长链非编码 RNA SNHG1 通过 miR-330-5p/DCLK1 轴促进 NSCLC 的顺铂耐药和进展。
LncRNA SNHG1 contributes to the cisplatin resistance and progression of NSCLC via miR-330-5p/DCLK1 axis.
机构信息
Shaanxi Provincial Research Center for the Project of Prevention and Treatment of Respiratory Diseases, Xi'an Medical University, Xi'an City, Shaanxi Province, China; Department of Cardiothoracic Surgery, The Second Affiliated Hospital of Xi'an Medical University, Xi'an City, Shaanxi Province, China.
Department of Gynecology, The Second Affiliated Hospital of Xi'an Medical University, Xi'an City, Shaanxi Province, China.
出版信息
Exp Mol Pathol. 2021 Jun;120:104633. doi: 10.1016/j.yexmp.2021.104633. Epub 2021 Mar 19.
BACKGROUND
Long non-coding RNAs (lncRNAs) are involved in the occurrence and progression of multiple cancers, including non-small cell lung cancer (NSCLC). Herein, we explored the exact role and underlying mechanism of lncRNA small nucleolar RNA host gene 1 (SNHG1) in NSCLC.
METHODS
The levels of SNHG1, microRNA-330-5p (miR-330-5p) and doublecortin-like kinase 1 (DCLK1) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was carried out to measure the chemoresistance and proliferation of NSCLC cells. The metastasis and apoptosis of NSCLC cells were examined by transwell migration and invasion assays and flow cytometry. Western blot assay was conducted to detect the levels of proliferation-associated proteins and DCLK1. The interaction between miR-330-5p and SNHG1 or DCLK1 was predicted by StarBase and microT_CDS databases. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to validate these interactions. In vivo chemosensitivity experiment was conducted to assess the function of SNHG1 in the chemoresistance of NSCLC in vivo.
RESULTS
SNHG1 was dramatically up-regulated in cisplatin (DDP)-resistant NSCLC tissues and cells. SNHG1 promoted the DDP resistance and malignant behaviors of NSCLC cells. SNHG1 functioned through targeting miR-330-5p, and si-SNHG1-mediated effects in NSCLC cells were attenuated by the addition of in-miR-330-5p. DCLK1 messenger RNA (mRNA) could directly bind to miR-330-5p, and miR-330-5p acted as a tumor suppressor in NSCLC through down-regulating DCLK1. SNHG1 silencing elevated the DDP sensitivity of NSCLC cells in vivo.
CONCLUSION
SNHG1 elevated DDP resistance and malignant potential of NSCLC cells through elevating the level of DCLK1 via sponging miR-330-5p.
背景
长链非编码 RNA（lncRNA）参与多种癌症的发生和发展，包括非小细胞肺癌（NSCLC）。在此，我们探讨了 lncRNA 小核仁 RNA 宿主基因 1（SNHG1）在 NSCLC 中的确切作用和潜在机制。
方法
通过实时定量聚合酶链反应（qRT-PCR）检测 SNHG1、微小 RNA-330-5p（miR-330-5p）和双皮质激酶 1（DCLK1）的水平。3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）试验用于测量 NSCLC 细胞的化疗耐药性和增殖。通过 Transwell 迁移和侵袭试验和流式细胞术检测 NSCLC 细胞的转移和凋亡。Western blot 试验检测增殖相关蛋白和 DCLK1 的水平。通过 StarBase 和 microT_CDS 数据库预测 miR-330-5p 与 SNHG1 或 DCLK1 之间的相互作用。双荧光素酶报告基因试验和 RNA 免疫沉淀（RIP）试验验证这些相互作用。进行体内化疗敏感性实验以评估 SNHG1 在体内 NSCLC 化疗耐药中的作用。
结果
SNHG1 在顺铂（DDP）耐药 NSCLC 组织和细胞中显著上调。SNHG1 促进了 NSCLC 细胞的 DDP 耐药性和恶性行为。SNHG1 通过靶向 miR-330-5p 发挥作用，并且在 NSCLC 细胞中添加 in-miR-330-5p 可减弱 si-SNHG1 介导的作用。DCLK1 信使 RNA（mRNA）可直接与 miR-330-5p 结合，miR-330-5p 通过下调 DCLK1 在 NSCLC 中发挥肿瘤抑制作用。SNHG1 沉默可提高 NSCLC 细胞在体内的 DDP 敏感性。
结论
SNHG1 通过海绵吸附 miR-330-5p 升高 DCLK1 水平，从而提高 NSCLC 细胞的 DDP 耐药性和恶性潜能。
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