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环状 RNA 0067934 通过调控 miR-1182/KLF8 轴和激活 Wnt/β-连环蛋白通路促进非小细胞肺癌的发展。

Circ_0067934 promotes non-small cell lung cancer development by regulating miR-1182/KLF8 axis and activating Wnt/β-catenin pathway.

机构信息

Department of Respiratory and Critical Medicine, The Huaihe Hospital of Henan University, Kaifeng, Henan, China.

Department of Respiratory and Critical Medicine, The Huaihe Hospital of Henan University, Kaifeng, Henan, China.

出版信息

Biomed Pharmacother. 2020 Sep;129:110461. doi: 10.1016/j.biopha.2020.110461. Epub 2020 Jul 16.

Abstract

Non-small cell lung cancer (NSCLC) is the primary subtype of lung cancer with high mortality. Circular RNAs (circRNAs) play a crucial role in tumor development and progression. This study aimed to explore the function of circ_0067934 in NSCLC progression and its molecular basis. The levels of circ_0067934, miR-1182 and kruppel like factor 8 (KLF8) were measured by quantitative real-time polymerase chain reaction or western blot assay. Cell viability was detected by Cell Counting Kit-8 (CCK-8) assay. Cell migration and invasion were assessed by transwell assay. Cell apoptosis was monitored by flow cytometry. The protein levels of epithelial-to-mesenchymal transition (EMT)-related markers and Wnt/β-catenin pathway-related proteins were examined by western blot. Dual-luciferase reporter assay, RNA Immunoprecipitation (RIP) assay or RNA pull-down assay was performed to verify the interaction among circ_0067934, miR-1182 and KLF8. Xenograft assay was used to detect tumor growth in vivo. We found that circ_0067934 and KLF8 were up-regulated, while miR-1182 was down-regulated in NSCLC tissues and cells. Circ_0067934 knockdown blocked proliferation, migration, invasion and EMT and induced apoptosis in NSCLC cells. Circ_0067934 regulated NSCLC progression by sponging miR-1182. MiR-1182 targeted KLF8 to hinder NSCLC development. In addition, depletion of circ_0067934 restrained tumor growth in vivo. In conclusion, Circ_0067934 acted as a competing endogenous RNA to facilitate NSCLC progression by regulating the miR-1182/KLF8 axis and activating Wnt/β-catenin pathway.

摘要

非小细胞肺癌(NSCLC)是具有高死亡率的主要肺癌亚型。环状 RNA(circRNA)在肿瘤发生和发展中起着关键作用。本研究旨在探讨 circ_0067934 在 NSCLC 进展中的作用及其分子基础。通过实时定量聚合酶链反应或 Western blot 检测 circ_0067934、miR-1182 和 Kruppel 样因子 8(KLF8)的水平。通过细胞计数试剂盒-8(CCK-8)测定细胞活力。通过 Transwell 测定法评估细胞迁移和侵袭。通过流式细胞术监测细胞凋亡。通过 Western blot 检测上皮间质转化(EMT)相关标记物和 Wnt/β-catenin 通路相关蛋白的蛋白水平。通过双荧光素酶报告基因检测、RNA 免疫沉淀(RIP)测定或 RNA 下拉测定验证 circ_0067934、miR-1182 和 KLF8 之间的相互作用。通过异种移植实验检测体内肿瘤生长。我们发现,circ_0067934 和 KLF8 在 NSCLC 组织和细胞中上调,而 miR-1182 下调。circ_0067934 敲低可阻断 NSCLC 细胞的增殖、迁移、侵袭和 EMT,并诱导细胞凋亡。circ_0067934 通过海绵吸附 miR-1182 调节 NSCLC 进展。miR-1182 靶向 KLF8 以抑制 NSCLC 的发展。此外,circ_0067934 的耗竭抑制了体内肿瘤的生长。总之,circ_0067934 作为竞争性内源性 RNA,通过调节 miR-1182/KLF8 轴并激活 Wnt/β-catenin 通路,促进 NSCLC 的进展。

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