Nanog regulates molecules involved in stemness and cell cycle-signaling pathway for maintenance of pluripotency of P19 embryonal carcinoma stem cells

Seung Cheol Choi, Ji Hyun Choi, Chi Yeon Park, Chul Min Ahn, Soon Jun Hong, Do-Sun Lim

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

To identify potential downstream targets of Nanog, a key transcription factor in the maintenance of pluripotency of embryonic stem (ES) and embryonal carcinoma (EC) cells, global gene expression profiles in Nanog small interfering RNA (siRNA)-transfected P19 EC stem cells were performed using cDNA, 60-mer, and 30-mer microarray platforms. The putative Nanog target genes identified by Nanog silencing were verified using reverse transcription-polymerase chain reaction after Nanog overexpression. Downregulation of Nanog in P19 cells resulted in reduction of pluripotency markers, such as Fgf4, Klf2, Mtf2, Oct-4, Rex1, Sox1, Yes, and Zfp143, whereas overexpression of Nanog in P19 cells reversely upregulated their expression. However, expressions of pluripotency markers Cripto, germ cell nuclear factor, Sox2, and Zfp57 as well as leukemia inhibitory factor (LIF)/Stat3 pathway molecules LIF, IL6st, and Stat3 were not affected after 48h transfection with Nanog siRNA or construct. Nanog silencing also downregulated expression of molecules involved in the p53- and cell cycle-signaling pathway (Atf3, Jdp2, Cul3, Hist1hic, and Bcl6), whereas expression of E2f1, Tob1, Lyn, and Smarcc1 was upregulated by Nanog silencing. Expressions of cyclins D1, D2, D3, and E1 as well as cyclin-dependent kinase (Cdk) 1 and Cdk6 were downregulated by Nanog silencing in P19 cells, whereas Nanog overexpression reversely increased their expressions. Taken together, examination of global transcriptional changes after Nanog silencing followed by verification by Nanog overexpression has revealed new molecules involved in the maintenance of self-renewal and in the regulation of the p53- and cell cycle-pathway of P19 cells.

Original languageEnglish
Pages (from-to)3678-3692
Number of pages15
JournalJournal of Cellular Physiology
Volume227
Issue number11
DOIs
Publication statusPublished - 2012 Nov 1

Fingerprint

Embryonal Carcinoma Stem Cells
Stem cells
Leukemia Inhibitory Factor
Cell Cycle
Cells
Maintenance
Nuclear Receptor Subfamily 6, Group A, Member 1
Small Interfering RNA
Molecules
Down-Regulation
CDC2 Protein Kinase
Polymerase chain reaction
Cyclin D1
Cyclin D2
Transcription
Microarrays
Gene expression
Transcription Factors
Complementary DNA
Genes

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Nanog regulates molecules involved in stemness and cell cycle-signaling pathway for maintenance of pluripotency of P19 embryonal carcinoma stem cells. / Choi, Seung Cheol; Choi, Ji Hyun; Park, Chi Yeon; Ahn, Chul Min; Hong, Soon Jun; Lim, Do-Sun.

In: Journal of Cellular Physiology, Vol. 227, No. 11, 01.11.2012, p. 3678-3692.

Research output: Contribution to journalArticle

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