Effects of Benzo(e)Pyrene, a toxic component of cigarette smoke, on human retinal pigment epithelial cells in vitro.

Abstract

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 microM, 400 microM, 200 microM, and 100 microM 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 microM (20.0 +/- 0.4; P < 0.001), 400 microM (35.6 +/- 6.4; P < 0.001), and 200 microM (58.7 +/- 2.3; P < 0.001) but not at 100 microM (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.

• Authors: Ashish Sharma, Aneesh Neekhra, Ana L Gramajo, Jayaprakash Patil, Marilyn Chwa, Baruch D Kuppermann, M Cristina Kenney
• Journal: Investigative ophthalmology & visual science (Invest Ophthalmol Vis Sci) Vol. 49 Issue 11 Pg. 5111-7 (Nov 2008) ISSN: 1552-5783 [Electronic] United States
• PMID: 18586875 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Benzopyrenes
• Tobacco Smoke Pollution
• benzo(e)pyrene
• DNA
• Caspases

Topics

• Apoptosis
• Benzopyrenes (adverse effects)
• Caspases (metabolism)
• Cell Survival
• Cells, Cultured
• DNA (drug effects)
• DNA Fragmentation
• Electrophoresis, Agar Gel
• Humans
• Macular Degeneration (etiology, genetics, pathology)
• Pigment Epithelium of Eye (drug effects, pathology)
• Risk Factors
• Tobacco Smoke Pollution (analysis)