circ_0001461靶向抑制miR-30a-5p对骨肉瘤细胞增殖和凋亡的影响  

The Effect of Circ_0001461 Targeting Inhibition of miR-30a-5p on the Proliferation and Apoptosis of Osteosarcoma Cells

作　　者：范金柱 从飞 张文韬 田小宁 宋涛[1] 郭云山 FAN Jin-zhu;CONG Fei;ZHANG Wen-tao;TIAN Xiao-ning;SONG Tao;GUO Yun-shan(Department of Bone Microsurgery,Honghui Hospital,Xi'an Jiaotong University,Xi'an,Shaanxi,710054,China;Department of Spinal Surgery,Honghui Hospital,Xi'an Jiaotong University,Xi'an,Shaanxi,710054,China)

机构地区：[1]西安交通大学附属红会医院骨显微修复外科,陕西西安710054 [2]西安交通大学附属红会医院脊柱外科,陕西西安710054

出　　处：《现代生物医学进展》2022年第8期1413-1418,共6页Progress in Modern Biomedicine

基　　金：西安交通大学基本科研业务费(xzy01201923);陕西省2020年自然科学基础研究计划(2020JM-687)。

摘　　要：目的:探讨circ_0001461对骨肉瘤细胞增殖和凋亡的影响及调控机制。方法:采用实时荧光定量聚合酶反应(qRT-PCR)检测检测circ_0001461在骨肉瘤组织和细胞中的表达水平。在U2OS和HOS细胞中转染sh-NC和sh-circ_0001461后,采用CCK8检测细胞增殖情况,流式细胞术检测细胞凋亡情况,qRT-PCR检测增殖相关分子Ki-67 m RNA的表达水平,Western Blot检测凋亡相关分子Cleaved-caspase-3蛋白的表达水平。采用双荧光素酶报告基因检测circ_0001461和mi R-30a-5p的结合情况。结果:circ_0001461在骨肉瘤组织中的表达水平明显高于癌旁正常组织(P<0.05),circ_0001461在骨肉瘤细胞U2OS和HOS中的表达水平均明显高于成骨细胞NHOst(P<0.05)。低表达circ_0001461能够抑制骨肉瘤细胞U2OS和HOS的增殖和增殖相关分子Ki-67的表达(P<0.05);促进骨肉瘤细胞U2OS和HOS的凋亡和凋亡相关分子Cleaved-caspase-3蛋白的表达(P<0.05)。双荧光素酶结果显示circ_0001461能够靶向结合mi R-30a-5p。低表达circ_0001461能够促进mi R-30a-5p的表达(P<0.05),circ_0001461和mi R-30a-5p在骨肉瘤组织中的表达呈负相关(P<0.05)。在U2OS细胞中共转染sh-circ_0001461和mi R-30a-5p mimics后能够进一步加强单独转染sh-circ_0001461对U2OS细胞增殖和凋亡的影响(P<0.05);在HOS细胞中共转染sh-circ_0001461和mi R-30a-5p inhibitors后能够逆转单独转染sh-circ_0001461对U2OS细胞增殖和凋亡的影响(P>0.05)。结论:circ_0001461在骨肉瘤组织和细胞中明显高表达,低表达circ_0001461能够靶向促进mi R-30a-5p的表达进而抑制骨肉瘤细胞增殖和促进细胞凋亡。Objective:To investigate the effect of circ_0001461 on the proliferation and apoptosis of osteosarcoma cells and its regulatory mechanism.Methods:Real-time fluorescent quantitative polymerase reaction(qRT-PCR)was used to detect the expression level of circ_0001461 in osteosarcoma tissues and cells.After transfection of sh-NC and sh-circ_0001461 in U2OS and HOS cells,CCK8was used to detect cell proliferation,flow cytometry was used to detect cell apoptosis,qRT-PCR was used to detect the expression level of proliferation-related molecule Ki-67 m RNA,Western Blot was used to detect the expression level of apoptosis-related molecule Cleaved-caspase-3 protein.The dual luciferase reporter gene was used to detect the binding of circ_0001461 and mi R-30a-5p.Results:The expression level of circ_0001461 in osteosarcoma tissue was significantly higher than that in normal adjacent tissues(P<0.05),and the expression level of circ_0001461 in osteosarcoma cells U2OS and HOS was significantly higher than that in osteoblast NHOst(P<0.05).Low expression of circ_0001461 could inhibit the proliferation of osteosarcoma cells U2OS and HOS and the expression of proliferation-related molecule Ki-67(P<0.05);promote the apoptosis of osteosarcoma cells U2OS and HOS and the apoptosis-related molecule Cleaved-caspase-3 protein Expression(P<0.05).The results of dual luciferase showed that circ_0001461 could target mi R-30a-5p.Low expression of circ_0001461 could promote the expression of mi R-30a-5p(P<0.05),and the expression of circ_0001461 and mi R-30a-5p in osteosarcoma tissues was negatively correlated(P<0.05).Co-transfection of sh-circ_0001461 and mi R-30a-5p mimics in U2OS cells could further enhance the effect of single transfection of sh-circ_0001461 on the proliferation and apoptosis of U2OS cells(P<0.05);co-transfection of sh-circ_0001461 and mi R-30a-5p inhibitors in HOS cells could reverse the effect of single transfection sh-circ_0001461 on the proliferation and apoptosis of U2OS cells(P>0.05).Conclusion:circ_0001461 was highly e

关 键 词：骨肉瘤 circ_0001461 mi R-30a-5p 增殖 凋亡 

分 类 号：R-33[医药卫生] R738.1