微小RNA-154-5p通过靶向作用于E2F5对骨肉瘤细胞生物活性的影响及其机制  

Effects and mechanism of microRNA-154-5p targeting E2F5 on bioactivity of osteosarcoma cells

作　　者：李彦华 田青 韩奇财[1] LI Yanhua;TIAN Qing;HAN Qicai(Department of Orthopaedics,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院骨科,河南郑州450052

出　　处：《肿瘤基础与临床》2024年第2期131-134,共4页journal of basic and clinical oncology

摘　　要：目的探讨微小RNA-154-5p(miR-154-5p)靶向E2F5对骨肉瘤细胞增殖和凋亡的影响及其分子机制。方法MiR-154-5p转染骨肉瘤细胞MG63后,CCK-8法检测细胞增殖变化情况,Transwell细胞迁移实验评估细胞迁移及侵袭能力的改变,流式细胞仪检测细胞凋亡的水平,Western blot检测凋亡相关标志性分子B细胞淋巴瘤2蛋白关联X蛋白(Bax)、Cleaved caspase 3、B细胞淋巴瘤2蛋白(Bcl-2)表达的变化,生物信息学技术及双荧光素酶报告基因分析miR-154-5p的潜在靶基因。结果CCK-8检测结果显示,空白组、阴性对照组、miR-154-5p模拟组干预72 h后吸光度值分别为1.179±0.026、1.161±0.024、0.502±0.015,后者明显降低(F=2533.000,P<0.001)。Transwell细胞迁移实验结果显示,空白组、阴性对照组、miR-154-5p模拟组干预72 h后MG63细胞迁移数量分别为79.80±8.53、76.20±6.38、49.60±3.05,后者明显减少(F=33.260,P<0.001)。流式细胞仪检测结果显示,空白组、阴性对照组、miR-154-5p模拟组干预72 h后MG63细胞凋亡率分别为(4.56±1.04)%、(4.25±1.02)%、(18.12±1.90)%,后者明显上升(F=99.400,P<0.001)。Western blot进一步检测发现,空白组、阴性对照组、miR-154-5p模拟组干预72 h后Bax、Cleaved caspase 3、Bcl-2表达明显增加(Bax:0.124±0.010、0.128±0.013、0.364±0.018;Cleaved caspase 3:0.099±0.020、0.112±0.033、0.513±0.024;Bcl-2:0.672±0.015、0.675±0.017、0.245±0.028;F=188.200,P<0.001;F=159.200,P<0.001;F=281.600,P<0.001)。生物信息学和双荧光素酶报告基因分析结果显示,E2F5为miR-154-5p潜在的靶基因。结论MiR-154-5p可以通过抑制E2F5促进骨肉瘤细胞凋亡以及抑制细胞增殖。Objective To investigate the effect of microRNA-154-5p(miR-154-5p)targeting E2F5 on the proliferation and apoptosis of osteosarcoma cells and its molecular mechanism.Methods After miR-154-5p transfection of osteosarcoma MG63 cells,the changes of cell proliferation were detected by CCK-8,the changes of cell migration and invasion ability were evaluated by transwell cell migration assay,the level of apoptosis was detected by flow cytometry.Western blot was used to analyze the changes in the expression of B-cell lymphoma protein 2-associated X protein(Bax),Cleaved caspase-3 and B-cell lymphoma protein 2(Bcl-2)markers.Bioinformatics and dual luciferase reporter genes were used to analyze the potential target genes of miR-154-5p.Results The CCK-8 assay results showed that the absorbance values of the blank group,the negative control group and the miR-154-5p simulated group after 72 hours of intervention were 1.179±0.026,1.161±0.024 and 0.502±0.015,respectively,the difference was statistically significant(F=2533.000,P<0.001).The Transwell cell migration experiment results showed that the number of migrating MG63 cells in the blank group,the negative control group and the miR-154-5p simulated group after 72 hours of intervention were 79.80±8.53,76.20±6.38 and 49.60±3.05,respectively,the difference was statistically significant(F=33.260,P<0.001).The flow cytometry detection results showed that the apoptosis rates of MG63 cells in the blank group,the negative control group and the miR-154-5p simulated group after 72 hours of intervention were(4.56±1.04)%,(4.25±1.02)%and(18.12±1.90)%,respectively,the difference was statistically significant(F=99.400,P<0.001).Western blot further detected that the expression of Bax,Cleaved caspase 3,and Bcl-2 in the blank group,the negative control group and the miR-154-5p simulated group after 72 hours of intervention was significantly increased(Bax:0.124±0.010,0.128±0.013,0.364±0.018;Cleaved caspase 3:0.099±0.020,0.112±0.033,0.513±0.024;Bcl-2:0.672±0.015,0.675±0.017,0.245

关 键 词：骨肉瘤 微小RNA 凋亡 增殖 侵袭 

分 类 号：R738.1[医药卫生—肿瘤]