TY - JOUR

T1 - MiR-21 regulates the ACAT1 gene in MCF-7 cells

AU - Chanyshev, M. D.

AU - Razumova, Y. V.

AU - Ovchinnikov, V. Y.

AU - Gulyaeva, L. F.

N1 - Copyright © 2018 Elsevier Inc. All rights reserved.

PY - 2018/9/15

Y1 - 2018/9/15

N2 - Aims: The purpose of the present study was to determine whether miR-21 regulates the human ACAT1 gene. We also assessed whether transfection of MCF-7 cells with miR-21 mimic/inhibitor leads to changes in ACAT1 mRNA/protein levels, cell proliferation rate, or apoptosis. Main methods: Regulation of ACAT1 3′UTR by miR-21 was evaluated using a dual-luciferase reporter assay. The effect of miR-21 on mRNA/protein levels of ACAT1 and PTEN (confirmed as an important target of miR-21 for comparison) was measured by qPCR/western blot analysis and immunostaining. Proliferation rate was determined by cell counting. Percentage of cells undergoing late apoptosis was determined by staining with Hoechst 33342/propidium iodide. Key findings: Dual-luciferase reporter assay confirmed the regulation of ACAT1 3′UTR by miR-21. Furthermore, transfection of MCF-7 cells with miR-21 mimic decreased mRNA and protein levels of ACAT1 and PTEN genes. In contrast, miR-21 inhibition increased the mRNA and protein levels of both genes studied. Finally, we observed an increase in cell proliferation and decrease in the percentage of cells in late apoptosis in MCF-7 cells transfected with miR-21 mimic, whereas transfection with miR-21 inhibitor led to the opposite effect. Significance: Our data confirm the hypothesis that miR-21 regulates the human ACAT1 gene. As the expression of this microRNA is altered in many types of cancers, the discovery of novel targets for miR-21 is of particular interest for diagnosis and treatment.

AB - Aims: The purpose of the present study was to determine whether miR-21 regulates the human ACAT1 gene. We also assessed whether transfection of MCF-7 cells with miR-21 mimic/inhibitor leads to changes in ACAT1 mRNA/protein levels, cell proliferation rate, or apoptosis. Main methods: Regulation of ACAT1 3′UTR by miR-21 was evaluated using a dual-luciferase reporter assay. The effect of miR-21 on mRNA/protein levels of ACAT1 and PTEN (confirmed as an important target of miR-21 for comparison) was measured by qPCR/western blot analysis and immunostaining. Proliferation rate was determined by cell counting. Percentage of cells undergoing late apoptosis was determined by staining with Hoechst 33342/propidium iodide. Key findings: Dual-luciferase reporter assay confirmed the regulation of ACAT1 3′UTR by miR-21. Furthermore, transfection of MCF-7 cells with miR-21 mimic decreased mRNA and protein levels of ACAT1 and PTEN genes. In contrast, miR-21 inhibition increased the mRNA and protein levels of both genes studied. Finally, we observed an increase in cell proliferation and decrease in the percentage of cells in late apoptosis in MCF-7 cells transfected with miR-21 mimic, whereas transfection with miR-21 inhibitor led to the opposite effect. Significance: Our data confirm the hypothesis that miR-21 regulates the human ACAT1 gene. As the expression of this microRNA is altered in many types of cancers, the discovery of novel targets for miR-21 is of particular interest for diagnosis and treatment.

KW - ACAT1

KW - Cancer

KW - MCF-7

KW - miR-21

KW - PTEN

KW - MicroRNAs/genetics

KW - Cell Proliferation

KW - Humans

KW - Gene Expression Regulation, Neoplastic

KW - Biomarkers, Tumor/genetics

KW - MCF-7 Cells

KW - Female

KW - Breast Neoplasms/genetics

KW - Tumor Cells, Cultured

KW - Acetyl-CoA C-Acetyltransferase/genetics

KW - Apoptosis

KW - Cell Movement

UR - http://www.scopus.com/inward/record.url?scp=85051111964&partnerID=8YFLogxK

U2 - 10.1016/j.lfs.2018.08.010

DO - 10.1016/j.lfs.2018.08.010

M3 - Article

C2 - 30092298

AN - SCOPUS:85051111964

VL - 209

SP - 173

EP - 178

JO - Life Sciences

JF - Life Sciences

SN - 0024-3205

ER -