Document Detail


A novel microRNA mmu-miR-466h affects apoptosis regulation in mammalian cells.
MedLine Citation:
PMID:  21337326     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This study determined the changes in microRNA (miRs) expression in mammalian Chinese hamster ovary (CHO) cells undergoing apoptosis induced by exposing the cells to nutrient-depleted media. The apoptosis onset was confirmed by reduced cell viability and Caspase-3/7 activation. Microarray comparison of known mouse and rat miRs in CHO cells exposed to fresh or depleted media revealed up-regulation of the mouse miR-297-669 cluster in CHO cells subjected to depleted media. The mmu-miR-466h was chosen for further analysis as the member of this cluster with the highest overexpression and its up-regulation in depleted media was confirmed with qRT-PCR. Since miRs suppress mRNA translation, we hypothesized that up-regulated mmu-miR-466h inhibits anti-apoptotic genes and induces apoptosis. A combination of bioinformatics and experimental tools was used to predict and verify mmu-miR-466h anti-apoptotic targets. 8708 predicted targets were obtained from miRecords database and narrowed to 38 anti-apoptotic genes with DAVID NCBI annotation tool. Several genes were selected from this anti-apoptotic subset based on nucleotide pairing complimentarity between the mmu-miR-466h seed region and 3' UTR of the target mRNAs. The qRT-PCR analysis revealed reduced mRNA levels of bcl2l2, dad1, birc6, stat5a, and smo genes in CHO cells exposed to depleted media. The inhibition of the mmu-miR-466h increased the expression levels of those genes and resulted in increased cell viability and decreased Caspase-3/7 activation. The up-regulation of mmu-miR-466h in response to nutrients depletion causes the inhibition of several anti-apoptotic genes in unison. This suggests the pro-apoptotic role of mmu-miR-466h and its capability to modulate the apoptotic pathway in mammalian cells.
Authors:
Aliaksandr Druz; Chia Chu; Brian Majors; Rodell Santuary; Michael Betenbaugh; Joseph Shiloach
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Intramural     Date:  2011-03-11
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  108     ISSN:  1097-0290     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2011 Jul 
Date Detail:
Created Date:  2011-05-19     Completed Date:  2011-08-30     Revised Date:  2013-06-30    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1651-61     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Wiley Periodicals, Inc.
Affiliation:
Biotechnology Core Laboratory National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 9000 Rockville Pike Bldg 14A Rm 176, Bethesda, Maryland 20892, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis*
CHO Cells
Caspase 3 / metabolism
Caspase 7 / metabolism
Cell Survival
Cricetinae
Cricetulus
Culture Media / chemistry
Food
Gene Expression Profiling
Gene Expression Regulation*
Mice
MicroRNAs / metabolism*
Rats
Reverse Transcriptase Polymerase Chain Reaction
Grant Support
ID/Acronym/Agency:
Z01 DK070010-02/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Culture Media; 0/MIRN191 microRNA, mouse; 0/MicroRNAs; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 7
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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