Zhou Fu-Chao, Zhang Yue-Hui, Liu Hai-Tao, Song Jia, Shao Jiang
Spine Center, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Front Pharmacol. 2020 Mar 24;11:255. doi: 10.3389/fphar.2020.00255. eCollection 2020.
Long non-coding RNAs (lncRNAs) are being found to play an increasingly important role in the development of tumors. However, their biological functions and the underlying mechanisms remain unclear. Using information from GEO Datasets, we found that the lncRNA LINC00588 was downregulated in osteosarcoma (OS) in bone but was upregulated in the metastatic tumor present in the lung. We assessed the function of LINC00588 using both overexpression and knock-out studies. We performed colony formation assay, CCK-8 assay, flow cytometry, wound healing assay, transwell assay, and RT-qPCR assay and used a xenograft model to investigate the influence of LINC00588 on cell proliferation, viability, cell apoptosis and cycle, migration, invasion, endothelial cell function, EMT (epithelial to mesenchymal transition), and tumor growth, respectively. Overexpression of LINC00588 appeared to inhibit cell proliferation, viability, migration, invasion, endothelial cell function, EMT, and tumor growth but not apoptosis, while we got the opposite result when we knocked down LINC00588. Next, we predicted that LINC00588 bound to miRNA-1972 and significantly downregulated its expression, which we then verified through a luciferase reporter assay. Subsequently, we knocked down miR1972 and performed CCK-8 and transwell assays to demonstrate that downregulation of miRNA-1972 could substantially inhibit the viability and invasion of osteosarcoma cells. The expression of TP53 was downregulated at the protein level but not at the mRNA level after the overexpression of miRNA-1972. Taken together, our findings indicate that LINC00588 plays a role in OS development by downregulating the expression of miRNA-1972, which can, in turn, inhibit the expression of TP53. Hence, we believe that the LINC00588/miRNA-1072/TP53 axis could potentially serve as a therapeutic target or diagnostic biomarker for osteosarcoma.
长链非编码RNA(lncRNAs)在肿瘤发生发展过程中发挥着越来越重要的作用。然而,其生物学功能及潜在机制仍不清楚。利用来自基因表达综合数据库(GEO Datasets)的信息,我们发现lncRNA LINC00588在骨肉瘤(OS)的原发瘤中表达下调,但在肺转移瘤中表达上调。我们通过过表达和敲除实验评估了LINC00588的功能。我们进行了集落形成实验、CCK-8实验、流式细胞术、伤口愈合实验、Transwell实验和逆转录定量PCR实验,并利用异种移植模型分别研究了LINC00588对细胞增殖、活力、细胞凋亡和周期、迁移、侵袭、内皮细胞功能、上皮-间质转化(EMT)以及肿瘤生长的影响。LINC00588的过表达似乎抑制了细胞增殖、活力、迁移、侵袭、内皮细胞功能、EMT以及肿瘤生长,但对细胞凋亡没有影响,而敲低LINC00588时我们得到了相反的结果。接下来,我们预测LINC00588与miRNA-1972结合并显著下调其表达,随后通过荧光素酶报告基因实验进行了验证。随后,我们敲低miR1972并进行CCK-8和Transwell实验,以证明miRNA-1972的下调可显著抑制骨肉瘤细胞的活力和侵袭。miRNA-1972过表达后,TP53的蛋白水平下调,但mRNA水平未下调。综上所述,我们的研究结果表明,LINC00588通过下调miRNA-1972的表达在骨肉瘤发生发展中发挥作用,而miRNA-1972又可抑制TP53的表达。因此,我们认为LINC00588/miRNA-1072/TP53轴可能作为骨肉瘤的治疗靶点或诊断生物标志物。
原文中“miRNA-1072”应为“miRNA-1972”,译文已修正。