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Resistance to the antiproliferative effect induced by a short-chain ceramide is associated with an increase of glucosylceramide synthase, P-glycoprotein, and multidrug-resistance gene-1 in cervical cancer cells

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Abstract

Purpose

Ceramide is glycosylated to glucosylceramide or lactosylceramide, and this glycosylation is a novel multidrug-resistance (MDR) mechanism. In this work, a short-chain ceramide (C6), lactosylceramide (LacCer), and an inhibitor of ceramide glycosylation (d-threo-1-phenyl-2-decanoylamino-3-1-propanol, PDMP) were evaluated on the proliferation of cervical cancer cells. The participation of glucosylceramide synthase (GCS), P-glycoprotein (P-gp), and multidrug-resistance gene-1 (MDR-1) in the resistance to the antiproliferative effect induced by C6 was also evaluated.

Methods

Cell proliferation was determined by crystal violet staining. GCS and MDR-1 mRNA expression was evaluated by real-time RT-PCR assay. GCS and P-gp protein expressions, as well as Rhodamine 123 uptake, which is a functional test for P-gp efflux activity, were determined by flow cytometry.

Results

C6 inhibited proliferation of CaLo and CasKi cells with an IC50 of 2.5 μM; however, 50 % proliferation of ViBo cells was inhibited with 10 μM. LacCer increased the proliferation of all cells. When cells were treated with PDMP plus C6, no additional effect on antiproliferation induced by C6 was observed in CaLo and CasKi cells; however, proliferation diminished in comparison with C6 alone in ViBo cells. C6 increased GCS and MDR-1 expression in all cells, as well as P-gp expression in CasKi cells.

Conclusions

Cells that have more capacity to glycosylate ceramide and express a higher level of GCS, MDR-1, and P-gp, are more resistant to the antiproliferative effect induced by C6.

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Abbreviations

C6:

N-hexanoyl-d-sphingosine

GlcCer:

Glucosylceramide

MDR:

Multidrug resistance

MDR-1:

Multidrug-resistance gene-1

GCS:

Glucosylceramide synthase

P-gp:

P-glycoprotein

HPV:

Human papillomavirus

LacCer:

Lactosylceramide

PDMP:

d-Threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol

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Acknowledgments

We thank Dr. Brianne O’Leary for critical reading of the manuscript. This investigation was financially supported by the CONACyT, Project Number 182341.

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Authors and Affiliations

  1. Laboratorio de Medicina Regenerativa y Estudios en Cáncer, Escuela Superior de Medicina, Instituto Politécnico Nacional, México, México

    Gisela Gutiérrez-Iglesias

  2. Departamento de Biología Celular, Instituto Nacional de Cardiología “Ignacio Chávez”, Juan Badiano No. 1, Colonia Sección 16, Tlalpan, 14080, México, DF, México

    Yamilec Hurtado & Rebeca López-Marure

  3. Laboratorio de Morfología Celular y Molecular, Departamento de Posgrado, Escuela Superior de Medicina, Instituto Politécnico Nacional, México, México

    Icela Palma-Lara

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  1. Gisela Gutiérrez-Iglesias
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  2. Yamilec Hurtado
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  3. Icela Palma-Lara
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  4. Rebeca López-Marure
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Correspondence to Rebeca López-Marure.

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Gutiérrez-Iglesias, G., Hurtado, Y., Palma-Lara, I. et al. Resistance to the antiproliferative effect induced by a short-chain ceramide is associated with an increase of glucosylceramide synthase, P-glycoprotein, and multidrug-resistance gene-1 in cervical cancer cells. Cancer Chemother Pharmacol 74, 809–817 (2014). https://doi.org/10.1007/s00280-014-2552-3

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  • DOI: https://doi.org/10.1007/s00280-014-2552-3

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