title: Expression and Function of Phosphodiesterase Type 5 in Human Breast Cancer Cell Lines and Tissues: Implications for Targeted Therapy.
abstract: By catalyzing cGMP hydrolysis, phosphodiesterase (PDE) 5 is a critical regulator of its concentration and effects in different (patho)physiologic processes, including cancers. As PDE5 is a known druggable target, we investigated the clinical significance of its expression in breast cancer and the underlying mechanisms by which it may contribute to tumor progression.
  
  
  
  PDE5 expression was evaluated in seven breast cancer cell lines by RT-PCR and immunoblotting. To examine the impact of PDE5 on cancer phenotype, MCF-7 cells expressing lower levels of the enzyme were engineered to stably overexpress PDE5. Proliferation was evaluated by MTT assays, motility and invasion by wound-healing/transmigration/invasion assays, transcriptome-profiling by RNA-sequencing, and Rho GTPase signaling activation by GST-pulldown assays and immunoblotting. Clinical relevance was investigated by IHC on tissues and retrospective studies from METABRIC cohort.
  
  
  
  PDE5 is differentially expressed in each molecular subtype of both breast cancer cell lines and tissues, with higher levels representing a startling feature of HER2-positive and triple-negative breast cancers. A positive correlation was established between elevated PDE5 levels and cancers of high histologic grade. Higher PDE5 expression correlated with shorter patient survival in retrospective analyses. On molecular level, stable PDE5 overexpression in Luminal-A-like MCF-7 cells resulted in enhanced motility and invasion through Rho GTPase signaling activation. Treatment of PDE5-stable clones with selective ROCK or PDE5 inhibitors completely restored the less motile and weak invasive behavior of control vector cells.
  
  
  
  PDE5 expression enhances breast cancer cell invasive potential, highlighting this enzyme as a novel prognostic candidate and an attractive target for future therapy in breast cancers. Clin Cancer Res; 22(9); 2271-82. ©2015 AACR.
authors.affiliation: Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Rende (CS), Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Catalano
authors.affiliation: Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Rende (CS), Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Campana
authors.affiliation: Centro Sanitario, University of Calabria, Rende (CS), Italy.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Giordano
authors.affiliation: MTA TTK Lendület Cancer Biomarker Research Group, 2nd Department of Pediatrics, Semmelweis University, Budapest, Hungary.
authors.forename: Balázs
authors.initials: B
authors.lastname: Győrffy
authors.affiliation: Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Tarallo
authors.affiliation: Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Rinaldi
authors.affiliation: Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy.
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Bruno
authors.affiliation: Division of Anatomic Pathology, Annunziata Hospital, Cosenza (CS), Italy.
authors.forename: Aurora
authors.initials: A
authors.lastname: Ferraro
authors.affiliation: Division of Anatomic Pathology, Annunziata Hospital, Cosenza (CS), Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Romeo
authors.affiliation: Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Rende (CS), Italy.
authors.forename: Marilena
authors.initials: M
authors.lastname: Lanzino
authors.affiliation: Department of Anatomical, Histological, Forensic, and Orthopedic Sciences, Sapienza University, Rome, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Naro
authors.affiliation: Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Rende (CS), Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Bonofiglio
authors.affiliation: Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Rende (CS), Italy. Centro Sanitario, University of Calabria, Rende (CS), Italy. inesbarone@virgilio.it sebastiano.ando@unical.it.
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Andò
authors.affiliation: Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Rende (CS), Italy. inesbarone@virgilio.it sebastiano.ando@unical.it.
authors.forename: Ines
authors.initials: I
authors.lastname: Barone
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000502:physiology; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5 / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D006801:Humans; D061986:MCF-7 Cells; D008875:Middle Aged; D015398:Signal Transduction / Q000502:physiology; D059467:Transcriptome / Q000502:physiology; D020741:rho GTP-Binding Proteins / Q000378:metabolism
chemical_list: D054706:Cyclic Nucleotide Phosphodiesterases, Type 5; D020741:rho GTP-Binding Proteins