[Effects of small interference RNA targeting connective tissue growth factor on hypoxia-induced collagen synthesis in human lung fibroblasts].
OBJECTIVE
To investigate the effects of small interference RNA (siRNA) targeting connective tissue growth factor (CTGF) on collagen synthesis induced by hypoxia in human lung fibroblasts.


METHODS
CTGF-siRNA expression plasmid was stablely transfected into MRC-5. The transfected cells were cultured under hypoxia condition (37 degrees C, 5% CO2, 1% O2) for 1, 2, 4, 6 and 12 hours respectively. The expression of CTGF and collagen type I was assessed by real time quantitative PCR and Western blot.


RESULTS
Expression of CTGF mRNA was up-regulated as early as 1 hour in control scrambled-siRNA/MRC-5 cells after exposed to hypoxia condition, which remained the elevated level up to 6 hours. Compared with control cells, the expression of CTGF mRNA and protein induced by hypoxia was decreased significantly in CTGF-siRNA/MRC-5 cells. Moreover, the deposition of collagen type I in CTGF-siRNA/MRC-5 cells was also declined.


CONCLUSIONS
The siRNA targeting CTGF gene could efficiently inhibit collagen deposition induced by hypoxia in human lung fibroblasts.
Wang
You-juan
YJ
Laboratory of Cardiovascular Diseases, West China Hospital, Sichuan University, Chengdu 610041, China.
Huang
Yan
Y
Tang
Hui
H
Liu
Xiao-jing
XJ
D015687:Cell Hypoxia; D002478:Cells, Cultured; D024042:Collagen Type I / Q000096:biosynthesis* / Q000235:genetics; D055513:Connective Tissue Growth Factor / Q000235:genetics* / Q000378:metabolism; D015536:Down-Regulation; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008168:Lung / Q000166:cytology*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
C526317:CCN2 protein, human; D024042:Collagen Type I; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D055513:Connective Tissue Growth Factor