摘要
目的探讨表皮生长因子受体(EGFR)拮抗剂-吉非替尼对人脑胶质瘤细胞的增殖抑制作用,为胶质瘤干细胞靶向治疗研究作准备。方法荧光定量PCR检测人脑胶质瘤细胞U251、GBM、SHG44及神经干细胞、脑肿瘤干细胞中EG-FR基因的表达。选择不同浓度的吉非替尼作用于U251细胞48h,MTT法检测药物对细胞的抑制率;流式细胞仪分析吉非替尼作用U251细胞后细胞增殖周期和细胞凋亡比率的变化。结果U251和脑肿瘤干细胞中都高表达EGFR。吉非替尼对U251细胞有明显的抑制作用,并呈浓度依赖性,IC50为19.0μM。以12.7μM、19.0μM和25.3μM吉非替尼处理U251细胞,随着药物浓度的增加,G0-G1期细胞比例和细胞凋亡率都逐渐升高。结论EGFR在U251细胞和脑肿瘤干细胞中都高表达,其拮抗剂吉非替尼对U251的细胞增殖有显著的抑制作用,为靶向治疗胶质瘤干细胞的研究开拓了思路。
Objective To investigate the inhibition effect of gefitinib, one of epidermal growth factor receptor （EGFR） inhibitors, on proliferation of human glioma cells in vitro, for the purpose of further studying the targeted therapy against glioma stem ceils. Methods The expression of EGFR was analyzed in glioma cell lines U251, GBM and SHG44, fetal neural stem cells and brain tumor stem cells with real-time PCR. After treatment with different concentration of gefitinib for 48 hours, the inhibition effect was assayed with MTF method, and the changes in tumor cell cycles and apoptosis were detected and quantified by flow cytometry. Results EGFR was highly expressed in U251 and brain tumor stem cells. After treatment with 0.5 to 128 μM of gefitinib for 48 hours, the growth of U251 cells were obviously inhibited in a positive dose-dependent manner, the 50% inhibitory concentration（ IC50 ） was 19.0μM. When U251 cells were treated with 12.7 μM, 19.0μM and 25.3 μM of gefitinib, respectively, the ratio of G0 - G1 phase tumor cells was increased from 64.43% to 82.66% compared with control group（50.15% ）, and the apoptosis ratio was increased from 13.01% to 40.36% compared with control group（ 1.45% ）. These results indicated that gefitinib inhibited U251 cells growth and promoted tumor cells apoptosis. Conclusions Overexpression of EGFR was found in U251 cells and brain tumor stem ceils. EGFR inhibitor, gefitinib, exerted significant inhibitory effort against proliferation of U251 cells in vitro, which deserve further study on targeted therapy for glioma stem ceils.
出处
《国际神经病学神经外科学杂志》
2009年第3期189-192,共4页
Journal of International Neurology and Neurosurgery
基金
国家自然科学基金(30672164)
