Document Detail

A novel microRNA mmu-miR-466h affects apoptosis regulation in mammalian cells.
MedLine Citation:
PMID:  21337326     Owner:  NLM     Status:  MEDLINE    
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.
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.
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
CHO Cells
Caspase 3 / metabolism
Caspase 7 / metabolism
Cell Survival
Culture Media / chemistry
Gene Expression Profiling
Gene Expression Regulation*
MicroRNAs / metabolism*
Reverse Transcriptase Polymerase Chain Reaction
Grant Support
Z01 DK070010-02/DK/NIDDK NIH HHS
Reg. No./Substance:
0/Culture Media; 0/MIRN191 microRNA, mouse; 0/MicroRNAs; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 7

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