MiR-16 regulates the pro-tumorigenic potential of lung fibroblasts through the inhibition of HGF production in an FGFR-1- and MEK1-dependent manner.
MiR-16 regulates the pro-tumorigenic potential of lung fibroblasts through the inhibition of HGF production in an FGFR-1- and MEK1-dependent manner.
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DOI：
10.1186/s13045-018-0594-4
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发表时间：
2018-03-20
影响因子：
28.5
通讯作者：
Lecis D
中科院分区：
文献类型：
--
作者：
Andriani F;Majorini MT;Mano M;Landoni E;Miceli R;Facchinetti F;Mensah M;Fontanella E;Dugo M;Giacca M;Pastorino U;Sozzi G;Delia D;Roz L;Lecis D
成纤维细胞通过细胞外基质的合成和重塑以及多种可溶性因子的产生，是肿瘤-基质相互作用的重要介质。尽管如此，对于肺癌中成纤维细胞促肿瘤活性的具体决定因素仍然知之甚少。在这里，我们的目的是了解 miRNA 在将成纤维细胞重编程为肿瘤支持表型方面的作用，这些 miRNA 在基质细胞中经常发生改变。我们采用基于共培养的高通量筛选来鉴定调节肺成纤维细胞促肿瘤发生潜力的特定 miRNA。多重测定和 ELISA 有助于研究 miRNA 对肺癌患者原代肺成纤维细胞和永生化肺成纤维细胞分泌组的影响，以及评估重度吸烟者血浆 HGF 水平。通过双荧光素酶报告基因测定和蛋白质印迹研究了 miRNA 的直接 mRNA 靶向。最后，通过体外迁移实验和小鼠异种移植物测试了成纤维细胞及其条件培养基的促肿瘤活性。我们确定 miR-16 是成纤维细胞分泌组的主要调节因子，并表明其上调会减少成纤维细胞的 HGF 分泌，从而损害其促进癌细胞迁移的能力。这种效应是由于 miR-16 的多效性活性所致，它通过直接抑制 FGFR-1 信号传导和靶向 HGF mRNA 来阻止 HGF 表达。从机制上讲，miR-16 靶向 FGFR-1 下游介质 MEK1，从而减少 ERK1/2 激活。一致地，FGFR-1 的化学或遗传抑制模拟 miR-16 活性并阻止 HGF 产生。值得注意的是，我们报告说，与未接触烟雾的肺部相比，来自重度吸烟者肺部的原代成纤维细胞系表达的 miR-16 水平降低，并且重度吸烟者血浆中的 HGF 浓度与烟草暴露水平相关。最后，体内实验证实，成纤维细胞中miR-16表达的恢复降低了其促进肿瘤生长的能力，并且HGF在成纤维细胞的促肿瘤活性中发挥着核心作用。总体而言，这些结果揭示了 miR-16 在调节肺成纤维细胞 HGF 产生中的核心作用，从而影响其促肿瘤潜力。吸烟暴露与 miR-16 水平之间的相关性可以为肺癌发展早期阶段肿瘤相关环境的形成提供新的线索。本文的在线版本 (10.1186/s13045-018-0594-4) 包含补充材料，可供授权用户使用。
Fibroblasts are crucial mediators of tumor-stroma cross-talk through synthesis and remodeling of the extracellular matrix and production of multiple soluble factors. Nonetheless, little is still known about specific determinants of fibroblast pro-tumorigenic activity in lung cancer. Here, we aimed at understanding the role of miRNAs, which are often altered in stromal cells, in reprogramming fibroblasts towards a tumor-supporting phenotype. We employed a co-culture-based high-throughput screening to identify specific miRNAs modulating the pro-tumorigenic potential of lung fibroblasts. Multiplex assays and ELISA were instrumental to study the effect of miRNAs on the secretome of both primary and immortalized lung fibroblasts from lung cancer patients and to evaluate plasmatic levels of HGF in heavy smokers. Direct mRNA targeting by miRNAs was investigated through dual-luciferase reporter assay and western blot. Finally, the pro-tumorigenic activity of fibroblasts and their conditioned media was tested by employing in vitro migration experiments and mouse xenografts. We identified miR-16 as a master regulator of fibroblast secretome and showed that its upregulation reduces HGF secretion by fibroblasts, impairing their capacity to promote cancer cell migration. This effect is due to a pleiotropic activity of miR-16 which prevents HGF expression through direct inhibition of FGFR-1 signaling and targeting of HGF mRNA. Mechanistically, miR-16 targets FGFR-1 downstream mediator MEK1, thus reducing ERK1/2 activation. Consistently, chemical or genetic inhibition of FGFR-1 mimics miR-16 activity and prevents HGF production. Of note, we report that primary fibroblast cell lines derived from lungs of heavy smokers express reduced miR-16 levels compared to those from lungs not exposed to smoke and that HGF concentration in heavy smokers’ plasma correlates with levels of tobacco exposure. Finally, in vivo experiments confirmed that restoration of miR-16 expression in fibroblasts reduced their ability to promote tumor growth and that HGF plays a central role in the pro-tumorigenic activity of fibroblasts. Overall, these results uncover a central role for miR-16 in regulating HGF production by lung fibroblasts, thus affecting their pro-tumorigenic potential. Correlation between smoking exposure and miR-16 levels could provide novel clues regarding the formation of a tumor-proficient milieu during the early phases of lung cancer development. The online version of this article (10.1186/s13045-018-0594-4) contains supplementary material, which is available to authorized users.
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影响因子：
37.3
作者：
Frediani JN;Fabbri M
通讯作者：
Fabbri M
影响因子：
82.9
作者：
Bonci, Desiree;Coppola, Valeria;De Maria, Ruggero
通讯作者：
De Maria, Ruggero
影响因子：
6.2
作者：
Kim SM;Kim H;Yun MR;Kang HN;Pyo KH;Park HJ;Lee JM;Choi HM;Ellinghaus P;Ocker M;Paik S;Kim HR;Cho BC
通讯作者：
Cho BC
DOI：
10.1084/jem.20140692
发表时间：
2014-07-28
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Öhlund D;Elyada E;Tuveson D
通讯作者：
Tuveson D
影响因子：
6.6
作者：
Andriani F;Bertolini G;Facchinetti F;Baldoli E;Moro M;Casalini P;Caserini R;Milione M;Leone G;Pelosi G;Pastorino U;Sozzi G;Roz L
通讯作者：
Roz L