阿帕替尼对脑胶质瘤细胞放射敏感性的影响

Effect of apatinib on radiosensitivity of glioma cells

目的 研究小分子酪氨酸激酶抑制剂甲磺酸阿帕替尼对脑胶质瘤细胞U87MG放射敏感性的影响,并初步探讨其作用机制.方法 将胶质瘤细胞U87MG分为空白对照组、阿帕替尼组、单纯照射组、阿帕替尼联合照射组.CCK-8法检测不同浓度阿帕替尼(5、10、20、40、80 μmol/L)对细胞增殖的影响;创伤愈合实验和侵袭实验检测阿帕替尼对胶质瘤细胞的迁移及侵袭能力的影响;平板克隆实验检测阿帕替尼对脑胶质瘤细胞的放射敏感性的影响;流式细胞术检测细胞凋亡情况;采用Western blot检测Bax、Bcl-2蛋白表达.结果 阿帕替尼对胶质瘤细胞U87MG的增殖具有明显抑制作用,且存在时间-剂量的依赖性.与单纯照射组比较,阿帕替尼联合照射组能明显抑制细胞的增殖、迁移和侵袭,差异有统计学意义(t=9.857、18.704、4.167,P< 0.05).与单纯照射组比较,阿帕替尼联合照射组的D_(0)、D_(q)和SF_(2)均下降,放射增敏比(SER_(D0))为1.3.与单纯照射组比较,阿帕替尼联合照射组的细胞凋亡率增加,Bcl-2蛋白表达降低,Bax蛋白表达升高,差异有统计学意义(t=16.187、8.890、5.222,P<0.05).结论 阿帕替尼能够抑制胶质瘤细胞增殖、迁移和侵袭,并促进细胞凋亡,增加胶质瘤细胞的放射敏感性.

Objective To investigate the effect of apatinib on radiosensitivity of glioma cells U87MG and its potential mechanism.Methods U87MG cells were divided into control group,apatinib group,radiation group and combination group treated with apatinib and radiation.The effect of different concentrations of apatinib(5,10,20,40,80μmol/L)on cell proliferation was detected by CCK8 assay.The effect of apatinib on cell migration and invasion was detected by wound-healing assay and transwell assay,respectively.The effect of apatinib on cell radiosensitivity was detected by plate cloning assay,the cell apoptosis rate was detected by flow cytometry,and the protein expressions of Bax and Bcl-2 were detected by Western blot.Results Apatinib significantly inhibited the proliferation of U87MG cells in a manner depended on the drug treatment time and radiation.Compared with the radiation group,the cell proliferation,migration and invasion in the combination group were inhibited much significantly(t=9.857,18.704,4.197,P<0.05),so that the value of D_(0),D_(q) and SF_(2) of the combination group was lower,resulting in a radiosensitivity enhancement ratio(SER_(D0))of 1.3.Moreover,compared with the radiation group,the apoptosis rate of the combination group was increased,the expression of Bcl-2 protein was decreased,and the expression of Bax protein was increased(t=16.187,8.890,5.222,P<0.05).Conclusions Apatinib inhibits cell proliferation,invasion and migration,induces apoptosis and increases radiosensitivity of glioma cells.