摘要
为了探讨以bcl2基因作为靶分子进行白血病多药耐药基因治疗的可行性,构建bcl2基因的小干扰RNA载体并将其转导入HL60/VCR细胞,在G418筛选后应用Westernblot进行鉴定;MTT法检测细胞转染前后生长速度和药物敏感性的变化;Westernblot检测细胞转染前后凋亡相关蛋白Bax和耐药相关蛋白ZNRD1的表达变化。结果表明:成功构建了bcl2的小干扰RNA载体并转染HL60/VCR;经蛋白质水平检测证实成功建立了bcl2低表达的白血病细胞模型;转染后的细胞对长春新碱和阿霉素的敏感性增强,且低表达耐药相关蛋白ZNRD1。结论:bcl2小干扰RNA真核表达载体能在一定程度上逆转白血病细胞的耐药性。
To evaluate the feasibility of gene therapy using bcl-2 as target in multiple drug resistance of leukemia, the small interfering RNA eukaryotic expression vector specific to human bcl-2 gene was constructed by gene recombination, then transfected into HL-60/VCR cells. Stable transfectants were obtained by G418 screening, The growth curve and drug sensitivity were detected by using MTT. The expression of Bax and ZNRD1 was analyzed by Western blot. The results showed that mU6pro-bcl-2 siRNA was successfully constructed and transfected into HL-60/VCR cells. The IC50 of transfected cells to vincristine and adriarnycin was significantly reduced as compared with that of the control. The expression of ZNRD1 in transfected cells was decreased as compared with that of the control, while Bax not. It is concluded that the bcl-2 siRNA restores the sensitivity of HL-60/VCR cells to conventional chemotherapeutic agents to a certain degree.
出处
《中国实验血液学杂志》
CAS
CSCD
2005年第6期1010-1013,共4页
Journal of Experimental Hematology
基金
沈阳军区总医院青年启动基金(No.03YC06)
国家自然科学基金资助项目(No.30370599)
