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目的探讨化疗耐药与肿瘤干细胞之间的关系。方法分析替莫唑胺(TMZ)作用前后胶质瘤U251中CD133蛋白表达及阳性细胞数量的变化。通过MTT实验评估U251细胞对TMZ的敏感性。利用免疫荧光染色和Western blot检测TMZ作用前后U251细胞中CD133、巢蛋白(nestin)、神经元特异性烯醇化酶(NSE)、神经胶质细胞纤维酸性蛋白(GFAP)等蛋白表达。结果U251细胞中未加入TMZ组的CD133+细胞比例约为0.060±0.003,明显低于加入TMZ组的0.337±0.012(P<0.05)。在TMZ作用后,CD133和nestin表达明显增加,而GFAP和NSE表达明显减少(P<0.05)。结论胶质瘤U251细胞株中存在CD133+细胞,大部分具有脑肿瘤干细胞特性;在TMZ作用后,这类细胞比例增加,提示其与化疗耐药有关。
Objective To investigate the relationship between chemoresistance and tumor stem cells. Methods The expression of CD133 and the number of positive cells in U251 glioma before and after temozolomide (TMZ) treatment were analyzed. The sensitivity of U251 cells to TMZ was assessed by MTT assay. The expression of CD133, nestin, neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) in U251 cells before and after TMZ treatment were detected by immunofluorescence staining and Western blot. Results The proportion of CD133 + cells in U251 cells without TMZ group was about 0.060 ± 0.003, which was significantly lower than that in TMZ group (0.337 ± 0.012, P <0.05). After TMZ treatment, the expression of CD133 and nestin increased significantly, while the expression of GFAP and NSE decreased significantly (P <0.05). Conclusion CD133 + cells exist in glioma U251 cell line, most of which have the characteristics of brain tumor stem cells. After TMZ treatment, the proportion of these cells increases, suggesting that they are associated with chemoresistance.