环状RNA circAKT3对骨肉瘤细胞生物学行为的影响

Effect of circular RNA circAKT3 on the biological behaviors of osteosarcoma cells

目的通过研究环状RNA circAKT3对骨肉瘤细胞生物学行为的影响,来明确circAKT3在骨肉瘤进展中的作用。方法将骨肉瘤细胞分为实验组和对照组,实验组转染circAKT3的siRNA序列,对照组转染siRNA NC(negative control)序列。采用qRT-PCR验证转染效果,后续采用CCK-8实验检测两组细胞的增殖能力,采用流式细胞仪检测两组细胞的凋亡率,采用Transwell侵袭和迁移实验分别检测两组细胞的侵袭和迁移能力。采用qRT-PCR检测患者骨肉瘤及瘤旁组织中circAKT3的表达水平,然后分析骨肉瘤组织中circAKT3的表达水平与骨肉瘤患者临床相关病理参数的相关性。结果qRT-PCR检测结果显示,实验组SOSP-9607细胞中circAKT3的表达水平较对照组明显降低(P<0.001)。CCK-8实验和流式细胞仪凋亡检测结果显示,实验组SOSP-9607细胞的增殖能力较对照组明显降低(P<0.05),而凋亡率则明显升高(P<0.001)。Transwell结果显示,实验组SOSP-9607和U2OS细胞的侵袭和迁移能力较对照组明显降低(P<0.01)。骨肉瘤组织内circAKT3的表达水平较瘤旁组织明显升高(P<0.01)。骨肉瘤组织内circAKT3的表达水平与患者的临床分期和转移呈正相关(P<0.05)。结论circAKT3可以增强骨肉瘤细胞的恶性生物学行为,促进骨肉瘤的进展。

Objective To explore the effect of circular RNA circAKT3 on the biological behaviors of osteosarcoma cells,and identify the role of circAKT3 in the progression of osteosarcoma.Methods The osteosarcoma cells were divided into experimental group(transfected with siRNA sequence against circAKT3)and control group(transfected with negative control sequence of siRNA).Subsequent transfection efficiency was confirmed by qRT-PCR.Then,CCK-8 assay,flow cytometry analysis and Transwell assay were adopted to detect the proliferation,the apoptosis rate,the invasion and migration abilities in the two groups,respectively.The expression of circAKT3 was detected in osteosarcoma and adjacent normal tissues from patients by qRT-PCR.The correlation between circAKT3 expression and clinically-related pathological parameters of patients with osteosarcoma was analyzed.Results The qRT-PCR result showed the expression of circAKT3 in SOSP-9607 cells was significantly decreased in experimental group compared to control group(P<0.001).Compared with control group,the proliferation ability of SOSP-9607 cells in experimental group was significantly reduced by CCK-8 assay(P<0.05),while the apoptosis rate of SOSP-9607 cells in experimental group was significantly increased by flow cytometry analysis(P<0.001).The invasion and migration abilities of SOSP-9607 and U2OS cells in experimental group were significantly lower than those in control group by Transwell assay(P<0.01).The expression of circAKT3 in osteosarcoma tissues was higher than that in adjacent normal tissues(P<0.01).Moreover,the expression of circAKT3 was positively correlated with clinical stage and metastasis of patients with osteosarcoma(P<0.05).Conclusion The circAKT3 enhances the malignant biological behaviors of osteosarcoma cells and thus promotes the progression of osteosarcoma.