γ射线致旁效应细胞脆性组氨酸三联体的表达  

Expression of FHIT in AHH-1 cells irradiated by ^60 Co γ-ray and bystander effect cells

作　　者：高娴[1] 孙顶[1] 杨剑[1] 秦阳华[1] 肖瑶[1] 韩玲[1] 

机构地区：[1]第二军医大学放射医学教研室,上海200433

出　　处：《中华放射医学与防护杂志》2008年第3期217-220,共4页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金资助项目（10575130）

摘　　要：目的研究^60Coγ射线照射后人淋巴母细胞（AHH-1）及其旁效应细胞中脆性组氨酸三联体（FHIT）基因的表达情况，探讨FHIT基因在电离辐射旁效应中的作用。方法旁效应细胞模型的制备：分别用不同剂量（0、2和5Gy）^60Coγ射线照射AHH-1细胞后，立即离心，将直接受照细胞与未受照细胞共同培养于Transwell中，此未受照细胞为旁效应1组；另将直接受照细胞培养液转移至未受照细胞培养瓶中形成旁效应2组。于照后0、6、12和24h收集细胞，抽提细胞总RNA及总蛋白，分别应用RT-PCR和Westernblot方法检测FHITmRNA及蛋白表达水平；并在照后即刻和24h收集细胞，应用流式细胞仪检测细胞周期，并应用细胞计数法观察细胞增殖变化。结果对照细胞、直接受照细胞、旁效应细胞中FHIT基因mRNA水平变化不明显，而FHIT蛋白表达在直接受照细胞及旁效应细胞中显著下调（F=102．45．P〈0．001）。细胞周期分析结果显示直接受照细胞及旁效应细胞G2期阻滞明显，且增殖能力明显下降。结论2、5Gy^60Coγ射线照射能导致直接照射AHH-1细胞及其旁效应细胞FHIT蛋白表达明显下调，提示FHIT基因在电离辐射旁效应中发挥一定作用。Objective To investigate the expression of FHIT gene in the ^60 Co gamma-ray irradiated human lymphocytoblast （AHH-1） cell and the bystander effect cell, and to explore the function of FHIT gene in the bystander effect of ionizing radiation. Method Preparation of bystander effect cell model： after irradiated with different dose of ^60 Co gamma-ray （0, 2, 5 Gy）, the directly irradiated AHH-1 cells were collected immediately by eentrifugation and co-cultivated with non-irradiated cells in Transwell, forming the bystander effect group P1. In addition, some culture media supematant of directly irradiated cells were transferred to the non- irradiated cells culture medium, forming the group P2. Then cells were collected at 0, 6, 12, and 24 h after irradiation and the total RNA and protein were extracted. RT-PCR and Western blot were performed to determine the FHIT mRNA and protein level, respectively. Flow eytometry assay and cell counting were conducted to detect the alteration of cell cycle and cell proliferation, respectively at 0, 24 h after irradiation. Results The mRNA level of FHIT gene among control cells, directly irradiated cells and bystander cells showed no obvious difference, while the FHIT protein level of the directly irradiated cells and bystander cells was significantly down-regulated compared with the control cells （ F = 102.45, P 〈 0.001 ）. Moreover, the directly irradiated cells and bystander cells showed significant G2 phase arrest and obviously inhibited the proliferation ability. Conclusions 2 and 5 Gy of ^60Co γ-ray irradiated AHH-1 cells can result in down regulation of the FHIT protein expression, which suggests that FHIT gene is involved in the process of bystander effect induced by irradiation.

关 键 词：^60CO Γ射线 旁效应 AHH-1细胞 FHIT 

分 类 号：R363[医药卫生—病理学]