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Circ_0072083 interference enhances growth-inhibiting effects of cisplatin in non-small-cell lung cancer cells via miR-545-3p/CBLL1 axis
Cancer Cell International ( IF 5.3 ) Pub Date : 2020-03-12 , DOI: 10.1186/s12935-020-1162-x
Huiping Li 1 , Fangfang Liu 1 , Wenjing Qin 1
Affiliation  

Non-small-cell lung cancer (NSCLC) is one of the common cancers in the world. Circular RNA 0072083 (circ_0072083, circZFR) has been reported to be associated with the progression of NSCLC. In this study, we intended to explore the role and the potential mechanism of circ_0072083 in NSCLC. Quantitative real time polymerase chain reaction (qRT-PCR) was performed to detect the expression of circ_0072083, its matching linear RNA (zinc finger RNA binding protein (ZFR)) and microRNA-545-3p (miR-545-3p) in NSCLC cells. The ability of colony formation in NSCLC cells was detected by colony formation assay. The apoptosis and cell cycle were measured by flow cytometry. The metastasis was determined by transwell migration and invasion assays. The protein expression of E-cadherin, N-cadherin, Vimentin and Cbl proto-oncogene like 1 (CBLL1) was examined by western blot assay. The interaction between miR-545-3p and circ_0072083 or CBLL1 was predicted by starBase or Targetscan software. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were applied to validate these interactions. Nude mice bearing tumors were used to confirm the role of circ_0072083 and cisplatin (DDP) in vivo. The level of circ_0072083 was higher in NSCLC tissues and cells relative to that in adjacent non-tumor tissues and normal lung cells. The transfection of si-circ_0072083 inhibited colony formation, cell cycle and metastasis while promoted the apoptosis of NSCLC cells stimulated by DDP. MiR-545-3p was a direct functional target of circ_0072083 in NSCLC cells. CBLL1 could bind to miR-545-3p in NSCLC cells. Circ_0072083 promoted the progression of NSCLC induced by DDP through sponging miR-545-3p and enhancing the enrichment of CBLL1 in vivo and in vitro. Circ_0072083 depletion contributed to DDP-triggered inhibition of NSCLC tumor through miR-545-3p/CBLL1 axis.

中文翻译:

Circ_0072083 干扰通过 miR-545-3p/CBLL1 轴增强顺铂对非小细胞肺癌细胞的生长抑制作用

非小细胞肺癌(NSCLC)是世界上常见的癌症之一。据报道,环状 RNA 0072083 (circ_0072083, circZFR) 与 NSCLC 的进展有关。本研究旨在探讨circ_0072083在NSCLC中的作用及其潜在机制。进行定量实时聚合酶链反应 (qRT-PCR) 以检测 circ_0072083 及其匹配的线性 RNA(锌指 RNA 结合蛋白 (ZFR))和 microRNA-545-3p (miR-545-3p) 在 NSCLC 细胞中的表达. 通过集落形成试验检测NSCLC细胞中集落形成的能力。流式细胞仪检测细胞凋亡和细胞周期。通过transwell迁移和侵袭测定确定转移。E-钙粘蛋白、N-钙粘蛋白的蛋白表达,通过蛋白质印迹测定检查波形蛋白和Cbl原癌基因样1(CBLL1)。通过starBase或Targetscan软件预测miR-545-3p与circ_0072083或CBLL1的相互作用。应用双荧光素酶报告基因测定和 RNA 免疫沉淀 (RIP) 测定来验证这些相互作用。使用带有肿瘤的裸鼠来确认 circ_0072083 和顺铂 (DDP) 在体内的作用。相对于邻近的非肿瘤组织和正常肺细胞,circ_0072083 在 NSCLC 组织和细胞中的水平更高。si-circ_0072083转染抑制集落形成、细胞周期和转移,同时促进DDP刺激的NSCLC细胞凋亡。MiR-545-3p 是 circ_0072083 在 NSCLC 细胞中的直接功能靶点。CBLL1 可以与 NSCLC 细胞中的 miR-545-3p 结合。Circ_0072083 通过海绵化 miR-545-3p 和增强 CBLL1 在体内和体外的富集来促进 DDP 诱导的 NSCLC 的进展。Circ_0072083 缺失有助于 DDP 通过 miR-545-3p/CBLL1 轴抑制 NSCLC 肿瘤。
更新日期:2020-03-12
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