MiR-21 overexpression is associated with acquired resistance of EGFR-TKI in non-small cell lung cancer
MiR-21 overexpression is associated with acquired resistance of EGFR-TKI in non-small cell lung cancer
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MiR-21 过表达与非小细胞肺癌中 EGFR-TKI 的获得性耐药相关。
DOI：
10.1016/j.lungcan.2013.11.003
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发表时间：
2014-02-01
期刊：
影响因子：
5.3
通讯作者：
Schmit-Bindert, Gerald
中科院分区：
文献类型：
--
作者：
Li, Bing;Ren, Shengxiang;Schmit-Bindert, Gerald
背景与目的：随着表皮生长因子受体酪氨酸激酶抑制剂（EGFR TKI）在晚期非小细胞肺癌（NSCLC）患者中的应用越来越多，其获得性耐药已成为一个重大的临床问题。最近的研究表明，miR-21参与了细胞毒性药物的耐药性。本研究的目的是探讨其在NSCLC对EGFR-TKI获得性耐药中的作用。方法：采用egfr - tki敏感的人肺腺癌细胞株PC9和获得性耐药细胞株PC9R。采用慢病毒载体感染PC9或PC9R，调节miR-21的表达。靶蛋白PTEN和PDCD4的表达受RNA干扰控制。采用MicroRNA阵列、RT-PCR和TaqMan MicroRNA检测miR-21的表达。MTT和Annexin V检测细胞增殖和凋亡。Western Blot和免疫组化分析靶蛋白（PTEN、PDCD4、Akt、p-Akt）表达。我们还构建了PC9R异种移植肿瘤模型，在体内观察miR-21与EGFR-TKI耐药之间的关系，并在EGFR-TKI治疗的NSCLC患者的临床血清标本中进行验证。结果：相对于PC9， MiR-21在EGFR-TKI耐药细胞系PC9R中过表达。miR-21水平与PTEN、PDCD4表达呈负相关，与PI3K/Akt通路呈正相关。慢病毒载体抑制miR-21诱导PC9R细胞系凋亡，ASO抑制miR-21抑制EGFR-TKI裸鼠肿瘤生长。此外，接受EGFR-TKI治疗的NSCLC患者在获得耐药时血清miR-21表达显著高于基线（p < 0.01）。结论：miR-21通过下调PTEN和PDCD4，激活PI3K/Akt通路介导EGFR-TKI在NSCLC中获得性耐药。2013爱思唯尔爱尔兰有限公司版权所有。
Background and purpose: With the increasing use of epidermal growth factor receptor tyrosine kinase inhibitor (EGFR TKI) in patients with advanced non-small cell lung cancer (NSCLC), its acquired resistance has become a major clinical problem. Recent studies revealed that miR-21 was involved into the resistance of cytotoxic agents. The aim of this study was to investigate its role in the acquired resistance of NSCLC to EGFR-TKI.Methods: EGFR-TKI-sensitive human lung adenocarcinoma cell line PC9 and the acquired resistant cell line, PC9R, were used. Lentiviral vectors were used to infect PC9 or PC9R to regulate the miR-21 expression. The expression of targeted proteins PTEN and PDCD4 was controlled by RNA interference. MicroRNA array, RT-PCR and TaqMan MicroRNA Assays were used to detect miR-21 expression. The MTT and Annexin V assays were used to determine proliferation and apoptosis. Western Blot and immunohistochemistry were used to analyze target protein expression (PTEN, PDCD4, Akt, p-Akt). We also constructed PC9R xenograft tumor model to observe the relationship between miR-21 and EGFR-TKI resistance in vivo and validated it in the clinical serum specimens of NSCLC patients treated with EGFR-TKI.Result: MiR-21 was overexpressed in the EGFR-TKI resistant cell line PC9R relative to PC9. The level of miR-21 was reversely correlated with the expression of PTEN and PDCD4 and positive correlated with PI3K/Akt pathway. Inhibiting miR-21 with lentivirus vector induces apoptosis in PC9R cell line and inhibiting miR-21with ASO suppressed tumor growth in nude mice treated with EGFR-TKI. Furthermore, serum miR-21 expression in NSCLC patients treated with EGFR-TKI was significantly higher at the time of acquiring resistance than at baseline (p < 0.01).Conclusion: miR-21 is involved in acquired resistance of EGFR-TKI in NSCLC, which is mediated by down-regulating PTEN and PDCD4 and activating PI3K/Akt pathway. (C) 2013 Elsevier Ireland Ltd. All rights reserved.