lncRNA SNHG3靶向调控miR-514a-5p对骨肉瘤细胞增殖、凋亡、迁移和侵袭的影响  

Effect of lncRNA SNHG3 on the proliferation, apoptosis, migration and invasion of osteosarcoma cells by targeting miR-514a-5p

作　　者：谭晓谦[1] 梅海波[1] 伍江雁[1] 严安[1] TAN Xiaoqian;MEI Haibo;WU Jiangyan;YAN An(Department of Orthopaedics,Hu’nan Children’s Hospital,Hu’nan Province,Changsha410007,China)

机构地区：[1]湖南省儿童医院骨科,湖南长沙410007

出　　处：《中国医药导报》2021年第22期22-27,共6页China Medical Herald

基　　金：湖南创新型省份建设专项项目(2019SK4006);湖南省儿童医院院级科研课题(20203716B)。

摘　　要：目的探讨长链非编码RNA核仁小分子RNA宿主基因3(SNHG3)对骨肉瘤细胞增殖、凋亡、迁移和侵袭的影响及其作用机制。方法qRT-PCR检测成骨细胞hFOB1.19和骨肉瘤细胞SW-1353、HOS、U-2OS、Saos-2中SNHG3和miR-514a-5p表达。取对数生长期的U-2OS细胞进行转染,分为si-NC组(转染si-NC)、si-SNHG3组(转染si-SNHG3)、miR-NC组(转染miR-NC)、miR-514a-5p组(转染miR-514a-5p)、si-SNHG3+anti-miR-NC组(共转染si-SNHG3和anti-miR-NC)和si-SNHG3+anti-miR-514a-5p组(共转染si-SNHG3和anti-miR-514a-5p)。CCK-8法和流式细胞术检测细胞的活性与凋亡情况;Western blot法检测c-Myc、Bax、MMP-2蛋白表达情况;Transwell小室实验检测细胞迁移和侵袭情况;双荧光素酶实验分析SNHG3与miR-514a-5p的靶向关系。结果骨肉瘤细胞SW-1353、HOS、U-2OS、Saos-2中SNHG3表达高于成骨细胞hFOB1.19,miR-514a-5p表达低于成骨细胞hFOB1.19,差异有统计学意义(P<0.05)。si-SNHG3组的SNHG3表达、细胞活力、迁移细胞、侵袭细胞及c-Myc、MMP-2蛋白表达低于si-NC组;细胞凋亡率、Bax蛋白表达高于si-NC组,差异有统计学意义(P<0.05)。miR-514a-5p组的miR-514a-5p表达、细胞凋亡率、Bax蛋白表达高于miR-NC组;细胞活力、迁移细胞、侵袭细胞及c-Myc、MMP-2蛋白表达低于miR-NC组,差异有统计学意义(P<0.05)。miR-514a-5p组SNHG3野生型荧光素酶活性低于miR-NC组,差异有统计学意义(P<0.05);miR-514a-5p组与miR-NC组SNHG3突变型荧光素酶比较,差异无统计学意义(P>0.05)。si-SNHG3+anti-miR-514a-5p组的miR-514a-5p表达、细胞凋亡率、Bax蛋白表达低于si-SNHG3+anti-miR-NC组,而细胞活力、迁移细胞、侵袭细胞及c-Myc、MMP-2蛋白表达高于si-SNHG3+anti-miR-NC组,差异有统计学意义(P<0.05)。结论敲低SNHG3对骨肉瘤细胞的增殖、侵袭、迁移均有较为明显的抑制作用,同时也可促进骨肉瘤细胞凋亡,其作用机制可能与miR-514a-5p有关。Objective To explore the effect of long non-coding RNA small nucleolar RNA host gene 3(SNHG3)on the proliferation,apoptosis,migration and invasion of osteosarcoma cells and its mechanism.Methods The expression of SNHG3 and miR-514a-5p in osteoblasts hFOB1.19 and osteosarcoma cells SW-1353,HOS,U-2OS,and Saos-2 were detected by qRT-PCR.U-2OS cells in logarithmic growth phase were transfected.They were divided into si-NC group(transfected si-NC),si-SNHG3 group(transfected si-SNHG3),miR-NC group(transfected miR-NC),miR-514a-5p group(transfected miR-514a-5p),si-SNHG3+anti-miR-NC group(co-transfected si-SNHG3 and anti-miR-NC)and si-SNHG3+anti-miR-514a-5p group(co-transfected with si-SNHG3 and anti-miR-514a-5p).CCK-8 assay and flow cytometry were used to check cell viability and apoptosis.The expression of c-Myc,Bax,and MMP-2 protein were detected by Western blot.Cell migration and invasion were tested by Transwell chamber experiment.The target relationship between SNHG3 and miR-514a-5p was analyzed by dual luciferase experiment.Results The expression of SNHG3 in osteosarcoma cells SW-1353,HOS,U-2OS,and Saos-2 were higher than those of osteoblasts hFOB1.19,and the expression of miR-514a-5p were lower than that of osteoblasts hFOB1.19,and the differences were statistically significant(P<0.05).The SNHG3,cell viability,migrating cells,invasive cells,and the expression of c-Myc,MMP-2 protein in si-SNHG3 group were lower than those in si-NC group;while the apoptosis rate and the expression of Bax protein were higher than those in si-NC group,and the differences were statistically significant(P<0.05).The miR-514a-5p expression,apoptosis rate,and the expression of Bax protein in miR-514a-5p group were higher than those in miR-NC group;while cell viability,migrating cells,invasive cells,the expression of c-Myc,MMP-2 protein were lower than those of miR-NC group,and the differences were statistically significant(P<0.05).SNHG3 wild type luciferase activity of miR-514a-5p group was lower than that of miR-NC group,and the differenc

关 键 词：骨肉瘤细胞 增殖 凋亡 迁移 侵袭 

分 类 号：R738.1[医药卫生—肿瘤]