| 樊婵,王豫,刘晓丹,黄波,徐勤枝,周平坤.细胞共培养旁效应实验模型的建立及其在低剂量α粒子诱发旁效应DNA损伤的应用[J].中华放射医学与防护杂志,2013,33(3):248-251 |
| 细胞共培养旁效应实验模型的建立及其在低剂量α粒子诱发旁效应DNA损伤的应用 |
| A cell co-culture model for studying bystander effect and its application on bystander DNA double-strand breaks induced by alpha-particles irradiation |
| 投稿时间：2013-01-08 |
| DOI：10.3760/cma.j.issn.0254-5098.2013.03.008 |
| 中文关键词: DNA双链断裂 旁效应 α粒子 免疫荧光染色 激光共聚焦 γH2AX簇集点 |
| 英文关键词:Double-strand breaks Bystander effect α-particle Immunofluorescence staining Laser confocal γH2AX |
| 基金项目:国家自然科学基金(81272994) |
|
| 摘要点击次数: 3821 |
| 全文下载次数: 2295 |
| 中文摘要: |
| 目的 建立研究旁效应DNA损伤的新的实验模型,探究低剂量α粒子辐射产生的旁效应DNA双链断裂(DSB)和修复的效应特征。方法 利用发红色荧光的HsBrk1-RFP融合蛋白标记其中一个细胞系,达到在共培养的条件下将直接受照细胞和非照射旁细胞区分开来,建立旁效应实验模型,α粒子照射细胞,γH2AX免疫荧光杂交和激光共聚焦显微观察检测DNA DSB损伤和修复动力学。结果 建立了HFS-RFP细胞和HFS细胞共培养的旁效应研究实验模型。无论是0.1 Gy还是0.2 Gy α粒子照射,直接受照细胞HFS-RFP和旁效应细胞HFS中的γH2AX簇集点数随时间变化的趋势相似,均在照后1 h达到高峰,随即减少,但在照射后8 h又出现一个高峰值,显示第二次DNA DSB损伤。旁效应二次DSB损伤要明显弱于直接受照细胞的二次损伤。结论 建立的细胞共培养旁效应实验模型能够成功地应用于低剂量的α粒子诱导的旁效应研究,并且低剂量的α粒子照射产生旁效应DSB损伤与修复的变化趋势与直接照射细胞的损伤相似,旁效应二次DSB损伤弱于直接照射细胞的二次损伤,可能与DNA簇集复杂损伤发生比例有关。 |
| 英文摘要: |
| Objective To establish an experimental model for the study of α-particle-induced bystander effect of DNA damage and investigate the characteristics of bystander DNA double-strand break (DSB).Methods The red fluorescence fusion protein of HsBrk1-RFP was used to mark the cytoplasm of one cell line to distinguish the irradiated target cells (HFS-RFP) and the non-irradiated bystander cells (HFS) in the co-culture cellular model. After α-particle irradiation, cellular DSB and its repair kinetics were analyzed by the immunofluorescence staining of γH2AX and laser confocal microscope observation. Results A bystander studying model was established by co-culturing human HFS-RFP cells with its partner HSF cells. After 0.1 Gy or 0.2 Gy α-particle irradiation, the similar kinetics of γH2AX foci production and abatement were observed in both irradiated HFS-RFP cells and non-irradiated bystander HFS cells, in which the highest level of γH2AX foci was detected at 1 h post-irradiation. The second peak of γH2AX foci formation appeared at 8 h post-irradiation, which possibly indicates the occurrence of secondary DSB. However, the production of secondary DSB in the bystander cells was weaker than that in the irradiated cells. Conclusions The cell co-culture model can be used for bystander effect investigation. Bystander DSB can be effectively induce by irradiation and the secondary breakage of DNA DSB in the bystander cells may relative to the consequential biochemical processing of clustered DNA damage. |
| HTML 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|