Regulation of miR-29b-1/a transcription and identification of target mRNAs in CHO-K1 cells

Penn Muluhngwi, Kirsten Richardson, Joshua Napier, Eric C. Rouchka, Justin L. Mott, Carolyn M. Klinge

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6 Scopus citations

Abstract

miR-29b and miR-29a transcript levels were reported to increase in exponentially growing CHO-K1 cells. Here, we examine the regulation of miR-29b-1/a in CHO-K1 cells. We observed that 4-hydroxytamoxifen (4-OHT) increased pri-miR-29b-1 and pri-miR-29a transcription in CHO-K1 cells by activating endogenous estrogen receptor α (ERα). DICER, an established, bona fide target of miR-29b-1/a, was shown to be regulated by 4-OHT in CHO-K1 cells. We showed that miR-29b-1 and miR-29a serve a repressive role in cell proliferation, migration, invasion, and colony formation in CHO-K1 cells. To identify other targets of miR-29b-1 and miR-29a, RNA sequencing was performed by transfecting cells with anti-miR-29a, which inhibits both miR-29a and miR-29b-1, pre-miR-29b-1, and/or pre-miR-29a. In silico network analysis in MetaCore™ identified common and unique putative gene targets of miR-29b-1 and miR-29a. Pathway analysis of identified putative miR-29 targets were related to cell adhesion, cytoskeletal remodeling, and development. Further inquiry revealed regulation of pathways mediating responses to growth factor stimulus and cell cycle regulation.

Original languageEnglish (US)
Pages (from-to)38-47
Number of pages10
JournalMolecular and Cellular Endocrinology
Volume444
DOIs
StatePublished - Mar 15 2017
Externally publishedYes

Bibliographical note

Funding Information:
This work was supported in part by National Institutes of Health R01 CA138410 to C.M.K. and grants from the University of Louisville (UofL) School of Medicine and the UofL Center for Genetics and Molecular Medicine (CGeMM) Next Generation Pilot Grant to C.M.K. Bioinformatics support for this work by E.C.R. was provided by National Institutes of Health grants P20GM103436 (Nigel Cooper, PI). J.N. was supported by National Institutes of Health T35 DK072923 (to C.M.K). We thank Dr. Marsha Cole for the use of her microscope.

Publisher Copyright:
© 2017 Elsevier B.V.

Keywords

  • 4-Hydroxytamoxifen
  • CHO-K1
  • DICER1
  • ERα
  • miRNAs
  • RNA seq

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