CircAKT3对骨肉瘤U2OS细胞增殖和凋亡的影响

Effect of CircAKT3 on the proliferation and apoptosis of osteosarcoma U2OS cells

[目的]通过研究CircAKT3在骨肉瘤U2OS细胞中增殖和凋亡的作用,探讨CircAKT3对骨肉瘤生物学行为的影响。[方法]采用qRT-PCR检测骨肉瘤U2OS细胞和成骨细胞h FOB1.19细胞中CircAKT3的表达水平。后续实验分为两组,实验组U2OS细胞转染CircAKT3的SiRNA,对照组U2OS细胞转染SiRNA对照序列。采用qRTPCR检测,CircAKT3表达CCK-8检测细胞增殖,流式细胞仪检测细胞的凋亡率。[结果]qRT-PCR结果显示,U2OS细胞系中CircAKT3的表达水平明显高于h FOB1.19细胞系(P<0.05)。转染24 h后,实验组中CircAKT3的表达水平较对照组明显降低(P<0.05)。CCK-8实验结果显示,实验组细胞在24、48 h的OD值和对照组无明显差异,而在72、96 h的OD值明显低于对照组(P<0.05)。流式细胞仪检测结果显示,细胞的凋亡率实验组为(11.57±2.82)%,对照组为(2.21±1.17)%,差异有统计学意义(P<0.05)。[结论]骨肉瘤细胞的CircAKT3高表达可能与其肿瘤生物学行为有关。转染CircAKT3的SiRNA序列可以沉默CircAKT3的表达,抑制骨肉瘤U2OS细胞的增殖,并促进骨肉瘤细胞U2OS的凋亡。

[Objective]To explore the role of CircAKT3 on the biological behaviors of osteosarcoma by assay of CircAKT3 expression and the proliferation and apoptosis of osteosarcoma U2 OS cells.[Methods]qRT-PCR assay was conducted to detect the expression of CircAKT3 in osteosarcoma U2 OS cells and osteoblast hFOB1.19 cells.Subsequently,the U2 OS cells were divided into two groups,the cells in the trial group were transfected with SiRNA for CircAKT3,while those in the control group transfected with the SiRNA of control sequence.The expression of CircAKT3 was detected by qRT-PCR,cell proliferation was assessed by CCK-8 assay,and cell apoptosis was measured by flow cytometry.[Results]The expression of CircAKT3 in the U2 OS cells was significantly higher than that in the h FOB1.19 cells(P<0.05).However,the expression of CircAKT3 in the trial group was significantly lower than that in the control group 24 hours after SiRNA transfection.Regarding to cell proliferation,the OD values of CCK-8 assay in the trial group at 72 h and 96 h were significantly lower than those in the control group(P<0.05),although there were no statistically significances at 24 h and 48 h between the two groups(P>0.05).As results of flow cytometry,the apoptosis rate of cells was of(11.57±2.82)%in the trial group,whereas(2.21±1.17)%in the control group,which was of statistical significance(P<0.05).[Conclusion]High expression of CircAKT3 in the U2 OS cells might be related to biological behaver of osteosarcoma.Transfection of SiRNA sequence of CircAKT3 does silence the expression of CircAKT3 in U2 OS cells,inhibit cell proliferation and enhance cell apoptosis.