Tumor-suppressive effects of microRNA-181d-5p on non-small-cell lung cancer through the CDKN3-mediated Akt signaling pathway in vivo and in vitro
Tumor-suppressive effects of microRNA-181d-5p on non-small-cell lung cancer through the CDKN3-mediated Akt signaling pathway in vivo and in vitro
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DOI：
10.1152/ajplung.00334.2018
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发表时间：
2019-05-01
影响因子：
4.9
通讯作者：
Xu, Shu-Feng
中科院分区：
文献类型：
--
作者：
Gao, Li-Ming;Zheng, Yue;Xu, Shu-Feng
一些microRNAs （miRs）通过抑制靶基因的表达参与肿瘤的发生和发展。然后通过微阵列分析筛选miR-181d-5p和细胞周期蛋白依赖性激酶抑制剂3 （CDKN3）在非小细胞肺癌（NSCLC）中的异常表达。在本研究中，我们进行了一系列体内和体外实验，旨在探讨它们在非小细胞肺癌中的作用及其潜在机制。CDKN3高表达，而miR-181d-5p在NSCLC中下调。定量RT-PCR、Western blot分析和双荧光素酶报告基因检测进一步证实了CDKN3可被miR-181d-5p负调控。此外，miR-181d-5p上调或CDKN3沉默可使Akt信号通路失活。采用5种NSCLC细胞系（H1299、A549、H1975、NCI-H157、GLC-82）中miR-181d-5p含量最低的A549和CDKN3含量最高的H1975进行体外实验，通过转染miR-181d-5p mimic/inhibitor或sirna靶向CDKN3改变miR-181d-5p和CDKN3的表达。然后，评估细胞增殖、凋亡、侵袭、迁移、血管生成以及上皮-间质转化（EMT），并评估致瘤性。此外，miR-181d-5p的升高或CDKN3的缺失导致NSCLC细胞的增殖、侵袭、迁移、血管生成、EMT和致瘤性的显著降低，并伴有细胞凋亡的增加。总之，本研究强调了miR-181d-5p通过抑制CDKN3使Akt信号通路失活对NSCLC的肿瘤抑制作用，从而为治疗NSCLC提供了一种有希望的治疗策略。
The involvement of several microRNAs (miRs) in the initiation and development of tumors through the suppression of the target gene expression has been highlighted. The aberrant expression of miR-181d-5p and cyclin-dependent kinase inhibitor 3 (CDKN3) in non-small-cell lung cancer (NSCLC) was then screened by microarray analysis. In the present study, we performed a series of in vivo and in vitro experiments for the purpose of investigating their roles in NSCLC and the underlying mechanism. There was a high expression of CDKN3, whereas miR-181d-5p was downregulated in NSCLC. Quantitative RT-PCR, Western blot analysis, and dual-luciferase reporter gene assay further identified that CDKN3 could be negatively regulated by miR-181d-5p. Moreover, the upregulation of miR-181d-5p or silencing of CDKN3 could inactivate the Akt signaling pathway. A549 with the lowest miR-181d-5p and H1975 with the highest CDKN3 among the five NSCLC cell lines (H1299, A549, H1975, NCI-H157, and GLC-82) were adopted for in vitro experiments, in which expression of miR-181d-5p and CDKN3 was altered by transfection of miR-181d-5p mimic/inhibitor or siRNA-targeting CDKN3. Afterwards, cell proliferation, apoptosis, invasion, migration, and angiogenesis, as well as epithelial-mesenchymal transition (EMT), were evaluated, and tumorigenicity was assessed. In addition, an elevation in miR-181d-5p or depletion in CDKN3 led to significant reductions in proliferation, invasion, migration, angiogenesis, EMT, and tumorigenicity of NSCLC cells, coupling with increased cell apoptosis. In conclusion, this study highlights the tumor-suppressive effects of miR-181d-5p on NSCLC via Akt signaling pathway inactivation by suppressing CDKN3, thus providing a promising therapeutic strategy for the treatment of NSCLC.