title: XPC silencing sensitizes glioma cells to arsenic trioxide via increased oxidative damage.
abstract: Arsenic exerts its cytotoxicity via the generation of reactive oxygen species and inhibition of DNA repair. How arsenic disturbs oxidative DNA damage repair is, however, unclear. We found that arsenic trioxide (ATO), like ultraviolet (UV) irradiation, induced the expression of xeroderma pigmentosum group C (XPC) but not of xeroderma pigmentosum A in a human glioma cell line, U87. To explore the role of XPC in the toxic effects of ATO, small interfering RNA was used to silence XPC (siXPC) in U87 cells. siXPC cells were more susceptible to UV irradiation and ATO-induced cell death than control cells. Increased siXPC cell death induced by ATO was accompanied by increased senescence and autophagy. Because increased DNA strand breaks in siXPC cells were observed only when cells were concomitantly treated with ATO and DNA repair inhibitors, XPC silencing apparently did not interfere with repair of ATO-induced DNA damage. Although intracellular ROS levels were not significantly enhanced in siXPC cells, ATO treatment did result in increased 8-hydroxy-2'-deoxyguanosine and hyperoxidized peroxiredoxin. Enhanced superoxide production and autophagy by ATO in siXPC cells were suppressed by co-incubation with N-acetylcysteine (NAC). Furthermore, XPC silencing caused decreased glutathione levels and increased catalase and Mn-superoxide dismutase activities. Increased catalase activity in siXPC cells was suppressed by ATO treatment. XPC silencing also enhanced reporter activity of activator protein-1, whereas enhanced activity was suppressed by NAC. Taken together, our results indicate that XPC silencing causes increased ATO susceptibility by disturbing redox homeostasis rather than reducing DNA repair.
authors.affiliation: Department of Biomedical Image and Radiological Sciences, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Shin-Yi
authors.initials: SY
authors.lastname: Liu
authors.forename: Ching-Ya
authors.initials: CY
authors.lastname: Wen
authors.forename: Yi-Jang
authors.initials: YJ
authors.lastname: Lee
authors.forename: Te-Chang
authors.initials: TC
authors.lastname: Lee
mesh_terms: D000077237:Arsenic Trioxide; D001152:Arsenicals; D001343:Autophagy; D001483:Base Sequence; D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D016922:Cellular Senescence; D004268:DNA-Binding Proteins / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D020868:Gene Silencing; D005910:Glioma / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D018384:Oxidative Stress; D010087:Oxides / Q000633:toxicity*; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D001152:Arsenicals; D004268:DNA-Binding Proteins; D010087:Oxides; D034741:RNA, Small Interfering; C086984:XPC protein, human; D000077237:Arsenic Trioxide