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 H subunit (FHC), is involved in different biological events such as cell differentiation and pathologic states such as neurodegeneration and cancer. This study is aimed at investigating the wholecell proteome of the FHC-silenced MM07m cells, a cell line established from a supraclavicular lymph node metastasis, in comparison with that of the wild-type counterpart, in the attempt to identify and classify the highest number of proteins directly or indirectly controlled by the FHC. More than 500 peptides were identified and classified 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 processes related to tumor progression and metastasis. Among them: galectin-1, a family of proteins that is essential in tumor angiogenesis, whose overexpression is often related to invasiveness; cathepsin B, that is involved in cancer metastasis by facilitating cell invasiveness and migration; integrin-a5, which is known to play a critical role in cell surface-mediated signaling and adhesion functions. In vitro assays have confirmed that the FHC-silenced MM07m cells are characterized by a decreased growth activity, by a reduced invasiveness and a reduced cell adhesion capability. Moreover nude mice (CD1 nu/nu), subcutaneously-injected with FHC-silenced MM07m cells, have shown a remarkable 4-fold reduction of their tumor growth capacity compared to those who received the FHC-unsilenced MM07m counterpart. In conclusion, the coupling of gene silencing technology with the proteomic analysis appears as an original and very promising approach to identify the molecular patterns in which the silenced gene might be involved.
The proteome of the FHC silenced cell : in vitro and in vivo analysis / R. Misaggi, M. Di Sanzo, M. Gaspari, F. Romeo, L. Falbo, G. Cuda, B. Quaresima, M.C. Faniello, F. Costanzo. ((Intervento presentato al 36. convegno FEBS Congress : Biochemistry for tomorrow's medicine tenutosi a Torino nel 2011.
The proteome of the FHC silenced cell : in vitro and in vivo analysis
F. Romeo;L. Falbo;
2011
Abstract
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 H subunit (FHC), is involved in different biological events such as cell differentiation and pathologic states such as neurodegeneration and cancer. This study is aimed at investigating the wholecell proteome of the FHC-silenced MM07m cells, a cell line established from a supraclavicular lymph node metastasis, in comparison with that of the wild-type counterpart, in the attempt to identify and classify the highest number of proteins directly or indirectly controlled by the FHC. More than 500 peptides were identified and classified 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 processes related to tumor progression and metastasis. Among them: galectin-1, a family of proteins that is essential in tumor angiogenesis, whose overexpression is often related to invasiveness; cathepsin B, that is involved in cancer metastasis by facilitating cell invasiveness and migration; integrin-a5, which is known to play a critical role in cell surface-mediated signaling and adhesion functions. In vitro assays have confirmed that the FHC-silenced MM07m cells are characterized by a decreased growth activity, by a reduced invasiveness and a reduced cell adhesion capability. Moreover nude mice (CD1 nu/nu), subcutaneously-injected with FHC-silenced MM07m cells, have shown a remarkable 4-fold reduction of their tumor growth capacity compared to those who received the FHC-unsilenced MM07m counterpart. In conclusion, the coupling of gene silencing technology with the proteomic analysis appears as an original and very promising approach to identify the molecular patterns in which the silenced gene might be involved.
Pubblicazioni consigliate
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
