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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。
Objective To investigate the therapeutic effect of proton sponge QD-siRNA targeting Notch1 protein overexpressed in acute T lymphoblastic leukemia cell line. Methods Acute lymphoblastic leukemia cell line Notch1 was overexpressed in this study. Proton sponge-encapsulated quantum dots (QD) -siRNA complexes targeting Notch1 gene were synthesized and transfected into leukemia cells. The results were compared with that of the traditional transfection reagent Lipofectamine 2000 , Compared with TransIT and JetPEI, the transfection efficiency was detected by Western blotting and RT-PCR, and the cytotoxicity was detected by CCK8 assay. Results Compared with other transfection reagents, the proton sponge-encapsulated QD-siRNA obtained a 4 to 10-fold increase in gene silencing effect and significantly reduced mRNA expression (P <0.01), and its cytotoxicity decreased to other transfection 47.6% ~ 66.7% reagent, the number of viable cells increased 1.5 ~ 2.1 times. Conclusions Proton sponge QD-siRNA can be used in Notch1 acute lymphoblastic leukemia cell lines. Compared with traditional transfection reagent, proton sponge QD-siRNA can enhance gene silencing effect and reduce cytotoxicity.