone could be an attractive strategy for developing cancer therapeutics since many oncogenic proteins are also client proteins of hsp90. Collectively, our findings provide insights into the molecular mechanism of action of gedunin, which may aid drug development efforts against ovarian cancer.

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Acknowledgements

Technical assistance from Ms. Garima Pant and Mrs. Madhuli Srivastava for EM specimen preparation at EM Unit SAIF-R CDRI; and Mr. AL Vishwakarma and Mrs. Madhu Chaturvedi for flow-cytometer analysis, SAIF-R CDRI is acknowledged. This is CDRI communication number 10065.

Funding

Funding support from CSIR network project BSC0120 and institutional project OLP0101 to KM, fellowship DBT-SRF to RS, CSIR-SRF to AB, CSIR-SRF to PY and ICMR-SRF to MH is duly acknowledged.

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Authors and Affiliations

  1. Electron Microscopy Unit, Sophisticated Analytical Instrument Facility and Research, CSIR – Central Drug Research Institute, Sector-10, Jankipuram Extension, Lucknow, Uttar Pradesh, 226 031, India

    Rohit Sahai, Arindam Bhattacharjee & Kalyan Mitra

  2. Medicinal and Process Chemistry Division, CSIR – Central Drug Research Institute, Sector-10, Jankipuram Extension, Lucknow, Uttar Pradesh, 226 031, India

    Vishwa Nath Shukla, Pragya Yadav & T. Narender

  3. Division of Biochemistry, CSIR – Central Drug Research Institute, Sector-10, Jankipuram Extension, Lucknow, Uttar Pradesh, 226 031, India

    Mohammad Hasanain & Jayanta Sarkar

  4. Academy of Scientific and Innovative Research, Ghaziabad, Uttar Pradesh, 201 002, India

    Pragya Yadav, Mohammad Hasanain, Jayanta Sarkar, T. Narender & Kalyan Mitra

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  1. Rohit Sahai
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  2. Arindam Bhattacharjee
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Contributions

RS performed most of the experiments, prepared figures and wrote the first draft of the manuscript; AB performed some experiments and edited manuscript; VNS, PY, TN isolated gedunin; JS provided critical inputs and shared resources; MH performed cell viability assay supervised by JS; TN and KM conceived the study; KM supervised the study, interpreted data, arranged funding, edited and finalized the manuscript.

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Correspondence to Kalyan Mitra.

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Sahai, R., Bhattacharjee, A., Shukla, V.N. et al. Gedunin isolated from the mangrove plant Xylocarpus granatum exerts its anti-proliferative activity in ovarian cancer cells through G2/M-phase arrest and oxidative stress-mediated intrinsic apoptosis. Apoptosis 25, 481–499 (2020). https://doi.org/10.1007/s10495-020-01605-5

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