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



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.


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.


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.


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

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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).

Author information




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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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).

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