title: C-type natriuretic peptide in combination with sildenafil attenuates proliferation of rhabdomyosarcoma cells.
abstract: Rhabdomyosarcoma (RMS) is a malignant mesenchymal tumor and the most common soft tissue sarcoma in children. Because of several complications associated with intensive multimodal therapies, including growth disturbance and secondary cancer, novel therapies with less toxicity are urgently needed. C-type natriuretic peptide (CNP), an endogenous peptide secreted by endothelial cells, exerts antiproliferative effects in multiple types of mesenchymal cells. Therefore, we investigated whether CNP attenuates proliferation of RMS cells. We examined RMS patient samples and RMS cell lines. All RMS clinical samples expressed higher levels of guanylyl cyclase B (GC-B), the specific receptor for CNP, than RMS cell lines. GC-B expression in RMS cells decreased with the number of passages in vitro. Therefore, GC-B stable expression lines were established to mimic clinical samples. CNP increased cyclic guanosine monophosphate (cGMP) levels in RMS cells in a dose-dependent manner, demonstrating the biological activity of CNP. However, because cGMP is quickly degraded by phosphodiesterases (PDEs), the selective PDE5 inhibitor sildenafil was added to inhibit its degradation. In vitro, CNP, and sildenafil synergistically inhibited proliferation of RMS cells stably expressing GC-B and decreased Raf-1, Mitogen-activated protein kinase kinase (MEK), and extracellular signal-regulated kinase (ERK) phosphorylation. These results suggested that CNP in combination with sildenafil exerts antiproliferative effects on RMS cells by inhibiting the Raf/MEK/ERK pathway. This regimen exerted synergistic effects on tumor growth inhibition without severe adverse effects in vivo such as body weight loss. Thus, CNP in combination with sildenafil represents a promising new therapeutic approach against RMS.
authors.affiliation: Department of Biochemistry, National Cerebral and Cardiovascular Center Research Institute, Suita-City, Osaka, Japan.|Department of Pediatric Surgery, Osaka University Graduate School of Medicine, Suita-City, Osaka, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Zenitani
authors.affiliation: Department of Biochemistry, National Cerebral and Cardiovascular Center Research Institute, Suita-City, Osaka, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Nojiri
authors.affiliation: Department of Pediatric Surgery, Osaka University Graduate School of Medicine, Suita-City, Osaka, Japan.
authors.forename: Shuichiro
authors.initials: S
authors.lastname: Uehara
authors.affiliation: Department of Biochemistry, National Cerebral and Cardiovascular Center Research Institute, Suita-City, Osaka, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Miura
authors.affiliation: Departments of Regenerative Medicine and Tissue Engineering, National Cerebral and Cardiovascular Center Research Institute, Suita-City, Osaka, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hosoda
authors.affiliation: Department of Biochemistry, National Cerebral and Cardiovascular Center Research Institute, Suita-City, Osaka, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Kimura
authors.affiliation: Department of Pediatric Surgery, Osaka University Graduate School of Medicine, Suita-City, Osaka, Japan.
authors.forename: Kengo
authors.initials: K
authors.lastname: Nakahata
authors.affiliation: Department of Biochemistry, National Cerebral and Cardiovascular Center Research Institute, Suita-City, Osaka, Japan.
authors.forename: Mikiya
authors.initials: M
authors.lastname: Miyazato
authors.affiliation: Department of Pediatric Surgery, Osaka University Graduate School of Medicine, Suita-City, Osaka, Japan.
authors.forename: Hiroomi
authors.initials: H
authors.lastname: Okuyama
authors.affiliation: Department of Biochemistry, National Cerebral and Cardiovascular Center Research Institute, Suita-City, Osaka, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Kangawa
mesh_terms: D000293:Adolescent; D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D002648:Child; D002675:Child, Preschool; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005260:Female; D006801:Humans; D007223:Infant; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008297:Male; D008807:Mice, Inbred BALB C; D020098:Natriuretic Peptide, C-Type / Q000008:administration & dosage; D010766:Phosphorylation / Q000187:drug effects; D017461:Receptors, Atrial Natriuretic Factor / Q000378:metabolism; D012208:Rhabdomyosarcoma / Q000201:enzymology / Q000473:pathology*; D000068677:Sildenafil Citrate / Q000008:administration & dosage; D014407:Tumor Cells, Cultured; D015854:Up-Regulation; D023041:Xenograft Model Antitumor Assays / Q000379:methods; D000099242:Atrial Natriuretic Factor Receptor B
chemical_list: D020098:Natriuretic Peptide, C-Type; D017461:Receptors, Atrial Natriuretic Factor; D000068677:Sildenafil Citrate; D000099242:Atrial Natriuretic Factor Receptor B
keywords: Antiproliferation; CNP; ERK; PDE5 inhibitor; cGMP
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