长链非编码RNA LINC01419介导miR-519a-3p/PDRG1轴促进骨肉瘤细胞进展。
Long non-coding RNA LINC01419 mediates miR-519a-3p/PDRG1 axis to promote cell progression in osteosarcoma.
作者信息
Gu Zhiqian, Wu Shaokun, Wang Jingnan, Zhao Shoujun
机构信息
Department of Orthopedics, Hwa Mei Hospital, University of Chinese Academy of Sciences (Ningbo NO. 2 Hospital), No. 41 Northwest Street, Ningbo, 315010 Zhejiang China.
Ningbo Institute of Life and Health Industry, No. 41 Northwest Street, Ningbo, 315010 Zhejiang China.
出版信息
Cancer Cell Int. 2020 May 5;20:147. doi: 10.1186/s12935-020-01203-0. eCollection 2020.
BACKGROUND
Osteosarcoma (OS) is one of the most aggressive malignancies with mortality rate worldwide. Accumulating evidence has revealed that long noncoding RNAs (lncRNAs) exert important functions in regulation of cancer initiation and progression. Recently, long intergenic non-protein coding RNA 1419 (LINC01419) has been reported to function as an oncogene in several cancers. However, its role in OS has not been explored yet.
METHODS
qRT-PCR and western blot analyses were implemented to determine the expression of genes. The function of OS cells was assessed through colony formation, EdU, JC-1, TUNEL, transwell, and immunofluorescence (IF) assays. FISH and subcellular fractionation assays were conducted to estimate the localization of LINC01419 in OS cells. The interaction between genes was validated through luciferase reporter and RNA pull down assays.
RESULTS
LINC01419 expression was elevated in OS tissues and cells. Functionally, LINC01419 accelerated OS cell proliferation, motility and EMT. In vivo assay showed that silencing LINC01419 hindered the growth of OS tumors. Mechanistic investigation unveiled that LINC01419 acted as a competing endogenous RNA (ceRNA) to augment PDRG1 expression by miR-519a-3p sequestration. Rescue assays verified the oncogenic effect of LINC01419/miR-519a-3p/PDRG1 axis on OS development.
CONCLUSION
LINC01419 mediates malignant phenotypes in OS by targeting miR-519a-3p/PDRG1 axis.
背景
骨肉瘤（OS）是全球死亡率最高的侵袭性最强的恶性肿瘤之一。越来越多的证据表明，长链非编码RNA（lncRNA）在癌症的发生和发展调控中发挥着重要作用。最近，有报道称长链基因间非编码RNA 1419（LINC01419）在几种癌症中作为癌基因发挥作用。然而，其在骨肉瘤中的作用尚未得到探索。
方法
采用qRT-PCR和蛋白质免疫印迹分析来确定基因的表达。通过集落形成、EdU、JC-1、TUNEL、transwell和免疫荧光（IF）试验评估骨肉瘤细胞的功能。进行荧光原位杂交（FISH）和亚细胞分级分离试验以估计LINC01419在骨肉瘤细胞中的定位。通过荧光素酶报告基因和RNA下拉试验验证基因之间的相互作用。
结果
LINC01419在骨肉瘤组织和细胞中表达升高。在功能上，LINC01419促进了骨肉瘤细胞的增殖、迁移和上皮-间质转化（EMT）。体内试验表明，沉默LINC01419可抑制骨肉瘤肿瘤的生长。机制研究表明，LINC01419作为一种竞争性内源性RNA（ceRNA），通过隔离miR-519a-3p来增加PDRG1的表达。挽救试验证实了LINC01419/miR-519a-3p/PDRG1轴对骨肉瘤发展的致癌作用。
结论
LINC01419通过靶向miR-519a-3p/PDRG1轴介导骨肉瘤的恶性表型。
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