| 罗加林,曹建平,朱 远.胞质分裂阻滞微核法研究ATM基因辐射防护功能[J].肿瘤学杂志,2013,19(5):390-394. |
| 胞质分裂阻滞微核法研究ATM基因辐射防护功能 |
| A Study on Radiation Protective Function of ATM Gene Using Cytokinesis-block Micronucleus Method |
| 投稿时间：2013-03-15 |
| DOI：10.11735/j.issn.1671-170X.2013.05.B201303110 |
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| 中文关键词: 毛细血管扩张性共济失调症 电离辐射 微核 辐射耐受性 |
| 英文关键词:ataxia-telangiectasia ionizing radiation micronucleus radiation tolerance |
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| 中文摘要: |
| 摘 要：［目的］ 研究ATM基因对毛细血管扩张性共济失调症(AT)患者皮肤成纤维细胞系AT5BIVA（AT细胞）高辐射敏感性的纠正情况，以评价ATM基因的辐射防护功能。［方法］ 利用电穿孔技术，将含有ATM基因cDNA 的真核表达载体PEBS7-YZ5转染到AT细胞，用潮霉素筛选以获得稳定表达细胞株，RT-PCR检测ATM cDNA的转录，Western blot验证ATM蛋白的表达；用胞质分裂阻滞微核法（CBMN），在PEBS7-YZ5-AT细胞、PEBS7-AT细胞和未转染AT细胞经60CO γ射线0、1、2、3、4Gy照射后，观察比较三者间微核率及微核细胞率的差异。［结果］ PEBS7-YZ5成功转进AT细胞，RT-PCR检测到ATM cDNA片段，Western blot检测到ATM蛋白的表达。在0、1、2、3、4Gy剂量下，PEBS7-YZ5-AT细胞微核率及微核细胞率明显低于PEBS7-AT细胞和AT细胞，其差异具有显著统计学意义(P＜0.01）；而PEBS7-AT细胞和AT细胞间无明显差异(P＞0.05）。［结论］ AT细胞高辐射敏感性被ATM基因纠正，ATM基因具有辐射防护功能，含有ATM基因的真核表达载体PEBS7-YZ5有望成为辐射防护剂。 |
| 英文摘要: |
| Abstract:［Purpose］ To investigate the role of ATM gene in rectifing the high radiosensitivity of fibroblast AT5BIVA cell line which established from the skin of patients with ataxia-telangiectasia (AT)，and to evaluate the radiation protection function. ［Methods］ PEBS7-YZ5 containing ATM gene cDNA was transfected into AT cells by electroporation. Hygromicin was used to screen the cells with ATM protein stably expressed. RT-PCR was used to detect the expression level of ATM. Western blot was used to detect the expression of ATM protein. Using cytokinesis-block micronucleus method，the difference of micronucleus frequencies(MNF) and micronucleus cell frequencies(MNCF) of PEBS7-YZ5-AT cells，PEBS7-AT cells and AT cells treated with 0，1，2，3 and 4Gy exposures to 60CO γ-ray were observed.［Results］ PEBS7-YZ5 was transfected into AT cells successfully. Fragment of ATM cDNA and ATM protein were detected by RT-PCR and Western blot respectively. After exposed to 1，2，3 and 4Gy 60CO γ-ray，the radiation-induced level of MNF and MNCF were significantly lower in the PEBS7-YZ5-AT cells，compared with those in PEBS7-AT cells and AT cells(P＜0.01），and there was no significant difference between PEBS7-AT cells and AT cells(P＞0.05）. ［Conclusion］ ATM gene rectifies the high radiosensitivity of AT cells. ATM gene pocesses the function of radiaiton protection，and PEBS7-YZ5 plasmid is potential be radiation protection agent. |
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