应用单细胞凝胶电泳技术检测放疗后椎体骨细胞DNA损伤

The determination of DNA damages of cell in vertebral body after radiotherapy by single cell gel electrophoresis

目的:建立检测放疗后椎体骨细胞DNA损伤的一种有效方法。方法:将20只比格犬以随机数字表法随机分为空白对照组(4只)与放射治疗组(16只)。在10～14 d内,对放疗组比格犬胸9～10椎体给予生物剂量为40～70 Gy的放疗。放射治疗结束3个月后,取空白及放疗组犬胸9～10节段椎体骨,在原提取骨细胞的方法上进行改良,首先用胰蛋白酶、胶原酶等消化骨块提取骨细胞,然后采用单细胞凝胶电泳技术检测放疗后椎体骨细胞DNA的损伤情况。结果:放疗能引起椎体骨细胞DNA断裂损伤,且损伤程度与放射剂量之间有明显的剂量-效应关系。结论:该技术简便、灵敏、准确、重现性好,可应用于放疗后椎体骨细胞DNA损伤的检测。

Objective： To establish a single cell gel electrophoresis method for the DNA damages of cell in vertebral body after radiotherapy. Methods. Select purebred beagles and 20 were divided into 2 groups randomly. Mimic a beagle clinical model of tumor in the ninth and tenth thoracic vertebrae. Then the second group beagles were irradiated by the radiotherapy, the total irradiation doses were 40-70 Gy in 10--14 d. The samples of vertebral body were taken out from the same locum of the nine and tenth thoracic vertebrae. On basis of the basic method of digested the bone cell, the bone of the vertebral body were pre-digested with trypsin and collagenase for harvesting osteoblasts. Then the DNA damages of cell in vertebral body were checked with single cell gel electrophoresis. Result： The DNA damages of cell in vertebral body can be caused by radiotherapy. The DNA damages of cell and the dose of the radiotherapy has a good dosage-domino effect. Conclusions： This method is convenient, sensitive and accurate. And it can be used to tested the DNA damages of cell in vertebral body after radiotherapy.