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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 short hairpin RNA (shRNA) on the proliferation and apoptosis of osteosarcoma cell line SOSP-9607. METHODS: Cy-clin D1 shRNA expression vector was used to stably transfect the osteosarcoma cell line SOSP-9607. The changes of CyclinD1 mRNA and protein were detected by RT-PCR and Western blot respectively. The expressions of osteosarcoma The changes of cell cycle and apoptosis rate of CyclinD1 were observed. The effect of Cyclin D1 on the proliferation of osteosarcoma cells was detected by CCK-8. Results: The expression of Cyclin D1 mRNA and protein in osteosarcoma cells was significantly inhibited after stable transfection of Cyclin D1 shRNA vector. Compared with the control group, proliferation of osteosarcoma cells was significantly inhibited (P <0.05). At the same time, G0 / 1 arrest occurred in the cells and the proportion of G1 / S phase increased (P <0.01). Conclusion: Cyclin D1 shRNA vector can down-regulate the expression of the target gene and effectively inhibit the proliferation of SOSP-9607 osteosarcoma cell line, resulting in G1 / S arrest.