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The synergistic anti-proliferative effect of the combination of diosmin and BEZ-235 (dactolisib) on the HCT-116 colorectal cancer cell line occurs through inhibition of the PI3K/Akt/mTOR/NF-κB axis

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Abstract

One of the most lethal malignancies worldwide is colorectal cancer (CRC). Alterations in various signalling pathways, including PI3K-mTOR and NF-κB, have been reported in CRC with subsequent dysregulation of proliferation, apoptosis, angiogenesis and, questionably, autophagy processes. BEZ-235 (dactolisib) is a dual PI3K-mTOR inhibitor with potent anti-tumour activity. However, the observed toxicity of BEZ-235 necessitated the termination of its clinical trials. Hence, we aimed to evaluate the potential long-lasting anti-carcinogenic effects of adding diosmin (DIO, a natural NF-κB inhibitor) to BEZ-235 in HCT-116 CRC cells. The median inhibitory concentrations (IC50s) of BEZ-235 and/or DIO were evaluated in the HCT-116 CRC cell line. Caspase-3 activity was assessed colorimetrically, and p-Akt, NF-κB, CD1, VEGF and LC3B levels were assessed by ELISA. Additionally, LC3-II and P62 gene expression were assessed using qRT-PCR. The observed CIs (combination indices) and DRIs (dose reduction indices) confirmed the synergistic effect of DIO and BEZ-235. Co-administration of both drugs either in combination-1 (1 μM for BEZ-235, 250 μM for DIO) or in combination-2 (0.51 μM for BEZ-235 + 101.99 μM for DIO) inhibited the PI3K/Akt/mTOR/NF-κB axis, leading to the induction of apoptosis (via active caspase-3), and the inhibition of proliferation marker (CD1), angiogenesis marker (VEGF), autophagy protein (LC3B) and altered effects on LC3-IIandP62 gene expression. Our results reveal the synergistic chemotherapeutic effects of DIO combined with BEZ-235 in the HCT-116 CRC cell line and encourage future preclinical and clinical studies of this combination with reduced BEZ-235 concentrations to avoid its reported toxicity.

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Abbreviations

CD1:

Cyclin D1

CRC:

Colorectal cancer

HIF1α:

Hypoxia-inducible factor-1 alpha

LC3B:

Human microtubule-associated proteins 1A/1B light chain 3 beta

LC3-II:

Microtubule-associated protein 1 light chain 3

mTOR:

Mammalian target of rapamycin

NF-κB:

Nuclear factor-kappa B

p-Akt:

Phospho-Akt

PI3K:

Phosphatidylinositol-3-kinase

STAT-3:

Signal transducer and activator of transcription-3

VEGF:

Vascular endothelial growth factor

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The authors declare that this research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Authors and Affiliations

  1. Pharmacology and Toxicology, Faculty of Pharmacy, Damanhour University, Damanhour, 22514, Egypt

    Maged. W. Helmy, Asser I. Ghoneim, Mohamed A. Katary & Rana K. Elmahdy

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  1. Maged. W. Helmy
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  2. Asser I. Ghoneim
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  3. Mohamed A. Katary
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  4. Rana K. Elmahdy
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Contributions

MWH: Main research idea, experimental design, work supervision, data analysis and presentation, manuscript preparation and revision. AIG: Experimental design, sharing research ideas and work supervision, data analysis and presentation, and manuscript preparation and revision. MAK: Sharing ideas and work supervision, and manuscript preparation. RKE: Experimental design, sharing ideas, performance of the experiments, data analysis and presentation, manuscript preparation.

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Correspondence to Maged. W. Helmy.

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The current research followed accepted principles of ethical and professional conduct according to approval reference number 1216PO3 issued by the Research Ethics Committee of the Faculty of Pharmacy, Damanhour University, regarding originality, risk control, and community service.

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Helmy, M.W., Ghoneim, A.I., Katary, M.A. et al. The synergistic anti-proliferative effect of the combination of diosmin and BEZ-235 (dactolisib) on the HCT-116 colorectal cancer cell line occurs through inhibition of the PI3K/Akt/mTOR/NF-κB axis. Mol Biol Rep 47, 2217–2230 (2020). https://doi.org/10.1007/s11033-020-05327-4

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  • DOI: https://doi.org/10.1007/s11033-020-05327-4

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