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选择性COX-2抑制剂对放射所致DNA损伤修复的影响

Effects of Selective COX-2 Inhibitor on DNA Repairing after Irradiation

  • 摘要: 目的选择性COX-2抑制剂celecoxib对放射引起的DNA损伤修复的影响。方法流式细胞仪Sub-G1法检测凋亡率和细胞周期;单细胞凝胶电泳技术检测DNA的损伤及修复。RT-PCR和Western blot检测DNA修复基因ku70、ku80的表达。结果celecoxib对HeLa细胞的周期分布无明显改变,但可引起剂量依赖性凋亡。celecoxib可明显提高放射引起的凋亡率,剂量增强比(DEF)在60、80、100μmol/L时分别为3.18、4.16、6.96。同时celecoxib对6Gy X线引起的G2/M期阻滞有明显的减弱作用(放射组:G2/M32.35%,药放组:G2/M5.95%);celecoxib不明显增加放射引起的DNA损伤,但可明显延缓放射引起的SSB和DSB的修复(P<0.05);celecoxib明显抑制HeLa细胞ku70、ku80的表达(P<0.05)。结论COX-2选择性抑制剂celecoxib对肿瘤细胞的放射增敏机制可能与抑制DNA损伤修复有关,其作用可能通过减弱辐射诱导的G2/M期阻滞和抑制DSB重组修复酶DNA-PK成分ku的表达得以实现。

     

    Abstract: Objective  To investigate the effect s of a selective COX-2 inhibitor-celecoxib on DNA repairing af ter X-rays irradiation, in order to elucidate the possible mechanism of the radiosensitization of selective COX-2 inhibitor. Methods  Human cervical carcinoma cells ( HeLa) were cultured in vit ro and exposed to different doses of celecoxib for 48h then with and without radiation ( 6 Gy), flow cytomet ry sub-G1fluorescence parameter line was performed to analyze the change of the apoptosis and cell cycle dist ri-bution. The DNA damage and repair were detected by single cell gel elect rophoresis assay. Semi-quantita-tive RT2PCR and Western blot were used to measure the mRNA and protein level of ku70 and ku80 in HeLa cell lines t reated with celecoxib. Results  Celecoxib had no effect on the cell cycle dist ribution of HeLa cell lines, but it induced apoptosis increasing dependent on celecocib doses. Radiation-induced apop-tosis was enhanced af ter Hela cells t reated with celecoxib (enhancement factor = 6. 96 at100μmol/ L), and the radiation2induced G2/ M arrest was markedly decreased by celecoxib〔Radiation group ( RT) : G2 / M 32. 35 %, 100μmol/ L celecoxib + RT : G2 / M 5. 95 %〕. The DNA damage induced by radiation was not increased, but the DNA repair was reduced in Hela cells t reated with celecoxib (60μmol/ L) for 48h, the mRNA and protein level of ku70 and ku80 were also significantly decreased ( P < 0. 05) . Conclusion  The result s suggest that the possible mechanism of radiosensitization of a selective COX-2 inhibitor was relat-ed to inhibition of DNA repair including reduced G2/ M arrest cells and decreased DNA-PK expression.

     

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