lncRNA AC131056.3通过调控miR-21-5p对骨肉瘤细胞增殖和侵袭的影响  

Effects of lncRNA AC131056.3 on proliferation and invasion of osteosarcoma cells by regulating miR-21-5p

作　　者：牟健雄 丁崇军 唐荣慧 叶勇衡 谢琳[2] 李衡 MU Jianxiong;DING Chongjun;TANG Ronghui;YE Yongheng;XIE Lin;LI Heng(Department of Orthopedics,Xichang People’s Hospital,Xichang 615000,China;Department of Orthopedics,First Affiliated Hospital of Kunming Medical University)

机构地区：[1]四川省西昌市人民医院骨科,西昌615000 [2]昆明医科大学第一附属医院骨科

出　　处：《山西医科大学学报》2022年第8期969-974,共6页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81760520)。

摘　　要：目的探讨长链非编码RNA(long non-coding RNA,lncRNA)AC131056.3对骨肉瘤细胞增殖和侵袭的作用及可能机制。方法采用GEPIA数据库分析骨肉瘤患者总生存期和AC131056.3表达的相关性。采用qRT-PCR检测骨肉瘤细胞(Saos2、U-2OS、MG-63、143B、HOS)及正常成骨细胞hFOB1.19中AC131056.3的表达。将HOS细胞分为对照组和AC131056.3组,分别转染pcDNA质粒和pcDNA-AC131056.3质粒。CCK-8法检测HOS细胞增殖活力,Transwell实验检测HOS细胞侵袭活力,qRT-PCR检测两组HOS细胞中miR-21-5p表达,Western blot检测两组HOS细胞中PI3K/Akt/mTOR信号通路蛋白p-Akt、p-mTOR、p-PI3K、GSK-3、PDK-1表达。生物信息学和双荧光素酶报告基因法验证AC131056.3与miR-21-5p的互补关系。结果与AC131056.3低表达的骨肉瘤患者相比,AC131056.3高表达的患者总生存期较长(P<0.01)。与hFOB1.19细胞比较,骨肉瘤细胞Saos2、U-2OS、MG-63、143B、HOS中AC131056.3表达降低(P<0.05),HOS细胞中AC131056.3的表达最低(P<0.01)。与对照组比较,AC131056.3组HOS细胞增殖活力显著降低(P<0.05),细胞侵袭活力显著降低(P<0.01)。AC131056.3可靶向结合miR-21-5p(P<0.01)。与对照组比较,AC131056.3组HOS细胞中miR-21-5p表达降低(P<0.01),PI3K/Akt/mTOR信号通路蛋白p-Akt、p-mTOR、p-PI3K、GSK-3、PDK-1表达降低。结论AC131056.3表达与骨肉瘤患者总生存期正相关,AC131056.3可通过靶向抑制miR-21-5p的表达降低HOS细胞的增殖及侵袭活力。Objective To investigate the effect of long non-coding RNA(lncRNA)AC131056.3 on the proliferation and invasion of osteosarcoma cells and its possible mechanism.Methods The GEPIA database was used to analyze the correlation between OS and AC131056.3 expression in patients with osteosarcoma.The expression of AC131056.3 was detected in osteosarcoma cells Saos2,U-2OS,MG-63,143B,HOS and normal osteoblast hFOB1.19 by qRT-PCR.HOS cells were divided into control group and AC131056.3 group,and were transfected with pcDNA plasmid and pcDNA-AC131056.3 plasmid respectively.The proliferation activity of HOS cells was detected by CCK-8 assay,and the invasion activity of HOS cells was detected by Transwell assay.The expression of miR-21-5p in HOS cells was detected in the two groups by qRT-PCR.Western blot was used to detect the expression of PI3K/Akt/mTOR signaling pathway proteins p-Akt,p-mTOR,p-PI3K,GSK-3 and PDK-1 in HOS cells in two groups.The complementary relationship between AC131056.3 and miR-21-5p was verified by bioinformatics technology and dual luciferase reporter gene method.Results Compared with osteosarcoma patients with low expression of AC131056.3,the overall survival was longer in patients with high expression of AC131056.3(P<0.01).Compared with hFOB1.19 cells,the expression of AC131056.3 in osteosarcoma cells Saos2,U-2OS,MG-63,143B and HOS was decreased(P<0.05),and the expression of AC131056.3 in HOS cells was the lowest(P<0.01).Compared with control group,the proliferation activity of HOS cells in AC131056.3 group was significantly decreased(P<0.05),and the cell invasion activity was significantly decreased(P<0.01).AC131056.3 could target and bind to miR-21-5p(P<0.01).Compared with control group,the expression of miR-21-5p in HOS cells in AC131056.3 group was decreased(P<0.01),and the expression of PI3K/Akt/mTOR signaling pathway proteins p-Akt,p-mTOR,p-PI3K,GSK-3 and PDK-1 was decreased.Conclusion The expression of AC131056.3 was positively correlated with the overall survival of patients with osteosarcoma

关 键 词：骨肉瘤 长链非编码RNA miR-21-5p 细胞增殖 细胞侵袭 

分 类 号：R738.1[医药卫生—肿瘤]