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Effects of miR-21 downregulation and silibinin treatment in breast cancer cell lines

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Abstract

Silibinin is a natural polyphenol with high antioxidant and anticancer properties, which causes cell cycle arrest and apoptosis in most cancer cell types including breast cancer, but the in-line mechanisms, are still unknown. Silibinin significantly downregulated oncomiR miR-21 expression in breast cancer cells. Here the effect of anti-miR-21 on cell viability, apoptotic induction, cell cycle distribution, and the expression levels of downstream targets of miR-21 were investigated in MCF-7 and T47D cells. MiR-21 mimic transfection was also applied in silibinin treated samples to evaluate functional role of miR-21downregulation on silibinin effects. It was found that after anti-miR-21 transfection, no significant changes were detected in cell viability, apoptosis (except early apoptosis), and cell cycle in MCF-7 and T47D cells. Compared to silibinin, miR-21 mimic transfection in combination with silibinin caused a slight modulation in some of the examined silibinin effects including apoptosis, Bcl2 mRNA and PTEN mRNA and protein levels. Silibinin slightly changed luciferase activity from reporters containing the miR-21 recognition elements from PTEN-3′UTR and Bcl2-3′UTR in both cell lines. Together these data demonstrated negligible cancer-progression impact of miR-21 and limited roles of miR-21 downregulation in examined silibinin effects, and strengthened the anti-cancer pathways of silibinin other than miR-21downregulation in MCF-7 and T47D cells.

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Abbreviations

ERa+ :

Estrogen receptor alpha positive

ERa− :

Estrogen receptor alpha negative

PTEN:

Phosphatase and tensin homolog

SNORD-47:

Small nucleolar RNA 47

PDCD4:

Programmed cell death protein 4

TM1:

Tropomyosine 1

PMSF:

Phenyl methyl sulfonyl fluoride

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Acknowledgements

We would like to express our thanks to Prof. Elaheh Elahi for providing psiCHECK™-2 vector, Dr. Ehsan Arefian, for providing dual-luciferase activity reporter assay kit, Mr Babak Saffari, PhD student for constructing psiCheck-2/PTEN-3′UTR and psiCheck-2/Bcl2-3′UTR vectors, Miss Mahfam Shariati, MSc student for vector extraction and Miss Mehraban Mirrahimi, MSc student for bacterial culture, in Tehran University which help us after manuscript revision.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. The authors greatly acknowledge the financial supports of Science and Technology Park, University of Tehran, Tehran, Iran.

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Authors and Affiliations

  1. Department of Cellular and Molecular Biology, School of Biology, College of Science, University of Tehran, P.O. Box: 14155-6455, Tehran, Iran

    Zohreh Jahanafrooz & Nasrin Motamed

  2. Department of Biotechnology, College of Science, University of Tehran, P.O. Box: 14155-6455, Tehran, Iran

    Behnaz Bakhshandeh

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  1. Zohreh Jahanafrooz
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  2. Nasrin Motamed
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Correspondence to Nasrin Motamed or Behnaz Bakhshandeh.

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Jahanafrooz, Z., Motamed, N. & Bakhshandeh, B. Effects of miR-21 downregulation and silibinin treatment in breast cancer cell lines. Cytotechnology 69, 667–680 (2017). https://doi.org/10.1007/s10616-017-0076-5

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  • DOI: https://doi.org/10.1007/s10616-017-0076-5

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