摘要
目的研究质子海绵量子点-siRNA靶向急性T淋巴细胞白血病细胞系过度表达的Notch1蛋白的治疗效果。方法以Notch1过度表达的急性淋巴细胞白血病细胞系为研究背景,合成靶向Notch1基因的质子海绵包封的量子点(quantum dots,QD)-siRNA复合物转染白血病细胞,与传统转染试剂Lipofectamine2000、TransIT和JetPEI相比较,用免疫印迹方法、RT-PCR方法检测转染效率,用CCK8分析法检测细胞毒作用。结果与其它转染试剂比较,质子海绵包封的QD-siRNA在基因沉默效应上取得了4～10倍的提高,同时mRNA的表达明显减少(P<0.01),其细胞毒性下降到其它转染试剂的47.6%～66.7%,存活细胞数量上升了1.5～2.1倍。结论质子海绵QD-siRNA应用到Notch1急性T淋巴细胞白血病细胞系中,与传统转染试剂比较,可提高基因沉默效应并降低细胞毒作用。
We aimed to research the therapeutic effect of short-interfering RNA （siRNA） of proton-spongecoated quantum dots （QD） to Notchl protein over expression in T cell acute lymphocyte leukemia （T-ALL） cell line. Firstly, proton-sponge-coated QD-siRNA targeting Notchl protein was constructed and transfected into T- ALL cell line, while transfection reagent such as Lipofectamine 2000, TransiT and JetPEI were set as controls. Immunoblotting and RT-PCR indicated that the gene silencing efficiency of proton-sponge-coated QD-siRNA increased by 4-10 folds, and proton-sponge-coated QD-siRNA also decreased the content of Notchl mRNA significantly （P 〈 0.01）, as compared with controls. CCK8 assay showed that proton-sponge-coated QD-siRNA decreased the cellular toxicity to 47.6%-66.7% compared with other reagent but increased the survival rate of T-ALL cell line by 1.5-2.1 folds, compared with other transfection reagent. Above data represents improvement in gene silencing efficiency and reduction in cellular toxicity when proton-sponge-coated QD-siRNA is applied to therapy of T-ALL.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2013年第6期474-479,共6页
Immunological Journal
基金
国家自然科学基金青年基金项目(81100350)
吉林大学基本科研业务费(450060445226)
第49批中国博士后科学基金面上资助一等资助金(20110490155)
吉林大学白求恩B计划资助基金(450060481921)
