脑胶质瘤耐放射细胞亚株的建立方法

摘要

Objective To study the method of inducing and building glioma subline with radiation resistance ,and then to investigate the potential mechanism of the radioresistance in glioma cell.Methods The subject was human U251 glioma cell line, cultivating U251 cells in vitro, simulating the clinical radiotherapy , the U251 cells were repeatedly given individual dose with 60 Co resource to induce radiation resistance U 251 ( RR-U251 ) .After 5 Gy radiotherapy , the intracellular ultrastructure of the strains were observed by microscopy , MTT assay was examined the cell proliferation , the cell apoptosis rate was examined by flow cytometry ( FCM ) and the cell radiosensitivity were assayed by colony formation assay .Results The proliferation increased 11.5%, the apoptosis rate decreased 69.6%compared with the U251.Cell migration and invasion ability were increased 50%and 87.5%respectively .Cell clone analysis demonstrated that RR-U251 had greater plating efficiency (PE), after 5 Gy irradiation increased by 88.5%.Conclusion The method of radiating cells is viable to establish radioresistant glioma cell line-RR-U251.%目的：探索诱导并建立脑胶质瘤耐放射细胞亚株的体外实验方法，为进一步研究胶质瘤放疗抵抗发生的分子生物学机制提供基础。方法以脑胶质瘤U251细胞株为实验对象，通过体外细胞培养，应用60 Co射线对脑胶质瘤U251细胞株进行小剂量重复的诱导照射，得到U251耐放射细胞亚株RR-U251。观察RR-U251细胞形态的变化；MTT检测其细胞活力，流式细胞仪（ FCM）检测其细胞凋亡；集落形成实验检测细胞克隆团形成。结果放射治疗后，RR-U251与U251细胞的相比：细胞增殖活力增强11．5％，迁移能力增强50％，侵袭能力增强87．5％，凋亡率下降69．6％，细胞集落形成率升高88．5％；RR-U251放射敏感性明显下降。结论小剂量重复照射诱导法能较好地建立胶质瘤耐放射细胞亚株 RR-U251。