肿瘤睾丸抗原CT45-5在DNA损伤应答中的作用  

Role of Cancer Testis Antigen CT45-5 in DNA Damage Response

作　　者：柯波[1,2] 绳纪坡[1] 于芳[1] 林宇翔[1,2] 罗维[1,2] 胡宝成[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]福建农林大学生命科学院,福建福州350002

出　　处：《生物技术通讯》2013年第2期157-160,共4页Letters in Biotechnology

基　　金：国家自然科学基金(31070760;30770651;30670616)

摘　　要：目的:用建立的CT45-5-siRNA研究肿瘤睾丸抗原CT45-5在DNA损伤应答中的作用。方法:通过靶向CT45-5基因小干扰RNA(siRNA)下调CT45-5的表达,用MTT比色法检测经喜树碱和依托泊苷处理后细胞对DNA损伤诱导剂的敏感性,用流式细胞技术检测电离辐射后细胞周期的变化。结果:在DNA损伤诱导剂处理后,在脆性组氨酸三联体(Fhit)高表达细胞中下调CT45-5的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,而在Fhit缺失表达的细胞中却没有此现象。结论:CT45-5可能是以Fhit依赖的途径参与了DNA损伤应答。Objective： To investigate the role of cancer testis antigen CT45-5 in DNA damage response by using CT45-5-siRNA. Methods： After CT45-5 gene was knocked down by siRNA and treatment with camptothecin （CPT） or etoposide（Et） or ionizing radiation（IR）, the stable cell lines 3-18, overexpressing human fragile histi- dine triad（Fhit）, were harvested at different time, cell survival fractions and cell cycles were tested by using MTT assay or flow cytometry. Results： The cells overxpressing Fhit by knocking down CT45-5 gene showed stronger G2 checkpoint response to IR and more resistant to CPT and Et-induced killing than control cells following DNA damage inducer. Conclusion： CT45-5 may involve in the DNA demage response by Fhit-dependent pathway.

关 键 词：肿瘤睾丸抗原CT45—5 RNA干扰 DNA损伤应答 脆性组氨酸三联体 

分 类 号：Q25[生物学—细胞生物学] Q78