MicroRNA-181a regulates epithelial-mesenchymal transition by targeting PTEN in drug-resistant lung adenocarcinoma cells
MicroRNA-181a regulates epithelial-mesenchymal transition by targeting PTEN in drug-resistant lung adenocarcinoma cells
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DOI：
10.3892/ijo.2015.3144
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发表时间：
2015-10-01
影响因子：
5.2
通讯作者：
Liu, Xuegang
中科院分区：
文献类型：
--
作者：
Li, Haihui;Zhang, Pei;Liu, Xuegang
化疗耐药是肺腺癌中不可避免的现象，这与十号染色体上缺失的磷酸酶和张力蛋白同源物（PTEN）的表达降低有关。因此，确定抑制肺腺癌细胞化疗耐药性的新分子机制非常重要。紫杉醇和顺铂耐药的A549肺癌细胞衍生物是通过长期连续培养而开发的。使用伤口愈合测定、迁移测定、侵袭测定、形态学检查以及与上皮间质转化（EMT）相关的基因的蛋白质印迹分析/RT-PCR来评估细胞的转移特性。为了鉴定 A549 细胞中 EMT 的新调节因子，通过微阵列分析鉴定了耐药细胞中差异表达的 miRNA。通过用特定的模拟物和抑制剂转染，然后进行功能测定，确定了 miR-181a 的作用。进行荧光素酶测定以评估 miR-181a 靶向 PTEN 启动子的能力，并通过蛋白质印迹分析和 RT-PCR 评估 miR-181a 对 PTEN 表达的调节。与敏感细胞相比，紫杉醇和顺铂耐药的 A549 细胞获得了转移特性和 EMT 表型，并且 PTEN 表达减少。 miR-181a被鉴定为耐药A549细胞中差异表达的miRNA，并且miR-181a模拟物和抑制剂被证明影响EMT相关基因的迁移、侵袭、形态和表达。 PTEN 被确定为 miR-181a 的直接靶标。我们的研究结果表明，肺腺癌中的 miR-181a 表达可能通过靶向 PTEN 与 EMT 进展相关。 miR-181a的调节可能为克服肺腺癌中紫杉醇和顺铂的耐药性提供新的策略。
Chemoresistance is an inevitable occurrence in lung adenocarcinoma, which has been associated with decreased expression of the phosphatase and tensin homolog deleted on chromosome ten (PTEN). Therefore, it is important to identify novel molecular mechanisms to suppress chemoresistance in lung adenocarcinoma cells. Paclitaxel- and cisplatin-resistant A549 lung carcinoma cell derivatives were developed by long-term serial culture. The metastatic properties of the cells were assessed using wound-healing assays, migration assays, invasion assays, morphological examination, and western blot analysis/RT-PCR of genes associated with the epithelial-mesenchymal transition (EMT). To identify novel regulators of EMT in A549 cells, differentially expressed miRNAs in drug-resistant cells were identified by microarray analysis. The role of miR-181a was established by transfection with specific mimic and inhibitor followed by functional assays. Luciferase assays were performed to assess the ability of miR-181a to target the PTEN promoter, and regulation of PTEN expression by miR-181a was assessed by western blot analysis and RT-PCR. Paclitaxel- and cisplatin-resistant A549 cells acquired metastatic properties and EMT phenotype and had reduced PTEN expression as compared to sensitive cells. miR-181a was identified as a differentially expressed miRNA in drug-resistant A549 cells, and miR-181a mimic and inhibitor were shown to affect migration, invasion, morphology and expression of EMT-associated genes. PTEN was identified as a direct target of miR-181a. Our findings demonstrate that miR-181a expression in lung adenocarcinoma is associated with EMT progression, potentially through targeting of PTEN. Regulation of miR-181a may provide a novel strategy for overcoming resistance to paclitaxel and cisplatin in lung adenocarcinoma.