Lü Jia-Yin, Gao Zhong-Li, Wang Jin-Cheng, Wu Dan-Kai, Zhao Dong-Xu, Zhang Duo-Duo, Zhao Yan-Ying
Department of Orthopedic Surgery, China-Japan Union Hospital of Jilin University, Changchun 130031, China.
Zhonghua Zhong Liu Za Zhi. 2008 Jul;30(7):502-5.
To investigate the inhibitory effect of the small interfering RNA targeting mdm2 gene on the growth of osteosarcoma cells.
PGCsilencerTM-mdm2 siRNA was constructed and transfected into the osteosarcoma cell line U2OS cells. The inhibitory effects on mdm2 were determined by RT-PCR and Western blot analysis. The cell growth activity was determined by MTT assay, and the cell apoptosis was examined by flow cytometry. The therapeutic effects of simdm2 was assessed on the nude mouse model of transplanted tumor.
The simdm2 plasmid was successfully constructed. After simdm2 being transfected into the U2OS cells, the expressions of mdm2 gene and protein were significantly inhibited. The ability of cell growth activity decreased greatly and cell apoptosis occurred apparently. There was no significant difference between the negative control group and non-transfected group. The growth of xenograft tumor in simdm2 transfected nude mice was inhibited and the expressions of mdm2 gene and protein were down-regulated remarkably.
siRNA targeting mdm2 gene inhibits the mdm2 expression in osteosarcoma U2OS cells and the growth of osteosarcoma in nude mice.
探讨靶向mdm2基因的小干扰RNA对骨肉瘤细胞生长的抑制作用。
构建PGCsilencerTM-mdm2 siRNA并转染至骨肉瘤细胞系U2OS细胞。通过RT-PCR和蛋白质印迹分析确定对mdm2的抑制作用。采用MTT法测定细胞生长活性,通过流式细胞术检测细胞凋亡。在移植瘤裸鼠模型上评估simdm2的治疗效果。
成功构建了simdm2质粒。将simdm2转染至U2OS细胞后,mdm2基因和蛋白的表达受到显著抑制。细胞生长活性能力大幅下降,细胞凋亡明显发生。阴性对照组与未转染组之间无显著差异。转染simdm2的裸鼠体内异种移植瘤的生长受到抑制,mdm2基因和蛋白的表达显著下调。
靶向mdm2基因的siRNA抑制骨肉瘤U2OS细胞中mdm2的表达及裸鼠体内骨肉瘤的生长。