Clin Exp Reprod Med.  2012 Jun;39(2):87-93. 10.5653/cerm.2012.39.2.87.

Lin28 regulates the expression of neuropeptide Y receptors and oocyte-specific homeobox genes in mouse embryonic stem cells

• Park GT
• Seo YM
• Lee SY
• Lee KA

• KMID: 1743696
• DOI: http://doi.org/10.5653/cerm.2012.39.2.87

Abstract

OBJECTIVE
Lin28 has been known to control the proliferation and pluripotency of embryonic stem cells. The purpose of this study was to determine the downstream effectors of Lin28 in mouse embryonic stem cells (mESCs) by RNA interference and microarray analysis.
METHODS
The control siRNA and Lin28 siRNA (Dharmacon) were transfected into mESCs. Total RNA was prepared from each type of transfected mESC and subjected to reverse transcription-polymerase chain reaction (RT-PCR) analysis to confirm the downregulation of Lin28. The RNAs were labeled and hybridized with an Affymetrix Gene-Chip Mouse Genome 430 2.0 array. The data analysis was accomplished by GenPlex 3.0 software. The expression levels of selected genes were confirmed by quantitative real-time RT-PCR.
RESULTS
According to the statistical analysis of the cDNA microarray, a total of 500 genes were altered in Lin28-downregulated mESCs (up-regulated, 384; down-regulated, 116). After differentially expressed gene filtering, 31 genes were selected as candidate genes regulated by Lin28 downregulation. Among them, neuropeptide Y5 receptor and oocyte-specific homeobox 5 genes were significantly upregulated in Lin28-downregulated mESCs. We also showed that the families of neuropeptide Y receptor (Npyr) and oocyte-specific homeobox (Obox) genes were upregulated by downregulation of Lin28.
CONCLUSION
Based on the results of this study, we suggest that Lin28 controls the characteristics of mESCs through the regulation of effectors such as the Npyr and Obox families.

Keyword

Lin28; Embryonic stem cells; RNA interference; Microarray; Neuropeptide Y receptors; Oocyte-specific homeobox

MeSH Terms

Animals
Chimera
Down-Regulation
Embryonic Stem Cells
Genes, Homeobox
Genome
Humans
Mice
Neuropeptide Y
Neuropeptides
Oligonucleotide Array Sequence Analysis
Receptors, Neuropeptide Y
RNA
RNA Interference
RNA, Small Interfering
Statistics as Topic
Neuropeptide Y
Neuropeptides
RNA
RNA, Small Interfering
Receptors, Neuropeptide Y