Abstract
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.
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Acknowledgements
This work was supported by a grant from the Shandong Natural Science Foundation (Y2003C04), PR China. The authors thank Dr. Youhai Chen, University of Pennsylvania, for kindly providing the HLE B-3 cell line, Drs. Chen Lin and Qimin Zhan, Chinese Academy of Medical Sciences Institute of Oncology, for kindly providing the Ad-lacZ and Ad-AS-myc, and Ms. Ping Lin for her editorial assistance.
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Sun, J., Xie, L., Wang, Y. et al. Inhibition of human lens epithelial B-3 cell proliferation by adenovirus-mediated transfer of antisense c-myc construct. Graefe's Arch Clin Exp Ophthalmol 243, 601–606 (2005). https://doi.org/10.1007/s00417-004-1095-4
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DOI: https://doi.org/10.1007/s00417-004-1095-4