[Infection of recombinant adenovirus expressing acidic fibroblasts growth factor enhances rabbit corneal endothelium proliferation].
OBJECTIVE
To transfer the gene encoding mammalian aFGF to rabbit corneal endothelial cell and determine transfection efficiency and the effect of Ad-aFGF on proliferation of RCEC.


METHODS
Replication-defective adenovirus was used to deliver the LacZ reporter gene to RCEC to determine transfection efficiency by X-gal staining. A cDNA encoding mammalian aFGF was cloned into an adenoviral vector that was used to transfect RCEC. The expression of aFGF gene mRNA and protein were demonstrated by Reverse transcription PCR and Western blot respectively. The effect of Ad-aFGF on proliferation of RCEC was determined by MTT assay.


RESULTS
RT-PCR and Western blot from RCEC infected with Ad-aFGF for 48 h detected a specific cDNA amplification and protein expression of aFGF gene, and no positive result in control cells. Consistently, RCEC infected by MOI 20 of Ad-aFGF demonstrated an enhanced RCEC proliferation compared to controls (F = 217.107, P < 0.05).


CONCLUSIONS
Adenoviral-mediated expression of aFGF can stimulate the proliferation of RCEC.
Department of Ophthalmology, First Affiliated Hospital, the Second Military Medical University, Shanghai 200433, China. linliu@public6.sta.net.cn
Lin
L
Liu
Yong
Y
Wang
Xin
X
Pan
Ming
M
Zhong
Xin-Fu
XF
Wen
Zhong-Tian
ZT
Qi
Zhi-Yong
ZY
Liu
Hong
H
Liu
Wei-Qiang
WQ
Tang
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000235:genetics; D004728:Endothelium, Corneal / Q000166:cytology / Q000378:metabolism*; D005260:Female; D016220:Fibroblast Growth Factor 1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008297:Male; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D004274:DNA, Recombinant; D012333:RNA, Messenger; D016220:Fibroblast Growth Factor 1