Document Detail

Prolonged down regulation of specific gene expression in nucleus pulposus cell mediated by RNA interference in vitro.
MedLine Citation:
PMID:  16705690     Owner:  NLM     Status:  MEDLINE    
To investigate the efficacies and the longevity of RNA interference in nucleus pulposus cells from rat and human, two reporter luciferase plasmids (Firefly and Renilla) were used. These plasmids were cotransfected with siRNA targeting Firefly luciferase to the nucleus pulposus cells extracted from Sprague Dawley rats and scoliosis patients. The inhibitory effects were evaluated by dual luciferase assay for 3 weeks. Proliferation activity of fibroblast-like cells extracted from the subcutaneous tissue of Sprague Dawley rats and the nucleus pulposus cells were measured by proliferation assay (WST-8 assay) every 2 days after plating. The expression of Firefly luciferase was drastically inhibited both in rats (94.7%) and in humans (93.7%). The inhibitory effects were maintained for 2 weeks and had disappeared completely by 3 weeks. The proliferation activity of nucleus pulposus cells was significantly lower than fibroblast-like cells. We have shown, for the first time, siRNA-mediated gene silencing in rat and human disc cells for a relatively sustained period, probably due to the stability of the nucleus pulposus cells in terms of cell proliferation. The demonstration of this study may allow further exploration of the use of siRNA for scientific research and the treatment of disc degenerative diseases.
Kenichiro Kakutani; Kotaro Nishida; Koki Uno; Toru Takada; Takatoshi Shimomura; Koichiro Maeno; Masahiro Kurosaka; Minoru Doita
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of orthopaedic research : official publication of the Orthopaedic Research Society     Volume:  24     ISSN:  0736-0266     ISO Abbreviation:  J. Orthop. Res.     Publication Date:  2006 Jun 
Date Detail:
Created Date:  2006-05-22     Completed Date:  2006-06-29     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8404726     Medline TA:  J Orthop Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1271-8     Citation Subset:  IM    
Copyright Information:
(c) 2006 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.
Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, Kobe, Japan.
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MeSH Terms
Cell Proliferation
Cells, Cultured
Fibroblasts / cytology,  enzymology*
Green Fluorescent Proteins / genetics,  metabolism
Intervertebral Disk / enzymology*,  pathology
Luciferases / genetics,  metabolism
Microscopy, Fluorescence
RNA Interference / physiology*
RNA, Small Interfering / genetics
RNA-Induced Silencing Complex / genetics
Rats, Sprague-Dawley
Scoliosis / metabolism,  pathology
Reg. No./Substance:
0/RNA, Small Interfering; 0/RNA-Induced Silencing Complex; 147336-22-9/Green Fluorescent Proteins; EC 1.13.12.-/Luciferases

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