Atorvastatin attenuation of ABCB1 expression is mediated by microRNA miR-491-3p in Caco-2 cells

Alice C. Rodrigues, Elida Adalgisa Neri, Sidney Veríssimo-Filho, Nancy Amaral Rebouças, Rosario D C Hirata, Aiming Yu

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Aim Atorvastatin, a HMG-CoA reductase inhibitor, used in the treatment of hypercholesterolemia, has been previously shown to regulate ABCB1 expression in vivo and in vitro. We hypothesized that the statin could regulate gene expression of ABCB1 transporter via microRNAs. Methods Expression of microRNAs and ABCB1 mRNA was examined in atorvastatin-treated and control cells using real-time PCR. miR-491-3P mimic and inhibitor were transfected in Caco-2 and ABCB1 expression was monitored by western blot and real-time PCR. Results In HepG2 cells, none of the microRNAs predicted to target ABCB1 3′UTR was regulated by atorvastatin treatment. In agreement with this, ABCB1 3′UTR activity was not modulated in HepG-2 cells after 48 h-treatment as measured by luciferase assay. In Caco-2 cells, atorvastatin treatment provoked a decrease in luciferase activity and, accordingly, miR-491-3p was upregulated about 2.7 times after 48 h-statin treatment. Luciferase analysis of miR-491-3p with a mimetic or inhibitor of miR-491-3p revealed that this microRNA could target ABCB1 3′UTR, as after miR-491-3p inhibition, ABCB1 levels were increased by two-fold, and miR-491-3p superexpression decreased ABCB1 3′UTR activity. Finally, functional analysis revealed that treatment with miR-491-3p inhibitor could reverses atorvastatin attenuation of ABCB1 (Pg-p) protein levels. Conclusion Our results suggest atorvastatin control ABCB1 expression via miR-491-3p in Caco-2 cells. This finding may be an important mechanism of statin drug–drug interaction, since common concomitant drugs used in the prevention of cardiovascular diseases are ABCB1 substrates.

Original languageEnglish (US)
Pages (from-to)431-436
Number of pages6
JournalEuropean Journal of Pharmaceutical Sciences
Volume93
DOIs
StatePublished - Oct 10 2016

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Caco-2 Cells
MicroRNAs
Hydroxymethylglutaryl-CoA Reductase Inhibitors
Luciferases
Real-Time Polymerase Chain Reaction
Hep G2 Cells
Hypercholesterolemia
Atorvastatin Calcium
Cardiovascular Diseases
Western Blotting
Gene Expression
Messenger RNA
Pharmaceutical Preparations
Proteins

Keywords

  • ABCB1
  • Efflux drug transporters
  • HMG-CoA inhibitors
  • MicroRNAs

ASJC Scopus subject areas

  • Pharmaceutical Science

Cite this

Atorvastatin attenuation of ABCB1 expression is mediated by microRNA miR-491-3p in Caco-2 cells. / Rodrigues, Alice C.; Neri, Elida Adalgisa; Veríssimo-Filho, Sidney; Rebouças, Nancy Amaral; Hirata, Rosario D C; Yu, Aiming.

In: European Journal of Pharmaceutical Sciences, Vol. 93, 10.10.2016, p. 431-436.

Research output: Contribution to journalArticle

Rodrigues, Alice C. ; Neri, Elida Adalgisa ; Veríssimo-Filho, Sidney ; Rebouças, Nancy Amaral ; Hirata, Rosario D C ; Yu, Aiming. / Atorvastatin attenuation of ABCB1 expression is mediated by microRNA miR-491-3p in Caco-2 cells. In: European Journal of Pharmaceutical Sciences. 2016 ; Vol. 93. pp. 431-436.
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T1 - Atorvastatin attenuation of ABCB1 expression is mediated by microRNA miR-491-3p in Caco-2 cells

AU - Rodrigues, Alice C.

AU - Neri, Elida Adalgisa

AU - Veríssimo-Filho, Sidney

AU - Rebouças, Nancy Amaral

AU - Hirata, Rosario D C

AU - Yu, Aiming

PY - 2016/10/10

Y1 - 2016/10/10

N2 - Aim Atorvastatin, a HMG-CoA reductase inhibitor, used in the treatment of hypercholesterolemia, has been previously shown to regulate ABCB1 expression in vivo and in vitro. We hypothesized that the statin could regulate gene expression of ABCB1 transporter via microRNAs. Methods Expression of microRNAs and ABCB1 mRNA was examined in atorvastatin-treated and control cells using real-time PCR. miR-491-3P mimic and inhibitor were transfected in Caco-2 and ABCB1 expression was monitored by western blot and real-time PCR. Results In HepG2 cells, none of the microRNAs predicted to target ABCB1 3′UTR was regulated by atorvastatin treatment. In agreement with this, ABCB1 3′UTR activity was not modulated in HepG-2 cells after 48 h-treatment as measured by luciferase assay. In Caco-2 cells, atorvastatin treatment provoked a decrease in luciferase activity and, accordingly, miR-491-3p was upregulated about 2.7 times after 48 h-statin treatment. Luciferase analysis of miR-491-3p with a mimetic or inhibitor of miR-491-3p revealed that this microRNA could target ABCB1 3′UTR, as after miR-491-3p inhibition, ABCB1 levels were increased by two-fold, and miR-491-3p superexpression decreased ABCB1 3′UTR activity. Finally, functional analysis revealed that treatment with miR-491-3p inhibitor could reverses atorvastatin attenuation of ABCB1 (Pg-p) protein levels. Conclusion Our results suggest atorvastatin control ABCB1 expression via miR-491-3p in Caco-2 cells. This finding may be an important mechanism of statin drug–drug interaction, since common concomitant drugs used in the prevention of cardiovascular diseases are ABCB1 substrates.

AB - Aim Atorvastatin, a HMG-CoA reductase inhibitor, used in the treatment of hypercholesterolemia, has been previously shown to regulate ABCB1 expression in vivo and in vitro. We hypothesized that the statin could regulate gene expression of ABCB1 transporter via microRNAs. Methods Expression of microRNAs and ABCB1 mRNA was examined in atorvastatin-treated and control cells using real-time PCR. miR-491-3P mimic and inhibitor were transfected in Caco-2 and ABCB1 expression was monitored by western blot and real-time PCR. Results In HepG2 cells, none of the microRNAs predicted to target ABCB1 3′UTR was regulated by atorvastatin treatment. In agreement with this, ABCB1 3′UTR activity was not modulated in HepG-2 cells after 48 h-treatment as measured by luciferase assay. In Caco-2 cells, atorvastatin treatment provoked a decrease in luciferase activity and, accordingly, miR-491-3p was upregulated about 2.7 times after 48 h-statin treatment. Luciferase analysis of miR-491-3p with a mimetic or inhibitor of miR-491-3p revealed that this microRNA could target ABCB1 3′UTR, as after miR-491-3p inhibition, ABCB1 levels were increased by two-fold, and miR-491-3p superexpression decreased ABCB1 3′UTR activity. Finally, functional analysis revealed that treatment with miR-491-3p inhibitor could reverses atorvastatin attenuation of ABCB1 (Pg-p) protein levels. Conclusion Our results suggest atorvastatin control ABCB1 expression via miR-491-3p in Caco-2 cells. This finding may be an important mechanism of statin drug–drug interaction, since common concomitant drugs used in the prevention of cardiovascular diseases are ABCB1 substrates.

KW - ABCB1

KW - Efflux drug transporters

KW - HMG-CoA inhibitors

KW - MicroRNAs

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