ShRNA表达载体干涉Cyclin D1分子抑制骨肉瘤细胞系SOSP-9607的增殖

ShRNA of Cyclin D1 decreased the proliferation of human osteosarcoma cell line SOSP-9607

目的:探讨Cyclin D1 shRNA(short hairpin RNA)对骨肉瘤细胞系SOSP-9607增殖和凋亡的影响。方法:利用Cy-clin D1 shRNA表达载体,稳定转染骨肉瘤细胞系SOSP-9607,分别采用RT-PCR和Western blot分别从mRNA水平和蛋白水平上检测CyclinD1的变化;流式细胞术检测骨肉瘤细胞Cyc-linD1周期和凋亡率的变化;CCK-8检测干涉Cyclin D1对骨肉瘤细胞增殖的影响。结果:稳定转染Cyclin D1 shRNA载体后骨肉瘤细胞的Cyclin D1 mRNA和蛋白的表达均被明显抑制。与对照组相比,骨肉瘤细胞增殖被显著抑制(P<0.05)。同时,细胞发生G0/1期阻滞,G1/S期比例增加(P<0.01)。结论:Cyclin D1 shRNA载体可下调目的基因的表达,有效的抑制骨肉瘤细胞SOSP-9607的增殖,发生G1/S期阻滞。

AIM： To investigate the effect of Cyclin D1 shRNA on the apoptosis and proliferation of human osteosarcoma cell line SOSP-9607. METHODS： Human Cyclin D1 shRNA vector was stably transfected into SOSP-9607 osteosarcoma cells. The mRNA and protein cxpression levels of Cyclin DI were detected by semiquantitative RT-PCR and Western blot respectively. The cell cycle and pretiferation of osteosarcoma cells were examined by FCM analysis and CCK-8 method, respectively. RESULTS： After stable transfection of Cyclin D1 shRNA, the expression of Cyclin DI were inhibited at mRNA and protein levels. The proliferation of SOSP-9607 osteosarcoma cells was inhibited. The difference was significant compared with control groups （P 〈 0.05）. At the same time, Cyclin D1 shRNA transfection increased G0/1 phage content and decreased S phage content. CONCLUSION： Cyclin D1 shRNA could down-regulate the expression of Cyclin D1, effectively inhibit the proliferation of osteosarcoma cells, and have significant effect on the cell cycle.