γH2AX对胶质瘤放射敏感性预测效果的实验研究  

Predictive effect of γH_2AX expression on the radiosensitivity of glioma

作　　者：王均伟[1] 张春智[2] 韩宝林[3] 王雷波[2] 姚鑫[2] 

机构地区：[1]天津医科大学研究生院2011级,300070 [2]天津市环湖医院神经外科,300060 [3]天津市宝坻区人民医院放疗科,301800

出　　处：《中国现代神经疾病杂志》2014年第3期240-244,共5页Chinese Journal of Contemporary Neurology and Neurosurgery

基　　金：天津市自然科学基金资助项目(项目编号:13JCYBJC21700);天津市卫生局科技公关项目(项目编号:12KG113);天津市卫生局科技基金资助项目(项目编号:2012K2003)~~

摘　　要：目的观察高级别胶质瘤细胞系(U87、U251和LN229)γH2AX蛋白表达变化,据此判断胶质瘤对放射线的敏感程度。方法选择胶质瘤细胞系U87、U251和LN229细胞株,通过细胞克隆形成实验,检测经不同剂量(0、2、4、6、8和10 Gy)X线照射后胶质瘤细胞克隆形成率,并绘制细胞存活曲线、测定放射敏感性;采用Western blotting法检测经剂量为2 Gy的X线照射后不同时间点(0 min、30 min、1 h、2 h、6 h、12 h、24 h、36 h和48 h)各胶质瘤细胞系γH2AX蛋白表达变化。结果细胞克隆形成实验显示,随着X线照射剂量的增加,胶质瘤细胞存活分数逐渐降低、细胞克隆形成率减少,放射增敏比自高至低依次为U87、LN229和U251细胞(均P=0.000)。Western blotting法显示,随着X线照射时间的延长,各胶质瘤细胞系γH2AX蛋白表达呈现先上升后下降的时间效应曲线,U87、LN229和U251细胞γH2AX蛋白表达峰值时间依次为2、1和1 h(P=0.000、0.000、0.015);γH2AX蛋白相对衰减速度(r=0.733,P=0.025)和升高程度(r=0.672,P=0.047)均与放射增敏比呈正相关。结论γH2AX蛋白有望成为检测高级别胶质瘤细胞系放射敏感性的一项预测指标。Objective To observe the expression changes of 3＇H2AX in high-grade glioma cell lines （U87, U251 and LN229） and to investigate the relationship between the expression of 3＇ H2AX and the radiosensitivity of high-grade glioma cells in vitro. Methods The radiosensitivity of glioma U251, U87 and LN229 cell lines were measured by clone forming assay. Afte X-ray irradiation of different doses （0, 2, 4, 6, 8 and 10 Gy）, the clone forming rates of 3 cell lines were measured, and cell survival curves were drawn. The DNA double-strand break （DSB） damage of 3 cell lines were determined by Western blotting assay. Results For glioma U251, U87 and LN229 cell lines, the survival fraction and clone forming rate were gradually decreased with the increase of X-ray radiation dose, and the radiotherapy sensitization enhancement ratio （SER） of U87 ceils was slightly higher compared with LN229, U251 cells （P = 0.000, for all）. In the Western blotting assay, the kinetics of the expression of γH2AX protein after irradiation was featured by increase and decay. The γH2AX expression of U87, LN229 and U251 cells after irradiation reached the peak value at 2 h, 1 h and 1 h respectively （P = 0.000, 0.000, 0.015）. There was positive correlation between SER and γH2AX attenuation speed （r = 0.733, P = 0.025）, as well as between SER and degree of increasing （r = 0.672, P = 0.047）. Conclusions The phosphorylated histone γH2AX is expected to become a powerful tool to monitor DNA DSB and to predict the radiosensitivity in high-grade glioma radiotherapy.

关 键 词：神经胶质瘤 细胞系 肿瘤 辐射耐受性 DNA损伤 细胞 培养的 

分 类 号：R739.4[医药卫生—肿瘤]