H Ferritin Gene Silencing in a Human Metastatic Melanoma Cell Line: A Proteomic Analysis
H Ferritin Gene Silencing in a Human Metastatic Melanoma Cell Line: A Proteomic Analysis
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DOI：
10.1021/pr200705z
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发表时间：
2011-12-01
影响因子：
4.4
通讯作者：
Faniello, Maria Concetta
中科院分区：
文献类型：
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作者：
Di Sanzo, Maddalena;Gaspari, Marco;Faniello, Maria Concetta
铁蛋白是细胞内主要的铁储存蛋白，由24个亚基组成，分为H和L两种类型。除了调节细胞内铁稳态之外，已经发现铁蛋白，特别是H亚基（FHC），参与不同的生物学事件，例如细胞分化和病理状态（即，神经变性和癌症）。本研究旨在研究表达FHC和sh-RNA沉默的人转移性黑色素瘤细胞（MM 07（m））的全细胞蛋白质组，以识别和分类直接或间接受FHC控制的最大数量的蛋白质。我们确定了约200个差异表达的蛋白质，并根据其功能将其归类为参与代谢过程、细胞粘附、迁移和增殖过程的蛋白质。其中一些引起了我们的注意，因为它们参与了与肿瘤进展和转移相关的代谢途径。体外试验证实，FHC沉默的MM 07（m）细胞的特征在于生长活性降低、侵袭性降低和细胞粘附能力降低。此外，皮下注射FHC沉默的MM 07（m）细胞的裸鼠（CD 1 nu/nu）显示，与接受FHC未沉默的MM 07（m）对应物的裸鼠相比，其肿瘤生长能力显著降低4倍。总之，这些数据表明，基因沉默技术，再加上蛋白质组学分析，是一个强大的工具，更好地了解H铁蛋白信号转导通路，并提供支持的假设，该基因的特异性靶向可能是一个有吸引力的和潜在的有效的战略管理转移性黑色素瘤。
Ferritin, the major intracellular iron-storage protein, is made of 24 subunits of two types, H and L. Besides regulating intracellular iron homeostasis, it has been found that ferritin, in particular the H subunit (FHC), is involved in different biological events such as cell differentiation and pathologic states (i.e., neurodegeneration and cancer). This study is aimed at investigating the whole-cell proteome of FHC-expressing and sh-RNA-silenced human metastatic melanoma cells (MM07(m)) in the attempt to identify and classify the highest number of proteins directly or indirectly controlled by the FHC. We identified about 200 differentially expressed proteins and classified them in clusters on the basis of their functions, as proteins involved in metabolic processes, cell adhesion, migration, and proliferation processes. Some of them have captured our attention because of their involvement in metabolic pathways related to tumor progression and metastasis. In vitro assays confirmed that the FHC-silenced MM07(m) cells are characterized by a decreased growth activity, a reduced invasiveness, and a reduced cell adhesion capability. Moreover, nude mice (CD1 nu/nu), subcutaneously injected with FHC-silenced MM07(m) cells, showed a remarkable 4-fold reduction of their tumor growth capacity compared to those who received the FHC-unsilenced MM07(m) counterpart. In conclusion, these data indicate that gene silencing technology, coupled to proteomic analysis, is a powerful tool for a better understanding of H ferritin signaling pathways and lend support to the hypothesis that specific targeting of this gene might be an attractive and potentially effective strategy for the management of metastatic melanoma.