miR-138-5p reverses gefitinib resistance in non-small cell lung cancer cells via negatively regulating G protein-coupled receptor 124
miR-138-5p reverses gefitinib resistance in non-small cell lung cancer cells via negatively regulating G protein-coupled receptor 124
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miR-138-5p 通过负调节 G 蛋白偶联受体 124 逆转非小细胞肺癌细胞对吉非替尼的耐药性
DOI：
10.1016/j.bbrc.2014.02.073
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发表时间：
2014-03-28
影响因子：
3.1
通讯作者：
Fu, XiangNing
中科院分区：
文献类型：
--
作者：
Gao, Yi;Fan, XiaoWu;Fu, XiangNing
表皮生长因子受体酪氨酸激酶抑制剂（EGFR TKI）如吉非替尼是EGFR激活突变非小细胞肺癌（NSCLC）患者的临床有效治疗药物。然而，治疗效果最终受限于获得性TKI耐药性的发展。microRNA（miRNAs）是一类在人类恶性肿瘤中普遍失调的小分子非编码RNA。本研究的目的是研究miRNAs在吉非替尼耐药中的作用。我们通过将PC 9 NSCLC细胞连续暴露于吉非替尼6个月，建立了吉非替尼耐药细胞模型（PC 9 GR）。miRNA微阵列筛选显示miR-138- 5 p在PC 9 GR细胞中表现出最大的下调。miR-138- 5 p的再表达足以使PC 9 GR细胞和另一种吉非替尼耐药NSCLC细胞系H1975对吉非替尼敏感。生物信息学分析和荧光素酶报告基因分析表明，G蛋白偶联受体124（GPR 124）是miR-138- 5 p的直接靶点。实验验证表明，miR-138- 5 p在NSCLC细胞中在蛋白和mRNA水平上抑制GPR 124的表达。此外，我们观察到肺腺癌标本中miR-138- 5 p和GPR 124的表达之间呈负相关。GPR 124的敲低模拟了miR-138- 5 p对吉非替尼敏感性的影响。总之，我们的研究结果表明，下调miR-138- 5 p有助于吉非替尼耐药，恢复miR-138- 5 p或抑制GPR 124可能是克服NSCLC吉非替尼耐药的潜在治疗方法。(c)2014爱思唯尔公司All rights reserved.
Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR TKIs) such as gefitinib are clinically effective treatments for non-small cell lung cancer (NSCLC) patients with EGFR activating mutations. However, therapeutic effect is ultimately limited by the development of acquired TKI resistance. MicroRNAs (miRNAs) represent a category of small non-coding RNAs commonly deregulated in human malignancies. The aim of this study was to investigate the role of miRNAs in gefitinib resistance. We established a gefitinib-resistant cell model (PC9GR) by continually exposing PC9 NSCLC cells to gefitinib for 6 months. MiRNA microarray screening revealed miR-138-5p showed the greatest downregulation in PC9GR cells. Re-expression of miR-138-5p was sufficient to sensitize PC9GR cells and another gefitinibresistant NSCLC cell line, H1975, to gefitinib. Bioinformatics analysis and luciferase reporter assay showed that G protein-coupled receptor124 (GPR124) was a direct target of miR-138-5p. Experimental validation demonstrated that expression of GPR124 was suppressed by miR-138-5p on protein and mRNA levels in NSCLC cells. Furthermore, we observed an inverse correlation between the expression of miR-138-5p and GPR124 in lung adenocarcinoma specimens. Knockdown of GPR124 mimicked the effects of miR-138-5p on the sensitivity to gefitinib. Collectively, our results suggest that downregulation of miR-138-5p contributes to gefitinib resistance and that restoration of miR-138-5p or inhibition GPR124 might serve as potential therapeutic approach for overcoming NSCLC gefitinib resistance. (c) 2014 Elsevier Inc. All rights reserved.