摘要
目的:探讨及分析骨肉瘤中差异表达长链非编码RNA(long non-coding RNA,lncRNA)的表达谱及功能。方法:本课题前期通过5对组织(骨肉瘤组织和瘤旁组织)进行高通量转录组学测序筛选出差异表达的lncRNAs,并筛选出上调变化的前5位lncRNAs,通过荧光定量PCR在骨肉瘤细胞系内检测lncRNA MAFG-AS1的差异表达,并通过小干扰RNA技术干扰其表达,CCK-8法检测敲低lncRNA后细胞增殖是否发生变化,Western-blotting检测下游蛋白变化。结果:高通量数据分词浅析显示,共有376个lncRNAs在骨肉瘤组织中差异表达,其中206个上调,170个下调,并筛选出上调变化的前5位lncRNAs。荧光定量PCR显示lncRNA MAFG-AS1在骨肉瘤细胞内高表达。CCK-8法结果显示该lncRNA MAFG-AS1具有影响骨肉瘤细胞增殖的作用,且下游蛋白RRM2表达发生变化。结论:骨肉瘤组织中存在明显的lncRNAs差异性表达。且lncRNA MAFG-AS1在骨肉瘤发生和发展中发挥中重要的作用。
Objective： To explore and analyze the expression profile and analysis function of differentially expressed long non-coding RNA（ lncRNA） in osteosarcoma. Methods： In this study,5 pairs of tissues（ osteosarcoma and normal tissues） were screened for high-throughput transcriptome sequencing to screen for differentially expressed lncRNAs. The top five lncRNAs were screened for up-regulation and quantified by fluorescence quantitative PCR. The differential expression of lncRNA MAFG-AS1 was detected in the cell lines,and the expression of lncRNA MAFG-AS1 was interfered by small interfering RNA technology. The CCK-8 assay was used to detect cell viability.Western-blotting was used to detect downstream protein changes. Results： High-throughput data analysis showed that a total of 376 lncRNAs were differentially expressed in osteosarcoma tissue,of which 206 were up-regulated and170 were down-regulated,and the top 5 lncRNAs were selected for up-regulation change. Fluorescent quantitative PCR showed lncRNA MAFG-AS1 in osteosarcoma showed high expression in cells. CCK-8 results showed that the lncRNA MAFG-AS1 had an effect on the proliferation of osteosarcoma cells,and changes in downstream protein levels. Conclusion： There is a distinct expression of lncRNAs in osteosarcoma tissues. And lncRNA MAFG-AS1 plays an important role in the occurrence and development of osteosarcoma.
作者
张浩萌
马琼
杨童磊
张甜
周勇
Zhang Haomeng;Ma Qiong;Yang Tonglei;Zhang Tian;Zhou Yong(Department of Orthopedics;Department of Pharmacy,Tangdu Hospital,Air Force Medical University,Shaanxi Xi＇an 710038,China)
出处
《现代肿瘤医学》
CAS
2018年第22期3536-3540,共5页
Journal of Modern Oncology
基金
国家自然科学基金资助项目(编号:81272441)
