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Treatment of LS174T colorectal cancer stem-like cells with n-3 PUFAs induces growth suppression through inhibition of survivin expression and induction of caspase-3 activation

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Abstract

Purpose

Colorectal cancer stem cells (CCSCs) are thought to contribute to tumor initiation, progression, metastasis, chemo-resistance and therapy failure. Therefore, assessment of the effectiveness of agents with anti-proliferative activities against CCSCs is warranted. Several studies have shown that different tumorigenic steps, ranging from initiation to metastasis, can be affected by n-3 polyunsaturated fatty acids (PUFAs). Here, we evaluated the effects of the PUFA components docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), alone or in combination, on LS174T cells that serve as a model for colorectal cancer initiating cells with stem cell-like properties.

Methods

LS174T cells were treated with 50, 100 and 150 μM DHA and EPA, or equal mixtures of DHA/EPA (i.e., 25/25, 50/50 and 75/75 μM), after which cell number, viability, growth inhibition, survivin expression, caspase-3 activation and apoptotic rate were evaluated.

Results

We found that treatment of LS174T cells with increasing PUFA concentrations significantly increased growth inhibition in a dose- and time-dependent manner. After a 72 h treatment with 150 μM DHA and EPA, or their combination (75/75 μM), growth rates were inhibited by 80.3 ± 5.5 %, 79.3 ± 5 % and 71.1 ± 1 %, respectively, compared to untreated cells. We also found that treatment for 48 h with 100 μM DHA and EPA, or their combination (50/50 μM), resulted in 2.9-, 3- and 2.6-fold increases in caspase-3 activation, as well as 54, 62.4 and 100 % decreases in survivin mRNA expression levels, respectively, compared to untreated cells. Low survivin mRNA levels combined with high caspase-3 activity levels were found to correlate with a higher growth inhibition in PUFA-treated cells. DHA appears to be a more potent growth inhibitor than EPA and the DHA/EPA combination. An increase in the number of apoptotic cells (early + late), ranging from 12.9 to 44.7 %, was observed with increasing DHA doses.

Conclusion

From our data we conclude that PUFAs induce growth inhibition via targeting survivin expression in LS174T cells, which serve as a model for CCSCs.

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Acknowledgments

The authors express their great appreciation for the financial support received for this work from the Urmia University, Iran.

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Authors and Affiliations

  1. Department of Histology and Embryology, Faculty of Science, Urmia University, Urmia, Iran

    Mohammad Reza Sam, Parinaz Ahangar & Vahid Nejati

  2. Department of Cellular and Molecular Biotechnology, Institute of Biotechnology, Urmia University, Urmia, Iran, P.O. Box: 165

    Mohammad Reza Sam & Parinaz Ahangar

  3. Royan Stem Cell Technology Company, West Azerbaijan Cord Blood Bank, Urmia, Iran

    Mohammad Reza Sam

  4. Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

    Reza Habibian

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  1. Mohammad Reza Sam
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Correspondence to Mohammad Reza Sam.

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Sam, M.R., Ahangar, P., Nejati, V. et al. Treatment of LS174T colorectal cancer stem-like cells with n-3 PUFAs induces growth suppression through inhibition of survivin expression and induction of caspase-3 activation. Cell Oncol. 39, 69–77 (2016). https://doi.org/10.1007/s13402-015-0254-4

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