MicroRNA regulation of the serine synthesis pathway in endocrine-resistant breast cancer cells

in Endocrine-Related Cancer
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Belinda J Petri Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA

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Kellianne M Piell Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA

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Ali E Wilt Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA

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Alexa D Howser Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA

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Laura Winkler Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA

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Mattie R Whitworth Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA

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Bailey L Valdes Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA

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Norman L Lehman Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA
Pathology and Laboratory Medicine, University of Louisville, Louisville, Kentucky, USA
The Brown Cancer Center, University of Louisville, Louisville, Kentucky, USA

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Brian F Clem Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA
The Brown Cancer Center, University of Louisville, Louisville, Kentucky, USA

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https://orcid.org/0000-0002-9082-8914
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Carolyn M Klinge Department of Biochemistry & Molecular Genetics, University of Louisville School of Medicine Louisville, Kentucky, USA
The Brown Cancer Center, University of Louisville, Louisville, Kentucky, USA
Center for Integrative Environmental Health Sciences (CIEHS), University of Louisville, Louisville, Kentucky, USA

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https://orcid.org/0000-0002-3358-4378

Correspondence should be addressed to C M Klinge: carolyn.klinge@louisville.edu
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Despite the successful combination of therapies improving survival of estrogen receptor α (ER+) breast cancer patients with metastatic disease, mechanisms for acquired endocrine resistance remain to be fully elucidated. The RNA binding protein HNRNPA2B1 (A2B1), a reader of N(6)-methyladenosine (m6A) in transcribed RNA, is upregulated in endocrine-resistant, ER+ LCC9 and LY2 cells compared to parental MCF-7 endocrine-sensitive luminal A breast cancer cells. The miRNA-seq transcriptome of MCF-7 cells overexpressing A2B1 identified the serine metabolic processes pathway. Increased expression of two key enzymes in the serine synthesis pathway (SSP), phosphoserine aminotransferase 1 (PSAT1) and phosphoglycerate dehydrogenase (PHGDH), correlates with poor outcomes in ER+ breast patients who received tamoxifen (TAM). We reported that PSAT1 and PHGDH were higher in LCC9 and LY2 cells compared to MCF-7 cells and their knockdown enhanced TAM sensitivity in these-resistant cells. Here we demonstrate that stable, modest overexpression of A2B1 in MCF-7 cells increased PSAT1 and PHGDH and endocrine resistance. We identified four miRNAs downregulated in MCF-7-A2B1 cells that directly target the PSAT1 3′UTR (miR-145-5p and miR-424-5p), and the PHGDH 3′UTR (miR-34b-5p and miR-876-5p) in dual luciferase assays. Lower expression of miR-145-5p and miR-424-5p in LCC9 and ZR-75-1-4-OHT cells correlated with increased PSAT1 and lower expression of miR-34b-5p and miR-876-5p in LCC9 and ZR-75-1-4-OHT cells correlated with increased PHGDH. Transient transfection of these miRNAs restored endocrine-therapy sensitivity in LCC9 and ZR-75-1-4-OHT cells. Overall, our data suggest a role for decreased A2B1-regulated miRNAs in endocrine resistance and upregulation of the SSP to promote tumor progression in ER+ breast cancer.

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