长链非编码 RNA FENDRR 通过海绵吸附 miR-761 抑制 NSCLC 细胞生长和侵袭。
Long non-coding RNA FENDRR inhibits NSCLC cell growth and aggressiveness by sponging miR-761.
机构信息
Department of Thoracic Surgery, Affiliated Hospital of Weifang Medical University, Weifang, China.
出版信息
Eur Rev Med Pharmacol Sci. 2018 Dec;22(23):8324-8332. doi: 10.26355/eurrev_201812_16530.
OBJECTIVE
The aim of this paper is to investigate the functions of long noncoding RNA (lncRNA) FOXF1 Adjacent Non-Coding Developmental Regulatory RNA (FENDRR) in the growth and aggressiveness of non-small cell lung cancer (NSCLC).
PATIENTS AND METHODS
The expression of FENDRR in NSCLC tissues and cell lines was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and colony formation assays were conducted to explore the roles of FENDRR on the growth of NSCLC cell. The wound healing and transwell invasion assays were conducted to explore the impact of FENDRR on NSCLC cell migration and invasion. The apoptosis of NSCLC cell was detected using flow cytometer-based Annexin V/Propidium Iodide (PI) dual staining. The xenograft model was conducted to investigate the effect of FENDRR on the growth of NSCLC cell in vivo. The expression of Ki67 was measured by immunohistochemical (IHC) staining using Ki67 antibody. Bioinformatics analysis and Luciferase reporter assay were applied to identify that miR-761 was the target of FENDRR. Additional, colony formation and transwell experiments were utilized to confirm that FENDRR inhibited the growth and aggressiveness of NSCLC cell by regulating miR-761.
RESULTS
We found a marked down-regulation of FENDRR in NSCLC tissues compared to tumor-adjacent tissues. FENDRR down-expression was detected in four NSCLC cell lines (H1650, HCC827, H1975 and A549) compared to the human non-tumorigenic bronchial epithelial cell, BEAS-2B. Low expression of FENDRR was identified as a predictive factor for poor prognosis of patients with NSCLC. The over-regulation of FENDRR inhibited the proliferation, migration and invasion capacities of NSCLC cell and promoted the apoptosis of NSCLC cell in vitro whereas the down-regulation of FENDRR caused the opposite results. Moreover, the over-expression of FENDRR restrained the growth of NSCLC cell in vivo. We found that there were potential binding sites between FENDRR and miR-761 and the level of miR-761 was inversely associated with the expression of ENDRR in NSCLC tissues. Finally, the rescue experiments suggested that the anti-oncogenic role of FENDRR was at least partially mediated by miR-761 in NSCLC.
CONCLUSIONS
We found that FENDRR was down-expressed in NSCLC and the over-expression of FENDRR inhibited the malignant phenotypes of NSCLC cell by binding to miR-761 competitively.
目的
本文旨在研究长非编码 RNA（lncRNA）FOXF1 相邻非编码发育调控 RNA（FENDRR）在非小细胞肺癌（NSCLC）生长和侵袭中的作用。
患者和方法
采用实时荧光定量聚合酶链反应（qRT-PCR）检测 NSCLC 组织和细胞系中 FENDRR 的表达。通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）和集落形成实验探讨 FENDRR 对 NSCLC 细胞生长的作用。通过划痕愈合和 Transwell 侵袭实验探讨 FENDRR 对 NSCLC 细胞迁移和侵袭的影响。采用流式细胞术碘化丙啶（PI）双染法检测 NSCLC 细胞的凋亡。通过异种移植模型研究 FENDRR 对 NSCLC 细胞体内生长的影响。采用 Ki67 抗体进行免疫组织化学（IHC）染色检测 Ki67 的表达。应用生物信息学分析和荧光素酶报告基因实验鉴定 miR-761 是 FENDRR 的靶基因。另外，通过集落形成和 Transwell 实验证实 FENDRR 通过调节 miR-761 抑制 NSCLC 细胞的生长和侵袭能力。
结果
与肿瘤相邻组织相比，我们发现 NSCLC 组织中 FENDRR 的表达明显下调。与人类非肿瘤性支气管上皮细胞 BEAS-2B 相比，在四种 NSCLC 细胞系（H1650、HCC827、H1975 和 A549）中检测到 FENDRR 下调。FENDRR 低表达被鉴定为 NSCLC 患者预后不良的预测因素。FENDRR 的过表达抑制 NSCLC 细胞的增殖、迁移和侵袭能力，并促进 NSCLC 细胞的凋亡，而 FENDRR 的下调则导致相反的结果。此外，FENDRR 的过表达抑制了 NSCLC 细胞在体内的生长。我们发现 FENDRR 和 miR-761 之间存在潜在的结合位点，并且 NSCLC 组织中 miR-761 的水平与 FENDRR 的表达呈负相关。最后，通过 rescue 实验表明，FENDRR 通过与 miR-761 竞争结合发挥其抑制 NSCLC 细胞恶性表型的部分作用。
结论
我们发现 FENDRR 在 NSCLC 中表达下调，FENDRR 的过表达通过与 miR-761 竞争结合抑制 NSCLC 细胞的恶性表型。
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