title: Benzophenone 1 induced photogenotoxicity and apoptosis via release of cytochrome c and Smac/DIABLO at environmental UV radiation.
abstract: Solar UV radiation is main factor of photocarcinogenesis, photoageing, and phototoxicity; thus, protection from UV radiation is major concern. Sunscreens containing UV filters are suggested as sun safe practices, but safety of UV filters remains in controversies. Benzophenone-1 (BP1) is commonly used in sunscreens as UV blocker. We assessed the photogenotoxicity and apoptotic parameters in human keratinocytes (HaCaT cells) by western blot, immunocytochemistry, flowcytometry, comet assay and TEM imaging. Our results exposed that BP1 photosensitized and generated intracellular ROS (2.02 folds) under sunlight/UVR. Decrease in cell viability was recorded as 80.06%, 60.98% and 56.24% under sunlight, UVA and UVB, respectively. Genotoxic potential of BP1 was confirmed through photomicronuclei and CPDs formation. BP1 enhanced lipid peroxidation and leakage of LDH enzyme (61.7%). Apoptotic cells were detected by AnnexinV/PI staining and sub G1 population of cell cycle. BP1 induced up regulation of apoptotic proteins Bax/Bcl2 ratio, Apaf-1, cytochrome c, Smac/DIABLO and cleaved caspase 3 was noticed. Down regulation of pro caspase 3 was inhibited by Z-VAD-fmk (inhibitor of caspase). Thus, study established the involvement of BP1 in photogenotoxicity and apoptosis via release of cytochrome c and Smac/DIABLO. These findings suggest sunscreen user to avoid BP1 in cosmetics preparation for its topical application.
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India; Academy of Scientific and Innovative Research, CSIR-IITR Campus, Lucknow, India.
authors.forename: Saroj Kumar
authors.initials: SK
authors.lastname: Amar
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India; Academy of Scientific and Innovative Research, CSIR-IITR Campus, Lucknow, India.
authors.forename: Shruti
authors.initials: S
authors.lastname: Goyal
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Divya
authors.initials: D
authors.lastname: Dubey
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Ajeet K
authors.initials: AK
authors.lastname: Srivastav
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Deepti
authors.initials: D
authors.lastname: Chopra
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Singh
authors.affiliation: Transmission Electron Microscopy Laboratory, CSIR-IITR, Lucknow, India.
authors.forename: Jai
authors.initials: J
authors.lastname: Shankar
authors.affiliation: Developmental Toxicology, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Rajnish K
authors.initials: RK
authors.lastname: Chaturvedi
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, P.O. Box 80, M.G. Marg, Lucknow 226001, India. Electronic address: ratanray.2011@rediffmail.com.
authors.forename: Ratan Singh
authors.initials: RS
authors.lastname: Ray
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins; D053477:Apoptotic Protease-Activating Factor 1 / Q000235:genetics / Q000378:metabolism; D001577:Benzophenones / Q000633:toxicity*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D020552:Comet Assay; D045304:Cytochromes c / Q000378:metabolism*; D004249:DNA Damage / Q000187:drug effects*; D015536:Down-Regulation; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial; D046529:Microscopy, Electron, Transmission; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013473:Sunscreening Agents / Q000737:chemistry; D014466:Ultraviolet Rays / Q000009:adverse effects*; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: C506592:APAF1 protein, human; D000590:Amino Acid Chloromethyl Ketones; D017304:Annexin A5; D051017:Apoptosis Regulatory Proteins; D053477:Apoptotic Protease-Activating Factor 1; C494826:BAX protein, human; D001577:Benzophenones; C412403:DIABLO protein, human; D047908:Intracellular Signaling Peptides and Proteins; D024101:Mitochondrial Proteins; D017382:Reactive Oxygen Species; D013473:Sunscreening Agents; D051028:bcl-2-Associated X Protein; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C047723:benzophenone; D045304:Cytochromes c; C505407:CASP3 protein, human; D053148:Caspase 3
keywords: Caspase 3; Cytochrome c; DNA damage; Photomicronuclei; Phototoxicity; Smac/DIABLO