| 任精华,林菊生,刘瑶,等.γ-H2AX分析用于检测电离辐射致HepG2细胞DNA双链断裂的研究[J].中华放射医学与防护杂志,2007,27(5):450-454.RENJing-hua,LIN Ju-sheng,LIU Yao,et al.γ-H2AX assay as a measurement of DNA double-strand breaks in irradiated HepG2 cells[J].Chin J Radiol Med Prot,2007,27(5):450-454 |
| γ-H2AX分析用于检测电离辐射致HepG2细胞DNA双链断裂的研究 |
| γ-H2AX assay as a measurement of DNA double-strand breaks in irradiated HepG2 cells |
| 投稿时间：2006-10-19 |
| DOI： |
| 中文关键词: γ-H2AX DNA 双链断裂 电离辐射 |
| 英文关键词:γ-H2AX DNA double-strand breaks Ionizing radiation |
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| 中文摘要: |
| 目的 应用γ-H2AX 分析检测电离辐射导致肝癌细胞株HepG2 基因组DNA 双链断裂的情况, 并检测在不同细胞周期时相中的表达差异, 从而了解不同时相HepG2 细胞株的辐射敏感性。方法 利用胸腺核苷(TdR)阻断HepG2 细胞于G1期末, 于不同时间点分别得到同步化的S期和G2M 期细胞, 用60Co γ射线照射细胞, 建立DNA 双链断裂模型, 采用免疫荧光法和Westernblotting 检测γ-H2AX 的表达。结果 TdR 阻断后继续培养至34 和40 h, 分别得到了较高同步化程度的S 期和G2 M期HepG2 细胞; 受照后的HepG2 细胞中γ-H2AX 表达较照射前显著增高, 处于S 期的细胞γ-H2AX 表达增加更为突出。结论 不同周期时相的HepG2 细胞受照后均检测出不同程度的DNA 双链断裂, 其中S 期细胞尤为敏感。γ-H2AX 对DNA 双链断裂快速敏感的反应使γ- H2AX 分析在检测早期DNA 双链损伤中具有广泛的应用前景。 |
| 英文摘要: |
| Objective To explore the possibility of detecting DNA double-strand breaks (DSBs) in irradiated HepG2 cells by γ-H2AX assay and to investigate the γ-H2AX expression in different cell cycle phases. Methods HepG2 cells were blocked by TdR in G1 phase of cell cycle,and then cultured for different hours after TdR was removed. Synchronous S phase cells and G2/M phase cells were collected respectively and irradiated with 60Co γrays. Western blotting and immunofluorescence were used to analyze the expression ofγ-H2AX in cells. Results Synchronous S phase cells and G2/M phase cells were obtained at hours 34 and 40 post TdR block respectively. γ-H2AX level increased rapidly after cells were exposed to 60Co γ rays, especially in S phase of the cell cycle. Conclusions The cells in S phase are more prone to be damaged by ionizing radiation than those in G2/M.γ-H2AX assay can be regarded as an early diagnosis indicator of DSBs. |
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