【摘要】 目的探讨埃罗替尼(Erlotinib)和雷帕霉素(Rapamycin)抑制胶质瘤细胞增殖的可能机制,为胶质瘤靶向治疗提供新思路。方法采用无血清培养基培养C6细胞,获取C6胶质瘤干细胞(GSCs),细胞免疫荧光染色检测CD133和nestin鉴定胶质瘤干细胞;CCK-8、RT-PCR和Western blot检测经Erlotinib、Rapamycin干预后,肿瘤细胞增殖和细胞周期与EGFR和mTOR信号通路相关基因蛋白EGFR、Akt、p-Akt、mTOR、p-mTOR、rps6k、p-rps6k表达变化的关系。结果胶质瘤干细胞表达CD133、Nestin;Erlotinib、Rapamycin对C6及C6 GSCs均呈剂量-时间依赖性抗增殖作用(P<0.05)。Erlotinib与Rapamycin能够增加Gl期细胞比例,减少S期比例,而且C6 GSCs对Erlotinib、Rapamycin的敏感性低于C6细胞(P<0.05);用药组与空白对照组相比,细胞中EGFR和mTOR通路中相关蛋白表达明显下调(P<0.05),联合用药组效果更明显。结论胶质瘤C6细胞系中存在一定量的肿瘤干细胞;Erlotinib与Rapamycin通过阻断EGFR和mTOR抑制C6、C6 GSCs生长增殖、细胞周期。更多还原

【Abstract】 Objective To investigate the mechanism of inhibiting glioma cells proliferation with Erlotinib and Rapamycin,and provide new methods for glioma targeted therapy.Methods C6 cells were cultured by the serum free medium,and the tumor stem cells(GSCs) were obtained.Cell immunofluorescence staining was used for detection of CD133 and nestin to identify the tumor stem cells;CCK-8,RT-PCR and Western blot were used to observe the relationship of tumor cell proliferation,cell cycle and EGFR,mTOR signal pathways related genes,and the expressions of proteins(EGFR,Akt,p-Akt,mTOR,p-mTOR,rps6k,p-rps6k) after intervention of Erlotinib and Rapamycin.Results The expressions of CD133 and nestin were detected in the glioma stem cells;Erlotinib and Rapamycin had the dose-time dependence antiproliferative effect in C6 and C6 GSCs(P<0.05).Erlotinib and Rapamycin can increase the ratio of cells in G1 phase and reduce the ratio of cells in S phase.The sensitivity to Erlotinib and Rapamycin in GSCs was lower than that in C6 cells(P<0.05);protein expressions of EGFR and mTOR pathways in the treatment group were lower than those in the control group(P<0.05),the efficacy is significant in the combined treatment group.Conclusion Tumor stem cells exist in C6 glioma cell lines.Erlotinib and Rapamycin can inhibit the proliferation and cell cycle of C6 and C6 GSCs by means of blocking the EGFR and mTOR signaling pathways.更多还原