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This Article Full Text Full Text (PDF) All Versions of this Article: iovs.08-2060v1 49/11/5111    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Sharma, A. Articles by Kenney, M. C. Search for Related Content PubMed PubMed Citation Articles by Sharma, A. Articles by Kenney, M. C.

Effects of Benzo(e)Pyrene, a Toxic Component of Cigarette Smoke, on Human Retinal Pigment Epithelial Cells In Vitro

¹From the Department of Ophthalmology, School of Medicine, University of California, Irvine, California; the ²Department of Ophthalmology and Visual Sciences, University of Wisconsin, Madison, Wisconsin; and the ³Departamento de Oftalmologia, Fundacion VER, Cordoba, Argentina.

PURPOSE. To better understand the cellular and molecular basis for the epidemiologic association between cigarette smoke and age-related macular degeneration (AMD), the authors examined the effects of Benzo(e)Pyrene (B(e)P), a toxic element in cigarette smoke, on human retinal pigment epithelial cells (ARPE-19).

METHODS. ARPE-19 cells were cultured in Dulbecco modified Eagle medium containing 10% fetal bovine serum. Cells were treated for 24 hours with 1000 µM, 400 µM, 200 µM, and 100 µM B(e)P. Cell viability was determined by a trypan blue dye-exclusion assay. Activities of caspase-3/7, caspase-8, caspase-9, and caspase-12 were measured by a fluorescence image scanner, and DNA laddering was evaluated by electrophoresis on 3% agarose gel.

RESULTS. The mean percentage of cell viabilities of ARPE-19 cells was decreased in a dose-dependent manner after exposure to B(e)P at the higher concentrations of 1000 µM (20.0 ± 0.4; P < 0.001), 400 µM (35.6 ± 6.4; P < 0.001), and 200 µM (58.7 ± 2.3; P < 0.001) but not at 100 µM (95.9 ± 0.7; P > 0.05) compared with the equivalent dimethyl sulfoxide (DMSO)-treated control cultures. There were significant increases in caspase-3/7, -8, -9, and -12 activities compared with the DMSO-treated controls (P < 0.001). DNA laddering revealed bands at 200-bp intervals.

CONCLUSIONS. These results show that B(e)P is a toxicant to human retinal pigment epithelial cells in vitro. It causes cell death and induces apoptosis by the involvement of multiple caspase pathways.