Nicotine upregulates FGFR3 and RB1 expression and promotes non-small cell lung cancer cell proliferation and epithelial-to-mesenchymal transition via downregulation of miR-99b and miR-192
Nicotine upregulates FGFR3 and RB1 expression and promotes non-small cell lung cancer cell proliferation and epithelial-to-mesenchymal transition via downregulation of miR-99b and miR-192
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DOI：
10.1016/j.biopha.2018.02.113
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发表时间：
2018-05-01
影响因子：
7.5
通讯作者：
Han, Wei
中科院分区：
文献类型：
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作者：
Du, Xuemei;Qi, Fei;Han, Wei
背景：到目前为止，吸烟是非小细胞肺癌(NSCLC)的最大危险因素。尼古丁是烟草中的一种活性生物碱，不能启动人和啮齿动物的肿瘤发生，但可以促进烟草致癌物引发的包括非小细胞肺癌在内的各种肿瘤的生长和转移。最近，据报道，吸烟使大鼠肺内24个miRNAs下调了3倍以上，其中大多数下调的miRNAs与NSCLC的发生和发展有关。烟碱作为烟草的主要成分可能与一些miRNA的表达变化有关。方法：采用QRT-PCR方法检测miRNA和mRNAs的表达，Western印迹法检测蛋白表达。结果：检测尼古丁对NSCLC细胞中24个miRNAs表达的影响，结果显示尼古丁处理NSCLC后，miR-99b和miR-192的表达均下降。细胞增殖和上皮细胞间充质转化(EMT)检测显示，尼古丁促进NSCLC细胞增殖和EMT，而miR-99b或miR-192表达的恢复可缓解尼古丁对NSCLC细胞增殖和EMT的影响。随后，成纤维细胞生长因子受体3(FGFR3)和视网膜母细胞瘤1(RB1)分别被证实为miR-99b和miR-192的靶点，并被尼古丁上调。结论：本研究首次阐明尼古丁-miR-99b/miR-192-FGFR3/RB1调控网络是尼古丁通过下调miR-99b和miR-192，上调其靶标FGFR3和RB1，促进NSCLC细胞增殖和EMT。这些发现为理解非小细胞肺癌与尼古丁效应相关的潜在分子机制提供了新的见解。
Background: Tobacco smoke is by far the greatest risk factor for non-small-cell lung cancer (NSCLC). Nicotine, an active alkaloid in tobacco, is unable to initiate tumorigenesis in humans and rodents, but can promote the growth and metastasis of various tumors, including NSCLC, initiated by tobacco carcinogens. Recently, cigarette smoke is reported to downregulate 24 miRNAs more than 3-fold in the lungs of rats, and most of these downregulated miRNAs are associated with NSCLC initiation and development. Nicotine as the major tobacco component might be associated with the expression changes of some miRNAs.Methods: qRT-PCR was performed to determine the miRNA and mRNA expression, and western blot was conducted to measure protein expression. MTT assay was used to detect cell proliferation.Results: The effects of nicotine on the expression of 24 miRNAs in NSCLC cell lines were determined, and the results showed that nicotine treatment decreased miR-99b and miR-192 expression. Cell proliferation and epithelial-to-mesenchymal transition (EMT) detection showed that nicotine promoted NSCLC cell proliferation and EMT, and restoration of miR-99b or miR-192 expression relieved the effects of nicotine on NSCLC cell proliferation and EMT. Subsequently, fibroblast growth factor receptor 3 (FGFR3) and retinoblastoma 1 (RB1) were confirmed to be the targets of miR-99b and miR-192, respectively, and were upregulated by nicotine in NSCLC cells. In addition, FGFR3 or RB1 knockdown inhibited NSCLC cell proliferation and EMT.Conclusion: This study, for the first time, elucidates nicotine-miR-99b/miR-192-FGFR3/RB1 regulatory network that nicotine promotes NSCLC cell proliferation and EMT by downregulating miR-99b and miR-192, and upregulating their targets FGFR3 and RB1. These findings offer novel insights into the understanding of underlying molecular mechanisms of NSCLC related with the nicotine effects.