目的:探讨真核翻译起始因子4E结合蛋白1(4E-BP1)与人脑胶质瘤细胞耐药性的关系,为寻找克服胶质瘤获得性耐药的新靶点提供理论依据。方法:Western blotting检测SWOZ2及其获得性耐药细胞株SWOZ2-BCNU中4E-BP1及其磷酸化蛋白(p-4E-BP1)的表达。小干扰RNA下调4E-BP1表达后,细胞计数试剂盒8检测SWOZ2细胞对卡氮芥(BCNU)的耐药性。免疫荧光法检测4E-BP1及p-4E-BP1(T70)蛋白在2株细胞中的表达位置。结果:SWOZ2-BCNU细胞中4E-BP1及其磷酸化蛋白表达较SWOZ2细胞明显下降。下调4E-BP1表达能明显增加敏感细胞对BCNU的耐药性。SWOZ2-BCNU细胞p-4E-BP1(T70)蛋白在获得性耐药过程中出现了核转位。结论:4E-BP1作为翻译调控关键蛋白,可能参与了人脑胶质瘤获得性耐药的发生机制。
AIM: For finding new target to overcome acquired drug resistance of glioma,the relationship between eukaryotic translation initiation factor 4E-binding protein 1(4E-BP1) and acquired drug resistance to carmustine [1,3-bis(2-chloroethyl)-1-nitrosourea,BCNU] in human glioma cells was investigated.METHODS: Western blotting was used to analyze the expression levels of 4E-BP1 and phosphorylated 4E-BP1(p-4E-BP1) in SWOZ2 cells and BCNU-resistant SWOZ2(SWOZ2-BCNU) cells.The siRNA targeting to human 4E-BP1 mRNA was transfected into SWOZ2 cells.Cell counting kit-8 was used to detect the sensitivity of the cells to BCNU before and after transfection.The location of 4EBP1 and p-4E-BP1(T70) in SWOZ2 cells and SWOZ2-BCNU cells was observed by the method of immunofluorescence.RESULTS: The expression of 4E-BP1 in SWOZ2-BCNU cells was significantly lower than that in SWOZ2 cells.Down-regulation of 4E-BP1 expression in SWOZ2 cells decreased the sensitivity of the cells to BCNU.The expression of p-4E-BP1(T70) in the nucleus of SWOZ2-BCNU cells was significantly higher than that in the cytosol.CONCLUSION: 4E-BP1 is involved in human glioma acquired drug resistance as a translation key protein.