六氧甲基鸟嘌呤DNA甲基转移酶基因导入增强外周血单个核细胞抗药性

目的研究六氧甲基鸟嘌呤DNA甲基转移酶（MGMT）基因导人后人外周血单个核细胞（PBMC）对烷化剂抗药性的改变。方法利用脂质体法将含人MGMT基因的真核表达载体导人体外培养的外周血单个核细胞，实时PCR及Western blot检测目的基因mRNA及蛋白水平的表达。四氮唑蓝法（MTT）检测细胞对硝卡芥抗药性的改变。结果转染目的基因组在mRNA及蛋白水平与转空载体及对照相比均为高表达。MTT法结果显示转目的基因组对硝卡芥的IC50是转空载体组及对照组的两倍。结论MGMT基因导人能够增强外周血单个核细胞对烷化剂的抗性从而起到保护作用。

Drug-resistance increased by transferring O6-methylguanine-DNA-methyltransferase into peripheral blood mononuclear cells

Purpose To study the change of alkylating agent-resistance in PBMC after MGMT introduction. Methods Mammalian expression vectors containing human MGMT gene were tranferred into PBMC in vitro via liposome. Realtime PCR and Western blot were used to detect the expression of MGMT at mRNA and protein levels. The change of agent-resistance to nitrocaphane was evaluated through MTT. Results Gene-transferred group had distinct higher expression of MGMT at mRNA and protein levels compared with empty vector-transferred group and control group. MTT showed that the IC50 of gene-transferred group was two times that of the other two groups. Conclusions MGMT gene transfection can improve the alkylating agent-resistance in PBMC and therefore perform protective effect.