Carvacrol Promotes Cell Cycle Arrest and Apoptosis through PI3K/AKT Signaling Pathway in MCF-7 Breast Cancer Cells

Abstract

Objective

To examine the role of carvacrol in modulating PI3K/AKT signaling involved in human breast cancer pathogenesis using in vitro experimental model MCF-7 cells.

Methods

MTT and lactate dehydrogenase assays were performed with cells treated with different doses of carvacrol (0-250 p mol/L) at different time points (24 and 48 h). The nuclear morphology was assessed in MCF-7 cells with propidium iodide (PI) and acridine orange/ethidium bromide (AO/EB) staining and analyzed by fluorescence microscopy. Events like cell cycle arrest, apoptosis was observed by flow cytometric analysis and expressions of p-Rb, cyclin D1, cyclin-dependent kinase 4 (CDK4), CDK6, Bax, Bcl-2, PI3K/p-AKT was analyzed by immunoblot.

Results

Carvacrol significantly reduced cell viability with the half maximal inhibitory concentration value of 200 µmol/L at 24 and 48 h (P<0.05). importantly, there was a significant increase in the accumulation of the G0/G1 phase upon treatment with carvacrol in MCF-7 cells (P<0.05 or P<0.01). A remarkable decrease in protein expressions of p-Rb, cyclin D1, CDK4 and CDK6 denotes cell cycle arrest (P<0.05 or P<0.01). In addition, carvacrol treatment significantly inhibited PI3K/p-AKT protein expressions leading to induction of apoptosis mediated by decreased Bcl2 and increased Bax protein expressions. Further, Annexin V/PI staining by FACS analysis, dual staining by AO/EB and PI staining studies suggests induction of apoptosis by carvacrol through PI3K/Akt signaling pathway in MCF-7 cells.

Conclusion

Carvacrol significantly inhibited the breast cancer MCF-7 cell proliferation and induced apoptosis via suppressing PI3/AKT signaling pathway.

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Acknowledgment

First author Mr. M. Ashok is grateful to the Indian Council of Medical Research (ICMR) New Delhi, India for the support in the form of Senior Research Fellowship (SRF).

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Authors

Contributions

Mari A collected the samples, carried out all the experiential work, has done all data collection, interpretation and drafted the manuscript; Thiruvengadam D contributed to study design, results interpretation and manuscript draft; Nagabhishek SN contributed to cell culture assays, interpretation and manuscript draft; Mani G and Balaraman G helped carry out studies on flow cytometry; Subramanian N, Mirza FB and Sundaram J helped carry out Western blot studies. All the authors read and approved the final manuscript.

Corresponding author

Correspondence to Devaki Thiruvengadam.

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Conflict of Interest

The authors declare that they have no conflict of interest.

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Mari, A., Mani, G., Nagabhishek, S.N. et al. Carvacrol Promotes Cell Cycle Arrest and Apoptosis through PI3K/AKT Signaling Pathway in MCF-7 Breast Cancer Cells. Chin. J. Integr. Med. (2020). https://doi.org/10.1007/s11655-020-3193-5

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Keywords

  • carvacrol
  • breast cancer
  • proliferation
  • apoptosis
  • PI3K/AKT signaling pathway