miRNA-200a-3p 通过靶向 IRS2 抑制非小细胞肺癌的增殖、迁移和侵袭。
MiRNA-200a-3p suppresses the proliferation, migration and invasion of non-small cell lung cancer through targeting IRS2.
机构信息
Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Jan;24(2):712-720. doi: 10.26355/eurrev_202001_20050.
OBJECTIVE
To uncover the biological role of microRNA-200a-3p (miRNA-200a-3p) in the progression of non-small cell lung cancer (NSCLC) and the underlying mechanism.
PATIENTS AND METHODS
The expression levels of miRNA-200a-3p and IRS2 in NSCLC tissues and cell lines were examined through quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between the miRNA-200a-3p level and pathological characteristics of NSCLC patients was analyzed. The prognostic value of miRNA-200a-3p in NSCLC was assessed through the Kaplan-Meier method. The potential interaction between miRNA-200a-3p and IRS2 was explored through Dual-Luciferase Reporter Gene Assay and Spearman correlation test. The regulatory effects of miRNA-200a-3p/IRS2 on the proliferative, migratory, and invasive abilities of NSCLC were evaluated by Cell Counting Kit-8 (CCK-8) and the transwell assay. The protein levels of the epithelial-mesenchymal transition (EMT)-related genes in NSCLC cells influenced by miRNA-200a-3p were detected by Western blot.
RESULTS
MiRNA-200a-3p was downregulated in NSCLC tissues and cell lines. The expression level of miRNA-200a-3p was related to tumor size, TNM staging, and lymphatic metastasis of NSCLC. The low level of miRNA-200a-3p predicted worse prognosis in NSCLC patients. The overexpression of miRNA-200a-3p inhibited A549 cells from proliferating, migrating, and invading. The protein levels of E-cadherin were upregulated, while N-cadherin and Vimentin were downregulated in A549 cells overexpressing miRNA-200a-3p. The Dual-Luciferase Reporter Gene Assay verified the binding between miRNA-200a-3p and IRS2. The level of IRS2 was negatively regulated by miRNA-200a-3p. Moreover, the overexpression of IRS2 could reverse the regulatory role of miRNA-200a-3p in the cellular behaviors of A549 cells.
CONCLUSIONS
MiRNA-200a-3p suppresses the proliferative, migratory, and invasive abilities of NSCLC by targeting IRS2, thus alleviating the progression of NSCLC.
目的
揭示微小 RNA-200a-3p（miRNA-200a-3p）在非小细胞肺癌（NSCLC）进展中的生物学作用及其潜在机制。
患者与方法
采用实时定量聚合酶链反应（qRT-PCR）检测 NSCLC 组织和细胞系中 miRNA-200a-3p 和 IRS2 的表达水平。分析 miRNA-200a-3p 水平与 NSCLC 患者病理特征的相关性。采用 Kaplan-Meier 法评估 miRNA-200a-3p 在 NSCLC 中的预后价值。通过双荧光素酶报告基因检测和 Spearman 相关分析探讨 miRNA-200a-3p 与 IRS2 之间的潜在相互作用。采用细胞计数试剂盒-8（CCK-8）和 Transwell 实验评估 miRNA-200a-3p/IRS2 对 NSCLC 增殖、迁移和侵袭能力的调控作用。采用 Western blot 检测 miRNA-200a-3p 对 NSCLC 细胞上皮间质转化（EMT）相关基因蛋白水平的影响。
结果
miRNA-200a-3p 在 NSCLC 组织和细胞系中表达下调。miRNA-200a-3p 的表达水平与 NSCLC 的肿瘤大小、TNM 分期和淋巴转移有关。miRNA-200a-3p 低表达预示 NSCLC 患者预后不良。过表达 miRNA-200a-3p 可抑制 A549 细胞增殖、迁移和侵袭。过表达 miRNA-200a-3p 可上调 A549 细胞中 E-钙黏蛋白的表达，下调 N-钙黏蛋白和波形蛋白的表达。双荧光素酶报告基因检测证实了 miRNA-200a-3p 与 IRS2 之间的结合。IRS2 的水平受 miRNA-200a-3p 的负调控。此外，过表达 IRS2 可逆转 miRNA-200a-3p 对 A549 细胞细胞行为的调节作用。
结论
miRNA-200a-3p 通过靶向 IRS2 抑制 NSCLC 的增殖、迁移和侵袭能力，从而缓解 NSCLC 的进展。
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