title: Role of type-II pathway in apoptotic cell death induction by photosensitized CDRI-97/78 under ambient exposure of UV-B.
abstract: Novel trioxane 97/78, developed by Central Drug Research Institute (CDRI), Lucknow has shown promising antimalarial activity. Clinical experience of anti-malarial drugs registered the occurrence of phototoxicity in patients exposed with sunlight subsequent to medication. Photodegradation study has identified one photo-product up to 4h under UV-B/Sunlight by LC-MS/MS. UV-B irradiated 97/78 compound produced ¹O₂ via type-II dependent reaction mechanism, corroborated by its specific quencher. 2'-dGuO degradation and % tail development in photochemical as well as comet test, advocated the genotoxic potential of 97/78. The photocytotoxicity assays (MTT and NRU) on HaCaT cell line revealed the considerable decline in cell viability by 97/78. Cell cycle and Annexin V/PI double stain along with AO/EB demonstrated the G2/M phase arrest and apoptosis. Significant caspase-3 activity was measured in photoexcited 97/78 by colorimetric assay. Fluorescence stain with AO/JC-1 confirmed the lysosomal disruption and mitochondrial membrane destabilization by UV-B irradiated 97/78. Gene expression by RT-PCR showed significant upregulation of p21 and pro-apoptotic Bax, but no change observed in Bcl-2. In conclusion, the study highlights ROS mediated DNA damage, lysosomal and mitochondrial destabilization via upregulation of Bax and activation of caspase-3 which further leads to apoptosis.
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow, 226001, India.
authors.forename: Ashish
authors.initials: A
authors.lastname: Dwivedi
authors.forename: Manish Kumar
authors.initials: MK
authors.lastname: Pal
authors.forename: Amit Kumar
authors.initials: AK
authors.lastname: Tripathi
authors.forename: Neera
authors.initials: N
authors.lastname: Yadav
authors.forename: Syed Faiz
authors.initials: SF
authors.lastname: Mujtaba
authors.forename: M C
authors.initials: MC
authors.lastname: Pant
authors.forename: Shio Kumar
authors.initials: SK
authors.lastname: Singh
authors.forename: Durga Prasad
authors.initials: DP
authors.lastname: Mishra
authors.forename: Ratan Singh
authors.initials: RS
authors.lastname: Ray
authors.forename: B H
authors.initials: BH
authors.lastname: Manjunatha Prabhu
mesh_terms: D000962:Antimalarials / Q000009:adverse effects* / Q000737:chemistry / Q000528:radiation effects; D017209:Apoptosis / Q000187:drug effects*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000009:adverse effects* / Q000737:chemistry / Q000528:radiation effects; D053148:Caspase 3 / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004249:DNA Damage; D017484:Dermatitis, Phototoxic / Q000378:metabolism* / Q000473:pathology; D016195:G2 Phase / Q000187:drug effects; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008247:Lysosomes / Q000187:drug effects / Q000473:pathology; D051336:Mitochondrial Membranes / Q000187:drug effects / Q000473:pathology; D010782:Photolysis / Q000528:radiation effects; D017319:Photosensitizing Agents / Q000009:adverse effects* / Q000737:chemistry / Q000528:radiation effects; D016283:Proto-Oncogene Proteins p21(ras) / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D026082:Singlet Oxygen / Q000737:chemistry; D013472:Sunlight; D014466:Ultraviolet Rays; D015854:Up-Regulation / Q000187:drug effects; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
chemical_list: D000962:Antimalarials; C494826:BAX protein, human; D019086:Bridged Bicyclo Compounds, Heterocyclic; C574493:CDRI-97-78; D017319:Photosensitizing Agents; D051028:bcl-2-Associated X Protein; D026082:Singlet Oxygen; C505407:CASP3 protein, human; D053148:Caspase 3; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)
keywords: Antimalarial; DNA damage; LC–MS/MS; Mitochondrial membrane destabilization; UV-B