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

Research output: Contribution to journalArticle

2 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
Publication statusPublished - Mar 15 2017

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Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Endocrinology

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