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目的:研究了丁胱亚磺酰亚胺(BSO)增强D-氨基酸氧化酶(DAAO)/D-丙氨酸(D-Ala)自杀基因系统在白血病基因治疗中的作用。方法:应用逆转录病毒转染技术获得稳定表达DAAO的高致瘤性K562e单克隆细胞K_(DIGC),用PCR、原位杂交对DAAO基因修饰的K_(DfGC)进行鉴定。应用MTT法检测D-Ala对BSO预处理过的K_(DfGC)细胞的杀伤作用。结果:PCR和原位杂交分析证明DAAO基因已整合至细胞基因组中,并在mRNA水平上表达。D-Ala作用24h,K_(DfGC)细胞的半数抑制浓度(C_(50))为10.21mmol/L,K_(DfGC)+BSO的IC_(50)为7.92mmol/L,两者的95％可信区间不重叠。结论:BSO可增强DAAO/D-Ala系统杀伤K652e细胞作用。
OBJECTIVE: To investigate the role of the BSA-enhanced DAAO / D-Ala suicide gene system in the gene therapy of leukemia. Methods: The KAO (DIGC), a monoclonal antibody against K562e cell line stably expressing DAAO, was obtained by retroviral transfection. The DAAO gene modified K_ (DfGC) was identified by PCR and in situ hybridization. The killing effect of D-Ala on BSF pretreated K_ (DfGC) cells was detected by MTT assay. Results: PCR and in situ hybridization analysis showed that the DAAO gene has been integrated into the cell genome and expressed at the mRNA level. The half inhibitory concentration (C_ (50)) of K_ (DfGC) cells was 10.21 mmol / L and the IC_ (50) of K_ (DfGC) + BSO was 7.92 mmol / L after D-Ala treatment for 24 h. The confidence intervals do not overlap. Conclusion: BSO can enhance the killing effect of DAAO / D-Ala on K652e cells.