[Influence on human keratocyte proliferation by cationic liposome mediated transfer of p21WAF1].

Objective: The aim of this study is to investigate the safety and efficiency of transducing human keratocytes with beta-galactosidase gene through cationic liposome and evaluate the influence of p21WAF1 gene transduction mediated by cationic liposome on proliferation of keratocyte.
Methods: (1) Cationic liposome mediated plasmid DNA bearing beta-galactosidase gene was transferred into cultured human keratocyte, and transduction efficiency was evaluated by enzymatic color reaction using X-gal as a substrate. (2) By immunohistochemistry and MTT, the effect of p21WAF1 gene transduction mediated by cationic liposome on cultured keratocyte proliferation was evaluated. (3) Cellular viability was detected by 0.5% trypan blue dye.
Results: (1) Lipofectaminetrade mark reagent could mediate a foreign gene into a cultured human keratocyte, and the highest transduction efficiency was 40.5% when DNA/lipofectamine ratio was 1:8 and the transduction time was 24 hr. (2) p21WAF1 gene transduction mediated by lipofectaminetrade mark reagent could increase the p21 protein expression in the keratocyte, and inhibit the keratocyte proliferation during 2 - 10 days after transduction. And rates of cellular viability in all groups were greater than 90% by trypan blue dye examination.
Conclusion: Cationic liposome can mediate foreign gene into cultured human keratocytes safely and effectively; p21WAF1 gene transduction mediated by cationic liposome can inhibit the proliferation of human keratocyte.