title: Effects of inhibition of hedgehog signaling on cell growth and migration of uveal melanoma cells.
abstract: The Hedgehog (Hh) signaling pathway has been demonstrated to play a critical role in controlling embryonic development, tissue patterning, wound healing and a variety of cell functions. Aberrant activation of Hh signaling is implicated in the pathogenesis of many human cancers, and in angiogenesis. However, the role of this pathway in uveal melanoma (UM) carcinogenesis remains unknown. In this study, we investigated the effects of Hh inhibition using the specific Smoothened (Smo) antagonist cyclopamine to block Hh signaling in cultured human UM cell lines expressing Hh signaling components. Cyclopamine treatment effectively increased apoptosis and inhibited cell proliferation, migration, and epithelial-to-mesenchymal transition (EMT) by downregulating the Hh final arbiter glioblastoma 1 (Gli1), which regulates the transcription of target genes in the nucleus. Changes in gene and protein expression levels were detected by real-time PCR and by western blotting and immunocytochemistry, respectively. Cell cycle and apoptosis regulation induced by cyclopamine were demonstrated by flow cytometry. In addition, the migration capability of UM cells was reduced, as demonstrated by transwell migration and scratch assays. The effects of Hh inhibition on the levels of angiogenesis factors secreted by UM cells were examined by tube-formation assay.
  
  
  CONCLUSIONS
  Blocking the Hh pathway by cyclopamine decreased cell viability, migration, EMT, and angiogenesis, increased apoptosis, and induced G 1 phase cell cycle arrest in UM cells. Collectively, these results provide the first evidence of the significance of Gli1 activation downstream of Smo as a therapeutic target and the potential value of cyclopamine for the treatment of human UM.
authors.affiliation: Department of Ophthalmology; Ninth People's Hospital; Shanghai Jiaotong University School of Medicine; Shanghai, PR China.
authors.forename: Fei
authors.initials: F
authors.lastname: Duan
authors.affiliation: Department of Ophthalmology; Ninth People's Hospital; Shanghai Jiaotong University School of Medicine; Shanghai, PR China.
authors.forename: Ming
authors.initials: M
authors.lastname: Lin
authors.affiliation: Department of Ophthalmology; Ninth People's Hospital; Shanghai Jiaotong University School of Medicine; Shanghai, PR China.
authors.forename: Chuanyin
authors.initials: C
authors.lastname: Li
authors.affiliation: Department of Ophthalmology; Ninth People's Hospital; Shanghai Jiaotong University School of Medicine; Shanghai, PR China.
authors.forename: Xia
authors.initials: X
authors.lastname: Ding
authors.affiliation: Department of Ophthalmology; Ninth People's Hospital; Shanghai Jiaotong University School of Medicine; Shanghai, PR China.
authors.forename: Guanxiang
authors.initials: G
authors.lastname: Qian
authors.affiliation: Department of Ophthalmology; Ninth People's Hospital; Shanghai Jiaotong University School of Medicine; Shanghai, PR China.
authors.forename: He
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Ophthalmology; Ninth People's Hospital; Shanghai Jiaotong University School of Medicine; Shanghai, PR China.
authors.forename: Shengfang
authors.initials: S
authors.lastname: Ge
authors.affiliation: Department of Ophthalmology; Ninth People's Hospital; Shanghai Jiaotong University School of Medicine; Shanghai, PR China.
authors.forename: Xianqun
authors.initials: X
authors.lastname: Fan
authors.affiliation: Department of Ophthalmology; Ninth People's Hospital; Shanghai Jiaotong University School of Medicine; Shanghai, PR China.
authors.forename: Jin
authors.initials: J
authors.lastname: Li
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects; D053823:Hedgehog Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D008545:Melanoma / Q000473:pathology*; D034741:RNA, Small Interfering / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000037:antagonists & inhibitors / Q000378:metabolism; D015398:Signal Transduction; D000072086:Smoothened Receptor; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014604:Uveal Neoplasms / Q000473:pathology*; D014704:Veratrum Alkaloids / Q000494:pharmacology*; D000071676:Zinc Finger Protein GLI1; D000098943:Uveal Melanoma
chemical_list: C400551:GLI1 protein, human; D053823:Hedgehog Proteins; D034741:RNA, Small Interfering; D043562:Receptors, G-Protein-Coupled; C491023:SMO protein, human; D000072086:Smoothened Receptor; D014157:Transcription Factors; D014704:Veratrum Alkaloids; D000071676:Zinc Finger Protein GLI1; C000541:cyclopamine
keywords: Hedgehog signaling pathway; cell growth; cell migration; cyclopamine; uveal melanoma