干扰素抗MGMT阳性胶质瘤干细胞替莫唑胺的耐药机制  

Mechanism by which interferon reduces the resistance of MGMT positive glioma stem cells to temozolomide

作　　者：苏辉 刘兆玮 杜红利 乔艳梅 

机构地区：[1]衡水市人民医院,河北省衡水市053000

出　　处：《中国组织工程研究》2015年第36期5800-5805,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:临床对胶质瘤患者进行治疗的过程中容易出现耐药现象,研究发现,MGMT是胶质瘤表达的一种多药耐药基因。目的:探讨干扰素增加替莫唑胺对MGMT阳性胶质瘤干细胞的增敏作用。方法:采用"悬浮克隆球形成法"对胶质瘤细胞株U251、SKMG-4进行诱导,获得MGMT阳性的胶质瘤干细胞U251G、SKMG-4G。应用CCK-8法检测干扰素α/β联合替莫唑胺对MGMT阳性胶质瘤干细胞的杀伤效应;RT-PCR和Western blot检测预用干扰素α/β后MGMT阳性胶质瘤干细胞MGMT、NF-κB表达情况。结果与结论:经Western blot检测,胶质瘤干细胞U251G、SKMG-4G中MGMT蛋白呈阳性表达。采用预用干扰素α/β,然后添加替莫唑胺方式,提高了替莫唑胺的化疗敏感性,杀伤效应显著提高。使用干扰素α/β之后,SKMG-4G和U251G MGMT、NF-κB m RNA和蛋白表达均显著降低,联合替莫唑胺之后下降情况更明显。结果表明对于MGMT阳性胶质瘤干细胞,干扰素α/β可以显著增强替莫唑胺的杀伤效应,逆转替莫唑胺的耐药性。BACKGROUND： It is easy to appear the phenomenon of drug resistance in the treatment of patients with glioma, and O（6）-methylguanine-DNA methyltransferase（MGMT） is a kind of multidrug resistance gene expressed in glioma. OBJECTIVE： To investigate the role of interferon to increase the sensibilization of MGMT positive glioma stem cells to temozolomide in vitro. METHODS： Glioma cell lines, U251 and SKMG-4, were induced by suspended cloning ball formation method to harvest MGMT positive glioma stem cells, U251 G and SKMG-4G. Cell counting kit-8 assay was used to detect the killing effect of interferon α/β combined with temozolomide on MGMT positive glioma stem cells. RT-PCR and western blot assay were employed to determine the expression of MGMT and nuclear factor κB in MGMT positive glioma stem cells. RESULTS AND CONCLUSION： Western blot results showed positive expression of MGMT in U251 G and SKMG-4G cells at protein levels. After intervention with interferon α/β, the m RNA expression of MGMT and nuclear factor κB in SKMG-4G and U251 G cells was reduced significantly, and then further decreased after temozolomide treatment. These findings indicate that interferon α/β can remarkably strengthen the killing effect of temozolomide on MGMT positive glioma stem cells.

关 键 词：干细胞 胶质瘤 胶质瘤干细胞 干扰素 替莫唑胺 化疗 耐药性 O6-甲基鸟嘌呤-DNA甲基转移酶 

分 类 号：R394.2[医药卫生—医学遗传学]