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首页> 外文期刊>Radiotherapy and oncology: Journal of the European Society for Therapeutic Radiology and Oncology >DNA double strand break quantification in skin biopsies.
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DNA double strand break quantification in skin biopsies.

机译：皮肤活检中的DNA双链断裂定量。

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BACKGROUND AND PURPOSE: Following induction of double strand breaks the histone H2AX is rapidly phosphorylated at regions flanking the breaks resulting in nuclear gammaH2AX foci. The purpose of this study was to use this endogenous signalling system to quantify the in vivo response to radiation in normal tissue. PATIENTS AND METHODS: Skin biopsies were taken from prostate cancer patients undergoing radiotherapy with a curative intent. Biopsies were taken at locations corresponding to 5 different doses in the range below 1.1Gy per fraction. Biopsies were taken from patients 30min following the first fraction and then once again following the fraction given after the first weekend break in the treatment course. The DNA double strand breaks were visualised as gammaH2AX foci using immunohistochemistry. Images were acquired using a CCD-camera and a fluorescence microscope and the gammaH2AX foci were quantified using digital image analysis including the basic procedures of top-hat transformation, threshold setting and labelling. RESULTS: Repeated assessments of the biopsies showed a high reproducibility in quantifying the number of foci per DNA area of the nucleated cells in epidermis. The reproducibility was equally good for the two biopsy occasions. A linear dose response was observed for the epidermis in the dose region 0-1Gy. CONCLUSIONS: We have established a method to measure the relative amount of DNA double strand breaks by detecting gammaH2AX foci in patients exposed to radiotherapy. The method provides a tool to study induction and repair of DNA double strand breaks and has the potential to predict individual radiosensitivity.
机译：背景和目的：诱导双链断裂后，组蛋白H2AX在断裂侧翼的区域迅速磷酸化，形成核gammaH2AX焦点。这项研究的目的是使用这种内源性信号系统来量化正常组织中体内对辐射的反应。患者和方法：皮肤活检取自接受放射治疗并有治愈意图的前列腺癌患者。在对应于每份低于1.1Gy的5种不同剂量的位置进行活检。在第一个疗程结束后30分钟从患者身上取活检，然后在治疗过程中第一个周末休息后再次进行活检。使用免疫组织化学将DNA双链断裂可视化为γH2AX焦点。使用CCD相机和荧光显微镜采集图像，并使用数字图像分析（包括大礼帽转换，阈值设置和标记的基本步骤）对gammaH2AX焦点进行定量。结果：对活组织检查的反复评估表明，在量化表皮中有核细胞的每个DNA区域的病灶数量方面具有很高的可重复性。两次活检的重现性同样好。在0-1Gy的剂量范围内观察到表皮的线性剂量反应。结论：我们已经建立了一种方法，可以通过检测放射治疗患者的gammaH2AX病灶来测量DNA双链断裂的相对量。该方法提供了研究DNA双链断裂的诱导和修复的工具，并具有预测个体放射敏感性的潜力。

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