摘要
目的探讨长链非编码RNA尿路上皮癌相关基因1(Long non-coding RNA urothelial carcinoma related gene 1,LncRNA UCA1)对微小核糖核酸206(MicroRNA-206,miR-206)和CLOCk的表达影响及其在神经胶质瘤细胞增殖和细胞周期中的作用。方法采用实时荧光定量核酸扩增法(Real-time Quantitative PCR,qRT-PCR)检测在U251中与LncRNA UCA1和miR-206共转染细胞中IgG和Ago2的mRNA表达。分析生物学信息以预测LncRNA UCA1与miR-206之间的联系。采用跨孔转移实验和侵袭实验观察细胞的侵袭能力。采用蛋白质印迹法和免疫荧光法检测相关蛋白质的表达。最后通过动物实验了解LncRNA UCA1在神经胶质瘤细胞生长中的作用。结果LncRNA UCA1在神经胶质瘤细胞株和组织中表达上调,其通过诱导上皮-间质转化,提高了胶质瘤细胞的侵袭能力。LncRNA UCA1能够调节miR-206的表达,并作为miR-206的内源因子。EMT诱导剂CLOCk是miR-206的信使RNA靶点。LncRNA UCA1在体内通过靶向调节miR-206和CLOCk从而发挥生物学功能。结论LncRNA UCA1/miR-206/CLOCk轴能够调节神经胶质瘤细胞的增殖和侵袭,将来可能成为神经胶质瘤的一个潜在治疗靶点。
Objective This study will discuss the expression of LncRNA UCA1 regarding miR-206 and CLOCK,and their integrative efects in the proliferation and cell cycle of glioma cells.Methods qRT-PCR was conducted to measure the mRNA expressions of IgG and Ago2 in cells co-transfected with LncRNA UCA1,and miR-206 in U251.Bioinformation was analyzed for the prediction of association between LncRNA UCA1 and miR-206.Transwell migrations assays and invasion assays were utilized to observe the cell invasive ability.Western blot and immunofuorescence imaging were used to examine the protein expressions.In vivo comparisons and observations were also performed to investigate the role of LncRNA UCA1 in glioma growth.Results LncRNA UCA1 was up-regulated in glioma cell lines and tissues.It elevated cell invasion via the inducing of epithelial-mesenchymal transition.We found that LncRNA UCA1 can modulate miR-206 expression and serve as an endogenous sponge of miR-206.The EMT-inducer CLOCK was validated as a messenger RNA target of miR-206.At last,we demonstrated that LncRNA UCA1 exerted the biology function through regulating miR-206 and CLOCK in vivo.Conclusions LncRNA UCA1/miR-206/CLOCk axis can regulate the proliferation and invasion of glioma cells and may become a potential therapeutic target for gliomas in the future.
作者
吕洋
黄智
向雷
蒋天鹏
李兴
Lv Yang;Huang Zhi;Xiang Lei;Jiang Tianpeng;Li Xing(School of Medical Imaging,Guizhou Medical University,Guiyang 550004,Guizhou,China)
出处
《贵州医药》
CAS
2020年第12期1851-1854,F0003,共5页
Guizhou Medical Journal
基金
贵州省普通高等学校医学影像工程研究中心[黔教合KY字(2016)012]
贵州医科大学医学影像工程研究中心(核工程中心2016001)。
