Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis.
Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis.
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DOI：
10.1186/s13046-017-0571-8
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发表时间：
2017-08-03
期刊：
影响因子：
--
通讯作者：
Costanzo F
中科院分区：
文献类型：
--
作者：
Aversa I;Zolea F;Ieranò C;Bulotta S;Trotta AM;Faniello MC;De Marco C;Malanga D;Biamonte F;Viglietto G;Cuda G;Scala S;Costanzo F
铁蛋白在细胞内铁代谢中起核心作用;该分子是由24个重和轻类型亚基组成的纳米笼。重亚基（FHC）具有铁氧化酶活性，因此以无毒形式进行铁的关键转化。最近，已经表明FHC还参与另外的非铁相关的关键途径，包括p53调节、癌基因iRNA表达的调节和趋化因子信号传导。上皮细胞向间质细胞转化（EMT）是一种细胞机制，通过该机制，细胞获得成纤维细胞样表型沿着降低粘附力和增强运动性。在这项工作中，我们集中我们的注意力在人细胞系MCF-7和H460中FHC对EMT诱导的作用，以阐明潜在的分子机制。通过慢病毒驱动的shRNA策略进行FHC的靶向沉默。通过用Lipofectamine 2000转染全长FHC cDNA获得FHC基因产物的重建。MTT法和细胞计数法检测细胞活力和增殖能力;创伤愈合法和transwell法检测细胞迁移能力。通过流式细胞术进行CXCR 4表面表达的定量。实验数据表明，FHC沉默的MCF-7和H460细胞（MCF-7shFHC，H460 shFHC）获得间充质表型，伴随着其迁移和增殖能力的显着增强。这种转变与ROS产生的增加以及CXCR 4/CXCL 12信号通路的激活有关。我们目前的实验数据表明，ROS水平的胞浆增加是负责FHC沉默细胞的增殖增强，而更高的迁移率是由于CXCR 4/CXCL 12轴的失调。我们的研究结果表明，在MCF-7和H460细胞中，EMT的诱导、迁移和存活的增加依赖于FHC对两种途径的控制，即铁/ROS代谢和CXCR 4/CXCL 12轴。除了进一步证实FHC的多功能性质外，该数据还表明FHC量/功能的分析可能是预测肿瘤侵袭性的重要额外工具。
Ferritin plays a central role in the intracellular iron metabolism; the molecule is a nanocage of 24 subunits of the heavy and light types. The heavy subunit (FHC) is provided of a ferroxidase activity and thus performs the key transformation of iron in a non-toxic form. Recently, it has been shown that FHC is also involved in additional not iron-related critical pathways including, among the others, p53 regulation, modulation of oncomiRNAs expression and chemokine signalling. Epithelial to mesenchymal transition (EMT) is a cellular mechanism by which the cell acquires a fibroblast-like phenotype along with a decreased adhesion and augmented motility. In this work we have focused our attention on the role of the FHC on EMT induction in the human cell lines MCF-7 and H460 to elucidate the underlying molecular mechanisms. Targeted silencing of the FHC was performed by lentiviral-driven shRNA strategy. Reconstitution of the FHC gene product was obtained by full length FHC cDNA transfection with Lipofectamine 2000. MTT and cell count assays were used to evaluate cell viability and proliferation; cell migration capability was assayed by the wound-healing assay and transwell strategy. Quantification of the CXCR4 surface expression was performed by flow cytometry. Experimental data indicated that FHC-silenced MCF-7 and H460 cells (MCF-7shFHC, H460shFHC) acquire a mesenchymal phenotype, accompanied by a significant enhancement of their migratory and proliferative capacity. This shift is coupled to an increase in ROS production and by an activation of the CXCR4/CXCL12 signalling pathway. We present experimental data indicating that the cytosolic increase in ROS levels is responsible for the enhanced proliferation of FHC-silenced cells, while the higher migration rate is attributable to a dysregulation of the CXCR4/CXCL12 axis. Our findings indicate that induction of EMT, increased migration and survival depend, in MCF-7 and H460 cells, on the release of FHC control on two pathways, namely the iron/ROS metabolism and CXCR4/CXCL12 axis. Besides constituting a further confirmation of the multifunctional nature of FHC, this data also suggest that the analysis of FHC amount/function might be an important additional tool to predict tumor aggressiveness.
影响因子：
6.2
作者：
Abt, Anna Cook;Meucci, Olimpia
通讯作者：
Meucci, Olimpia