中国当代儿科杂志2005，Vol.7Issue(3)：202-206,5.
RNAi阻断NPM-ALK基因表达及对大细胞间变性淋巴瘤细胞的影响
Effects of RNA interference on NPM-ALK fusion gene expression in anaplastic large-cell lymphoma cells
摘要
Abstract
Objective To evaluate two small interfering RNAs (siRNAs) on the NPM-ALK fusion gene expression in anaplastic large-cell lymphoma cell line Karpas299, and to study the effect of RNA interference on Karpas299 cells proliferation. Methods Two siRNAs sequences (siRNA- I and siRNA-II) were designed to target the NPM-ALK fusion site in anaplastic large-cell lymphoma cell line Karpas299. An siRNA U6 expression system including U6 RNA-based polymerase III promoter was set up. The two siRNAs designed for down-regulation of the NPM-ALK fusion mRNA were transfected into Karpas299 cells by liposomal transfection reagents. The effect of RNAi on NPM-ALK mRNA expression was detected by real-time RT-PCR and Western blot. The anti-proliferative effects of the siRNA U6 system were assessed using MTT. Apoptosis was observed by fluorescence microscopy. Results The mRNA level of NPM-ALK in Karpas299 cells transfected with siRNA-I and siRNA-II decreased by approximately 75% and 35% respectively. The NPM-ALK protein expression was inhibited in Karpas299 cells at 72 hrs of siRNA-I transfection. The siRNA-II treatment had no effect on NPM-ALK protein expression. siRNA-I had inhibitory effects on Karpas299 cells proliferation and induced the cells apoptosis, while siRNA-II did not. Conclusions Sequence specific siRNAs targeting NPM-ALK was capable of suppressing NPM-ALK expression and inhibiting cellular proliferation. RNA interference may be a suitable technique for studying the function of NPM-ALK gene and may be used to develop siRNA-based targeted gene therapeutic approaches against NPM-ALK-positive lymphomas.关键词
RNA干扰/siRNA/融合基因/NPM-ALK/大细胞间变性淋巴瘤Key words
RNA interference/RNA, small interfering/Gene fusion/NPM-ALK/Anaplastic large-cell lymphoma分类
医药卫生引用本文复制引用
赵艳霞,顾龙君,叶启东,赵金彩..RNAi阻断NPM-ALK基因表达及对大细胞间变性淋巴瘤细胞的影响[J].中国当代儿科杂志,2005,7(3):202-206,5.基金项目
Supported by the Technology Commission of Shanghai Municipal Government of China under Grant No. 02DJ14011. （）
