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.
出处
《生物技术通讯》
CAS
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)