Down regulation of the PEDF gene in human lens epithelium cells changed the expression of proteins vimentin and alphaB-crystallin.
OBJECTIVE
To study the relationship of pigment epithelium-derived factor (PEDF) expression with the expression of vimentin and alphaB-crystallin by lens epithelial cells.


METHODS
Lens epithelial cells adhering to anterior capsules taken from young donor eyes aged from 20 to 35 years were cultured and passaged. We designed small interfering RNA (siRNA) constructs to specifically downregulate the expression of PEDF by these primary lens epithelial cells. Quantitative PCR was used to confirm the downregulation of PEDF RNA expression following infection of lens epithelial cells. To determine whether altering the expression of PEDF would effect the expression of vimentin or alphaB-crystallin, we performed western blotting 48 h after expression of the PEDF-directed siRNA.


RESULTS
PEDF RNA expression in the human lens epithelial cells was strongly downregulated by the three separate siRNA constructs. Western blotting revealed that the downregulation of PEDF expression resulted in a concomitant decrease in expression of vimentin and an increase in alphaB-crystallin protein.


CONCLUSIONS
Decreased expression of PEDF in primary human lens epithelial cells resulted in a decrease in the expression of vimentin and the increase of alphaB-crystallin expression, two proteins critical for maintaining lens clarity.
Yang
Jing
J
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Luo
Lixia
L
Liu
Xialin
X
Rosenblatt
Mark I
MI
Qu
Bo
B
Liu
Yuhua
Y
Liu
Yizhi
Y
D000328:Adult; D002478:Cells, Cultured; D015536:Down-Regulation / Q000235:genetics*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D005136:Eye Proteins / Q000235:genetics*; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007908:Lens, Crystalline / Q000166:cytology*; D016086:Lentivirus / Q000235:genetics; D008297:Male; D009414:Nerve Growth Factors / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D015843:Serpins / Q000235:genetics*; D014161:Transduction, Genetic; D014746:Vimentin / Q000378:metabolism*; D055815:Young Adult; D038203:alpha-Crystallin B Chain / Q000378:metabolism*; D000099174:Pigment Epithelium-Derived Factor
D005136:Eye Proteins; D009414:Nerve Growth Factors; D034741:RNA, Small Interfering; D015843:Serpins; D014746:Vimentin; D038203:alpha-Crystallin B Chain; D000099174:Pigment Epithelium-Derived Factor