LncRNA XIST通过miR-193a-3p/RSF1轴对骨肉瘤细胞增殖和迁移的作用研究  

Effect of lncRNA XIST on proliferation and migration of osteosarcoma cells through miR-193a-3p/RSF1 axis

作　　者：孔德海[1] 冯德香[2] 刘峻滔 邓明明 付炳金[1] 尹刚[1] 朱晓东[1] KONG Dehai;FENG Dexiang;LIU Juntao;DENG Mingming;FU Bingjin;YIN Gang;ZHU Xiaodong(Department of Foot and Ankle Surgery;Department of Oncology,Affiliated Hospital of Binzhou Medical College,Binzhou,Shandong 256600,China;Department of Traumatology and Orthopedics,Yantai Affiliated Hospital of Binzhou Medical College,Binzhou,Shandong 256600,China)

机构地区：[1]滨州医学院附属医院足踝外科,山东滨州256600 [2]滨州医学院附属医院肿瘤科,山东滨州256600 [3]滨州医学院烟台附属医院创伤骨科,山东滨州256600

出　　处：《重庆医学》2022年第2期198-203,共6页Chongqing medicine

基　　金：山东省医药卫生科技发展计划项目(2017WS753)。

摘　　要：目的研究长链非编码RNA X染色体失活特异转录本(lncRNA XIST)通过miR-193a-3p/重构剪切因子1(RSF1)轴对骨肉瘤细胞增殖、迁移的作用。方法收集2017年9月至2018年12月滨州医学院烟台附属医院30例行骨肉瘤切除术获得的癌组织和癌旁组织,RT-qPCR检测癌组织和癌旁组织中lncRNA XIST mRNA表达。采用脂质体转染法将sh-lncRNA XIST重组质粒、空载质粒分别转至对数期生长的骨肉瘤HOS细胞,另取未做任何处理的HOS细胞为空白组。稳定转染48 h后,CCK-8法检测各组细胞培养24、48、72 h时的吸光度(A)值,划痕实验检测培养24 h时细胞的融合距离,荧光素酶报告基因实验检测LncRNA XIST、miR-193a-3p与RSF1之间的作用关系,RT-qPCR检测lncRNA XIST、miR-193a-3p、RSF1 mRNA表达,Western blot检测RSF1蛋白表达。结果骨肉瘤组织中lncRNA XIST mRNA表达明显高于癌旁组织,差异有统计学意义(P<0.05)。空白组、空载组及实验组A值随转染时间的延长而升高,差异有统计学意义(P<0.05)。与空白组和空载组比较,实验组培养24、48、72 h后A值降低,融合距离缩短,lncRNA XIST、RSF1 mRNA表达降低,miR-193a-3p mRNA表达升高,RSF1蛋白表达降低,差异有统计学意义(P<0.05)。结论敲降lncRNA XIST可抑制骨肉瘤细胞增殖和迁移能力,可能通过miR-193a-3p/RSF1轴发挥作用。Objective To study the effect of long-chain non-coding RNA X chromosome inactivation specific transcripts(lncRNA XIST)on the proliferation and migration of osteosarcoma cells(HOS)through micro RNA-193a-3p(miR-193a-3p)/remodeling and spacing factor1(RSF1)axis.Methods The cancer tissues and paracancerous tissues were collected from 30 cases of osteosarcoma resection in the Yantai Affiliated Hospital of Binzhou Medical College from September 2017 to December 2018.The real-time quantitative PCR(RT-qPCR)was used to detect the expression of lncRNA XIST in cancer tissues and paracancerous tissues.The liposome transfection method was used to transfer the sh-lncRNA XIST recombinant plasmid and unloaded plasmid to logarithmic growth osteosarcoma HOS cells,other HOS cells without any treatment were taken as the blank group.After 48 h of stable transfection,the CCK-8 method was used to detect the light absorption(A)value of cells in each group at 24,48 and 72 h of culture,the scratch test was used to detect the fusion distance of cells in each group at 24 h of culture,the luciferase reporter gene assay was used to detect the relationship between LncRNA XIST,miR-193a-3p and RSF1,RT-qPCR was used to detect the expressions of lncRNA XIST,miR-193a-3p and RSF1 mRNA in each group,Western blot was used to detect the expression of RSF1 protein.Results The expression level of lncRNA XIST in osteosarcoma tissues was significantly higher than that in paracancerous tissues,and the difference was statistically significant(P<0.05).The A value in the blank group,no-load group and test group was increased with the transfection time extension,and the difference was statistically significant(P<0.05).Compared with the blank group and no-load group,the A value after 24,48,72 h of culture in the test group was decreased,the fusion distance was shortened,the expression of lncRNA XIST and RSF1 mRNA was decreased,the expression of miR-193a-3p mRNA was increased,the RSF1 protein was decreased,and differences were statistically significant(P<0.05).Co

关 键 词：骨肉瘤细胞 长链非编码RNA 微小RNA-193a-3p 增殖 迁移 

分 类 号：R738.1[医药卫生—肿瘤]