背景:长基因间非蛋白质编码RNA173(LINC00173)在各种癌症中执行重要功能。然而,其在鼻咽癌(NPC)中的作用和表达尚待研究。这里,我们研究了其对NPC恶性特征的影响,并阐明了LINC00173在NPC进展中的潜在分子机制。
方法:进行定量实时逆转录PCR(qRT-PCR)和免疫印迹以评估LINC00173,microRNA-765(miR-765),和Gremlin1(GREM1)在鼻咽癌细胞和组织中的表达。细胞计数试剂盒-8(CCK8),菌落形成,并进行了伤口愈合实验来评估增殖,增长,和NPC细胞的迁移,分别。通过异种移植肿瘤实验评估体内NPC细胞的肿瘤生长。此外,通过生物信息学分析研究了miR-765、LINC00173和GREM1之间的相互作用,荧光素酶报告基因和RNA免疫沉淀芯片测定。
结果:在NPC细胞系和组织中发现LINC00173表达上调。功能实验发现其下调抑制了NPC细胞的增殖,增长,和移民。此外,LINC00173敲除阻碍了体内NPC细胞的肿瘤生长。通过下调miR-765可以部分逆转这些作用。GREM1是miR-765的下游靶标。GREM1击倒可以抑制扩散,增长,和NPC细胞的迁移。尽管如此,miR-765下调可以消除这些抗肿瘤作用.机械上,LINC00173通过与miR-765结合增加GREM1的表达。
结论:LINC00173通过与miR-765结合,通过GREM1上调促进鼻咽癌进展,从而发挥致癌因子的作用。这项研究为NPC进展中涉及的分子机制提供了新的见解。
BACKGROUND: Long intergenic non-protein-coding RNA 173 (LINC00173) executes vital functions in various cancers. Nevertheless, its role and expression in nasopharyngeal carcinoma (NPC) have yet to be investigated. Here, we investigated its effects on the malignancy characteristics of NPC and elucidated the potential molecular mechanism of LINC00173 in NPC progression.
METHODS: Quantitative real-time reverse transcription-PCR (qRT-PCR) and immunoblotting were conducted to estimate the LINC00173, microRNA-765 (miR-765), and Gremlin 1 (GREM1) expressions in NPC cells and tissues. Cell counting kit-8 (CCK8), colony formation, and wound healing experiments were done to evaluate the proliferation, growth, and migration of NPC cells, respectively. The tumorous growth of NPC cells in vivo was assessed through the xenograft tumor experiment. Furthermore, the interactions among miR-765, LINC00173, and GREM1 were investigated through bioinformatics analyses, luciferase reporter and RNA immunoprecipitation chip assays.
RESULTS: An upregulated LINC00173 expression was found in NPC cell lines and tissues. The functional experiments uncovered that its downregulation repressed NPC cell proliferation, growth, and migration. In addition, LINC00173 knockdown hampered the NPC cells\' tumorous growth in vivo. These effects could partially be reversed by downregulating miR-765. GREM1 is a downstream target of miR-765. GREM1 knockdown could repress the proliferation, growth, and migration of NPC cells. Nonetheless, these anti-tumor effects could be abolished by miR-765 downregulation. Mechanistically, LINC00173 increased the expression of GREM1 by binding with miR-765.
CONCLUSIONS: LINC00173 functions as an oncogenic factor by binding with miR-765 to promote the progression of NPC via GREM1 upregulation. This study provides a novel insight into the molecular mechanisms involved in NPC progression.