Document Detail


Potent siRNA inhibitors of ribonucleotide reductase subunit RRM2 reduce cell proliferation in vitro and in vivo.
MedLine Citation:
PMID:  17404105     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: Ribonucleotide reductase (RR) is a therapeutic target for DNA replication-dependent diseases such as cancer. Here, a potent small interfering RNA (siRNA) duplex against the M2 subunit of RR (RRM2) is developed and shown to reduce the growth potential of cancer cells both in vitro and in vivo. EXPERIMENTAL DESIGN: Three anti-RRM2 siRNAs were identified via computational methods, and the potency of these and additional "tiling" duplexes was analyzed in cultured cells via cotransfections using a RRM2-luciferase fusion construct. Knockdown of RRM2 by the best duplex candidates was confirmed directly by Western blotting. The effect of potent duplexes on cell growth was investigated by a real-time cell electronic sensing assay. Finally, duplex performance was tested in vivo in luciferase-expressing cells via whole animal bioluminescence imaging. RESULTS: Moderate anti-RRM2 effects are observed from the three duplexes identified by computational methods. However, the tiling experiments yielded an extremely potent duplex (siR2B+5). This duplex achieves significant knockdown of RRM2 protein in cultured cells and has pronounced antiproliferative activity. S.c. tumors of cells that had been transfected with siR2B+5 preinjection grew slower than those of control cells. CONCLUSIONS: An anti-RRM2 siRNA duplex is identified that exhibits significant antiproliferative activity in cancer cells of varying human type and species (mouse, rat, monkey); these findings suggest that this duplex is a promising candidate for therapeutic development.
Authors:
Jeremy D Heidel; Joanna Yi-Ching Liu; Yun Yen; Bingsen Zhou; Bret S E Heale; John J Rossi; Derek W Bartlett; Mark E Davis
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Publication Detail:
Type:  In Vitro; Journal Article    
Journal Detail:
Title:  Clinical cancer research : an official journal of the American Association for Cancer Research     Volume:  13     ISSN:  1078-0432     ISO Abbreviation:  Clin. Cancer Res.     Publication Date:  2007 Apr 
Date Detail:
Created Date:  2007-04-03     Completed Date:  2007-06-21     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9502500     Medline TA:  Clin Cancer Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2207-15     Citation Subset:  IM    
Affiliation:
Calando Pharmaceuticals, Inc., California Institute of Technology, Pasadena, California 91107, USA. jheidel@calandopharma.com
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Blotting, Western
Cell Line, Tumor
Cell Proliferation
Gene Therapy / methods*
Humans
Molecular Sequence Data
RNA, Small Interfering / chemical synthesis*,  genetics*
Ribonucleoside Diphosphate Reductase / antagonists & inhibitors*,  genetics*
Sequence Homology, Nucleic Acid
Transfection
Chemical
Reg. No./Substance:
0/RNA, Small Interfering; EC 1.17.4.-/ribonucleotide reductase M2; EC 1.17.4.1/Ribonucleoside Diphosphate Reductase

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


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