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Cell cycle arrest-mediated cell death by morin in MDA-MB-231 triple-negative breast cancer cells

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Abstract

Background

Morin, a flavonoid extracted from Moraceace family and exhibits several pharmacological activities including anti-cancer activity. Although the anticancer activity of morin in breast cancer was estimated in some investigations, the pharmaceutical mechanism has not been fully elucidated. Therefore, we investigated to unveil the detail signaling pathway in morin-treated in MDA-MB-231 triple-negative breast cancer cells.

Methods

The cytotoxicity of morin in MDA-MB-231 cells was confirmed by sulforhodamine B (SRB) assay and colony formation assay. Flow cytometry was performed to examine the cell cycle and cell death patterns and the protein expression and phosphorylation were detected by western blotting.

Results

Our results showed that morin inhibited MDA-MB-231 cells proliferation in time and concentration-dependent manner. Morphological changes were observed when treated with various concentration of morin in MDA-MB-231 cells. In regard to protein expression, morin induced the phosphorylation of ERK and p-H2A.X and decreased the level of DNA repair markers, RAD51 and survivin. In addition, flow cytometry showed S and G2/M arrest by morin that was associated with the decrease in the protein expression of cyclin A2 and cyclin B1 and upregulation of p21. Interestingly, annexin V/PI staining result clearly showed that morin induced cell death without apoptosis. Furthermore, attenuated FoxM1 by morin was co-related with cell cycle regulators including p21, cyclin A2 and cyclin B1.

Conclusion

Taken together, our study indicates that morin-induced cell death of MDA-MB-231 is caused by sustained cell cycle arrest via the induction of p21 expression by activation of ERK and repression of FOXM1 signaling pathways.

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Abbreviations

AIF:

Apoptosis-inducing factor

BAX:

Bcl-2-associated X protein

Bcl-xL:

B-cell lymphoma-extra-large

ERK:

Extracellular signal-regulated kinase ½

FOXM1:

Forkhead Box M1

H2A.X:

H2A histone family member X

LC3:

Microtubule-associated protein 1A/1B-light chain 3

PARP:

Poly (ADP-ribose) polymerase

RIP3:

Receptor-interacting serine/threonine-protein kinase 3

SRB:

Sulforhodamine B

TNBC:

Triple-negative breast cancer

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Acknowledgements

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (Grant no. 2018R1D1A1B07047758).

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Authors and Affiliations

  1. Department of Pharmacology, College of Medicine and Intractable Disease Research Center, Dongguk University, Gyeongju, 38066, Republic of Korea

    Sushma Maharjan, Yun-Suk Kwon, Min-Gu Lee, Kyu-Shik Lee & Kyung-Soo Nam

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  1. Sushma Maharjan
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  2. Yun-Suk Kwon
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Contributions

K-SN and K-S Lee designed the experiments. K-SN, SM and K-SL analyzed and interpreted data. SM, Y-SK and M-GL performed the experiments and SM and K-SL wrote the manuscript, which was approved by all authors.

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Correspondence to Kyung-Soo Nam.

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Maharjan, S., Kwon, YS., Lee, MG. et al. Cell cycle arrest-mediated cell death by morin in MDA-MB-231 triple-negative breast cancer cells. Pharmacol. Rep 73, 1315–1327 (2021). https://doi.org/10.1007/s43440-021-00272-w

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  • DOI: https://doi.org/10.1007/s43440-021-00272-w

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