CAJ | 학술논문

目的:用建立的稳定表达脆性组氨酸三联体(Fhit)突变体的细胞株,研究 Fhit 与复制蛋白 A(RPA)之间的相互作用对细胞在 DNA 损伤后的影响.方法:用电离辐射或 DNA 损伤诱导剂喜树碱处理稳定表达 Fhit 突变体的阳性细胞株 HeLa-FhitA/D/F 后,通过流式细胞技术、MTT 比色法及克隆形成实验,检测这些细胞系的细胞周期变化情况以及对 DNA 损伤诱导剂的敏感性.结果:DNA 损伤诱导剂处理后,Fhit 突变体基的高表达可以使细胞表现出更强的G2期阻滞及对 DNA 损伤诱导剂更耐受.结论:Fhit 与 RPA 相互作用的改变影响了细胞对 DNA 损伤诱导剂的耐受性,为阐明 Fhit 在维持基组完整性方面的机理提供了线索.
목적:용건립적은정표체취성조안산삼련체(Fhit)돌변체적세포주,연구 Fhit 여복제단백 A(RPA)지간적상호작용대세포재 DNA 손상후적영향.방법:용전리복사혹 DNA 손상유도제희수감처리은정표체 Fhit 돌변체적양성세포주 HeLa-FhitA/D/F 후,통과류식세포기술、MTT 비색법급극륭형성실험,검측저사세포계적세포주기변화정황이급대 DNA 손상유도제적민감성.결과:DNA 손상유도제처리후,Fhit 돌변체기적고표체가이사세포표현출경강적G2기조체급대 DNA 손상유도제경내수.결론:Fhit 여 RPA 상호작용적개변영향료세포대 DNA 손상유도제적내수성,위천명 Fhit 재유지기조완정성방면적궤리제공료선색.
Objective: To explore the role of fragile histidine triad(Fhit) and replication protein A(RPA) interac?tion in mammalian cells to DNA damage response by using established stable expressed cell lines of human Fhit mutants. Methods: After treatment with ionizing radiation(IR) or camptothecin(CPT), the stable cell lines HeLa-FhitA/D/F expressing human Fhit mutants were harvested at different time and cell cycles and cell survival fractions were tested by using flow cytometry or MTT and colony-forming assay. Results: The effect of Fhit mu?tant in mammalian cells to the cell cycles and the sensitivities to IR or CTP-induced DNA damage was identi?fied, it was found that these cells showed stronger G2 checkpoint response to IR and more resistant to CPT-in?duced killing than Fhit wild type cells. Conclusion: The interaction between Fhit and RPA affects the radiosensi?tivity of cells to DNA damage inducer, it provides a clue to elucidate the mechanism for Fhit maintaining genom?ic integrity.