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Erschienen in:
01.06.2005 | Laboratory Investigation
Inhibition of human lens epithelial B-3 cell proliferation by adenovirus-mediated transfer of antisense c-myc construct
Erschienen in: Graefe's Archive for Clinical and Experimental Ophthalmology | Ausgabe 6/2005
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Purpose
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 β-galactosidase (Ad-lacZ) or an antisense c-myc construct (Ad-AS-myc). The presence of β-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.