Patulin causes DNA damage leading to cell cycle arrest and apoptosis through modulation of Bax, p(53) and p(21/WAF1) proteins in skin of mice.
Patulin (PAT), a mycotoxin found in apples, grapes, oranges, pear and peaches, is a potent genotoxic compound. WHO has highlighted the need for the study of cutaneous toxicity of PAT as manual labour is employed during pre and post harvest stages, thereby causing direct exposure to skin. In the present study cutaneous toxicity of PAT was evaluated following topical application to Swiss Albino mice. Dermal exposure of PAT, to mice for 4 h resulted in a dose (40-160 mug/animal) and time (up to 6 h) dependent enhancement of ornithine decarboxylase (ODC), a marker enzyme of cell proliferation. The ODC activity was found to be normal after 12 and 24 h treatment of patulin. Topical application of PAT (160 mug/100 mul acetone) for 24-72 h caused (a) DNA damage in skin cells showing significant increase (34-63%) in olive tail moment, a parameter of Comet assay (b) significant G 1 and S-phase arrest along with induction of apoptosis (2.8-10 folds) as shown by annexin V and PI staining assay through flow cytometer. Moreover PAT leads to over expression of p(21/WAF1) (3.6-3.9 fold), pro apoptotic protein Bax (1.3-2.6) and tumor suppressor wild type p(53) (2.8-3.9 fold) protein. It was also shown that PAT induced apoptosis was mediated through mitochondrial intrinsic pathway as revealed through the release of cytochrome C protein in cytosol leading to enhancement of caspase-3 activity in skin cells of mice. These results suggest that PAT has a potential to induce DNA damage leading to p(53) mediated cell cycle arrest along with intrinsic pathway mediated apoptosis that may also be correlated with enhanced polyamine production as evident by induction of ODC activity, which may have dermal toxicological implications.
Saxena
Neha
N
Food Toxicology Division, Indian Institute of Toxicology Research (formerly: Industrial Toxicology Research Centre), Council of Scientific and Industrial Research, Mahatma Gandhi Marg, P.O. Box #80, Lucknow-226001, India.
Ansari
Kausar M
KM
Kumar
Rahul
R
Dhawan
Alok
A
Dwivedi
Premendra D
PD
Das
Mukul
M
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000096:biosynthesis / Q000235:genetics; D002453:Cell Cycle / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D020552:Comet Assay; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics*; D045304:Cytochromes c / Q000096:biosynthesis / Q000235:genetics; D004249:DNA Damage / Q000187:drug effects*; D016158:Genes, p53 / Q000187:drug effects*; D015151:Immunoblotting; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009153:Mutagens / Q000633:toxicity*; D009955:Ornithine Decarboxylase / Q000096:biosynthesis / Q000235:genetics; D010365:Patulin / Q000633:toxicity*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D012879:Skin Physiological Phenomena / Q000235:genetics*; D051028:bcl-2-Associated X Protein / Q000235:genetics*
C493957:Cdkn1a protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D009153:Mutagens; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; D010365:Patulin; D053148:Caspase 3; D009955:Ornithine Decarboxylase