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A natural flavonoid, apigenin isolated from Clerodendrum viscosum leaves, induces G2/M phase cell cycle arrest and apoptosis in MCF-7 cells through the regulation of p53 and caspase-cascade pathway

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Abstract

Background

With 9.6 million deaths in 2018, cancer remains the second leading cause of death worldwide. Breast cancer is the most deadly type of cancer among females, with 55.2% of crude incidence rate and 16.6% of crude mortality rate.

Purpose

The present study was aimed to investigate the anti-breast cancer potential of natural dietary flavonoid, apigenin isolated from Clerodendrum viscosum leaves.

Methods

Apigenin was evaluated for in-depth anticancer activity in MCF-7 cells using cell viability assay, cell cycle analysis, Annexin-V-FLUOS staining, ROS induction, morphological analysis, and western blot analysis.

Results

Apigenin showed selective cytotoxicity on MCF-7 cells with an IC50-56.72 ± 2.35 µM, while negligible cytotoxicity was observed on WI-38 cells. Further, the flow cytometer-based analysis showed that apigenin halted MCF-7 cells in the G2/M phase arrest followed by dose-dependent apoptosis. Moreover, the FACS and confocal microscopy results confirmed the elevation of intracellular ROS and nuclear fragmentation in apigenin-treated MCF-7 cells. Western blots showed up-regulation of cell cycle regulatory proteins, increased p53 expression, Bax/Bcl-2 ratio, activation of caspases, and cleavage of PARP. Finally, apigenin treatment in the presence of Pifithrin-µ showed decreased apoptotic population and it was further confirmed through western blotting study. The results revealed the vital role of p53 in apigenin-induced apoptosis in MCF-7 cells.

Conclusions

In the present findings, treatment of apigenin-induced intracellular ROS in MCF-7 cells followed by induction of G2/M phase cell cycle arrest and further apoptosis through the regulation of p53 and caspase-cascade signaling pathway.

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Acknowledgements

AKS and DC are grateful to the Council of Scientific and Industrial Research (CSIR) Govt. of India and Tapasree Basu is grateful to the University Grants Commission (UGC), Govt. of India for providing the fellowships. The authors are thankful to Dr. Nikhil Baban Ghate for his constant guidance during the experiments and Mr. Ranjit Kumar Das for his technical assistance.

Funding

This research did not receive any specific grant from funding agencies.

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Authors and Affiliations

  1. Division of Molecular Medicine, Bose Institute, P-1/12 CIT Scheme - VIIM, Kolkata, West Bengal, 700054, India

    A. K. Shendge, D. Chaudhuri, T. Basu & N. Mandal

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  1. A. K. Shendge
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  2. D. Chaudhuri
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  3. T. Basu
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Correspondence to N. Mandal.

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AKS, DC, TB, and NM declared no conflict of interest.

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Shendge, A.K., Chaudhuri, D., Basu, T. et al. A natural flavonoid, apigenin isolated from Clerodendrum viscosum leaves, induces G2/M phase cell cycle arrest and apoptosis in MCF-7 cells through the regulation of p53 and caspase-cascade pathway. Clin Transl Oncol 23, 718–730 (2021). https://doi.org/10.1007/s12094-020-02461-0

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