Identification of H ferritin-dependent and independent genes in K562 differentiating cells by targeted gene silencing and expression profiling.
Ferritin is best known as the key molecule in intracellular iron storage, and is involved in several metabolic processes such as cell proliferation, differentiation and neoplastic transformation. We have recently demonstrated that the shRNA silencing of the ferritin heavy subunit (FHC) in a melanoma cell line is accompanied by a consistent modification of gene expression pattern leading to a reduced potential in terms of proliferation, invasiveness, and adhesion ability of the silenced cells. In this study we sought to define the repertoire of genes whose expression might be affected by FHC during the hemin-induced differentiation of the erythromyeloid cell line K562. To this aim, gene expression profiling was performed in four different sets of cells: i) wild type K562; ii) sh-RNA FHC-silenced K562; iii) hemin-treated wild-type K562; and iv) hemin-treated FHC-silenced K562. Statistical analysis of the gene expression data, performed by two-factor ANOVA, identified three distinct classes of transcripts: a) Class 1, including 657 mRNAs whose expression is modified exclusively during hemin-induced differentiation of K562 cells, independently from the FHC relative amounts; b) Class 2, containing a set of 70 mRNAs which are consistently modified by hemin and FHC-silencing; and c) Class 3, including 128 transcripts modified by FHC-silencing but not by hemin. Our data indicate that FHC may function as a modulator of gene expression during erythroid differentiation and add new findings to the knowledge of the complex gene network modulated during erythroid differentiation.
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Roberta
R
Misaggi
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Maddalena
M
Di Sanzo
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Carlo
C
Cosentino
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Heather M
HM
Bond
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Domenica
D
Scumaci
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Francesco
F
Romeo
Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, via Allende, 84081 Baronissi, Salerno, Italy.
Claudia
C
Stellato
Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, via Allende, 84081 Baronissi, Salerno, Italy.
Giorgio
G
Giurato
Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, via Allende, 84081 Baronissi, Salerno, Italy.
Alessandro
A
Weisz
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Barbara
B
Quaresima
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Tullio
T
Barni
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Francesco
F
Amato
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Giuseppe
G
Viglietto
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Giovanni
G
Morrone
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Giovanni
G
Cuda
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy. Electronic address: faniello@unicz.it.
Maria Concetta
MC
Faniello
Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Viale Europa, 88100 Catanzaro, Italy.
Francesco
F
Costanzo
D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D016000:Cluster Analysis; D019295:Computational Biology; D005293:Ferritins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D053263:Gene Regulatory Networks; D020868:Gene Silencing; D006427:Hemin / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D020014:K562 Cells; D021122:Protein Subunits / Q000235:genetics; D034622:RNA Interference; D015398:Signal Transduction
D021122:Protein Subunits; D006427:Hemin; D005293:Ferritins
Differentiation; FHC; FLC; Ferritin Heavy Chain; Ferritin Light Chain; Gene expression; Microarray; qPCR; quantitative Real Time Polymerase Chain Reaction; shRNA; short hairpin RNA