Inhibition of human lens epithelial B-3 cell proliferation by adenovirus-mediated transfer of antisense c-myc construct.
OBJECTIVE
To investigate the effects of adenovirus-mediated transfer of antisense c-myc construct on human lens epithelial B-3 (HLE B-3) cell proliferation, apoptosis and cell cycle.


METHODS
HLE B-3 cell cultures were transduced with replication-defective adenovirus bearing either a nuclear-targeted beta-galactosidase (Ad-lacZ) or an antisense c-myc construct (Ad-AS-myc). The presence of beta-galactosidase activity in the transduced cultures was detected by immunohistochemical X-Gal staining, while c-myc mRNA and protein expression levels were evaluated by RT-PCR and Western blot analysis. HLE B-3 cell proliferation within 96 h after the transduction was analyzed by cell counting and MTT colorimetric assay. Apoptosis and cell cycle of the HLE-B3 cells were examined by flow-cytometric analysis.


RESULTS
The mean transduction efficiency was 80% for HLE B-3 cells. Downregulation of c-myc mRNA and protein expression was noticed at 48, 96 and 144 h after the transduction with Ad-AS-myc. Cytostatic effects of Ad-AS-myc in HLE B-3 cells were obvious within 96 h after the transduction. An increased incidence of apoptosis and G1-phase arrest was identified in the Ad-AS-myc-transduced HLE B-3 cells.


CONCLUSIONS
HLE B-3 cells were successfully transduced with adenovirus-mediated antisense c-myc construct. Ad-AS-myc transduction could significantly inhibit cell proliferation and induce cell apoptosis and G1-phase arrest in HLE B-3 cells. It may provide a novel approach for prevention of posterior capsular opacification.
Sun
Jie
J
Shandong Eye Institute & Hospital, 5 Yanerdao Road, Qingdao, 266071, People's Republic of China.
Xie
Lixin
L
Wang
Yao
Y
Liu
Ting
T
D000260:Adenoviruses, Human / Q000235:genetics*; D017209:Apoptosis; D015153:Blotting, Western; D002453:Cell Cycle; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology*; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000502:physiology; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D007908:Lens, Crystalline / Q000166:cytology* / Q000821:virology; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D020319:Oligodeoxyribonucleotides, Antisense; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger