shRNA干扰COPS3基因对人成骨肉瘤细胞的影响

Effect of shRNA interfering COPS3 gene on human osteosarcoma cell line

目的:探究shRNA干扰COPS3基因对人成骨肉瘤细胞基因水平、蛋白水平、增殖能力等方面的影响。方法:选择人成骨肉瘤Saos-2细胞,感染pLKO.1-COPS3shRNA病毒质粒作为Saos-2siCOPS3细胞(实验组),感染pLKO.1-TRC control病毒质粒作为Saos-2CON细胞(对照组),感染成功后进行细胞培养,收集细胞用RT-PCR测定COPS3基因mRNA表达量,Western blot检测蛋白表达水平,MMT测定细胞增殖能力,对裸鼠接种细胞悬液进行裸鼠成瘤试验。结果:Saos-2siCOPS3细胞中,COPS3基因mRNA表达水平与对照组相比下降(P<0.05),蛋白表达水平、细胞增殖能力与对照组相比下降(P<0.05),干扰COPS3基因与对照组相比抑制肿瘤体积、重量增加(P<0.05)。结论:COPS3调控骨肉瘤细胞的增殖与骨肉瘤的发生、发展密切相关。

Objective： This paper evaluated the effect of shRNA interference COPS3 gene on the gene, protein level and proliferation ability of human osteosarcoma cells. Methods： Human osteosarcoma Saos-2 cells were select-ed in this study, the cells which infected shRNA pLKO. 1-COPS3 virus plasmid were siCOPS3 Saos-2（treatment）, meanwhile, which infected pLKO. 1-TRC control virus plasmid were CON Saos-2（control）. After infection comple-ted ,cells was cultured then collected for further analyzing. RT-PCR was used to determine mRNA expression of COPS3 gene, western blot was used to determine the level of protein expression, cell proliferation ability was meas-ured by MMT method, nude mice were inoculated with cell suspension to show tumor formation. Results： In si- COPS3 Saos-2 cells, the mRNA expression level of COPS3 was decreased （P〈0. 05） , The level of protein expres-sion, cell proliferation ability decreased, the interference of COPS3 gene inhibited tumor volume and weight increase （P〈0 . 05） compared with the control group. Conclusion： The results of this study suggest COPS3 regulates the pro-liferation of osteosarcoma cells, and is closely related to the occurrence and development of osteosarcoma.