RESUMO
BACKGROUND: CYLD cutaneous syndrome (CCS; syn. Brooke-Spiegler syndrome) is a rare autosomal dominant hereditary disease characterized by multiple adnexal skin tumors including cylindromas, spiradenomas, and trichoepitheliomas. More than 100 germline mutations of the cylindromatosis (CYLD) gene have been reported in CCS and most of them are frameshift mutations or small alterations. METHODS: We identified a large, three-generation Chinese family with CCS, which consisted of 18 living family members, including six affected individuals. To explore the molecular biology of this family, we carried out targeted next-generation sequencing and Affymetrix CytoScan HD SNP array to analyze the mutation in the CYLD gene. RESULTS: A novel large deletion mutation, NC_000016.9:g.(50826498_50827517)_(50963389-50967346)del was found in the proband of this family. This deletion results in the loss of a nearly 140 kb fragment of the CYLD gene, spanning exons 17 ~ 20, which represent the coding regions of the ubiquitin-specific protease domain. Further quantitative polymerase chain reaction proved that all patients and two proband-related family members carried this large deletion. CONCLUSIONS: Our study expands the types of mutations in CCS and will undoubtedly provide valuable information for genetic counseling for families affected by the condition.
Assuntos
Enzima Desubiquitinante CYLD/genética , Síndromes Neoplásicas Hereditárias/genética , Neoplasias Cutâneas/genética , Adulto , Enzima Desubiquitinante CYLD/química , Feminino , Deleção de Genes , Heterozigoto , Humanos , Masculino , Síndromes Neoplásicas Hereditárias/patologia , Linhagem , Fenótipo , Domínios Proteicos , Neoplasias Cutâneas/patologiaRESUMO
Deubiquitinase cylindromatosis (CYLD) inhibits MAPK and NF-κB activation pathways by deubiquitinating upstream regulatory factors. Although CYLD has been identified and actively studied in mammals, nothing is known about its putative function in fish. In this study, we identified the gene encoding CYLD (OmCYLD) from rainbow trout, Oncorhynchus mykiss, and examined its role during pathogenic infections. The deduced amino acid sequence of OmCYLD contains conserved CAP-Gly and USP domains. In RTH-149 cells, the expression of OmCYLD was increased by stimulation with Edwardsiella tarda and Streptococcus iniae. Gain-of-function and loss-of-function experiments showed that OmCYLD down-regulates the activation of MAPK and NF-κB and the expression of pro-inflammatory cytokines in E. tarda-stimulated RTH-149 cells. These findings suggest that OmCYLD might function like those of mammals to negatively regulate bacteria-triggered signaling pathway in fish.
Assuntos
Enzima Desubiquitinante CYLD/genética , Enzima Desubiquitinante CYLD/imunologia , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Oncorhynchus mykiss/genética , Oncorhynchus mykiss/imunologia , Sequência de Aminoácidos , Animais , Enzima Desubiquitinante CYLD/química , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Filogenia , Alinhamento de Sequência/veterináriaRESUMO
OBJECTIVE: Gastric cancer is the second highest mortality tumor and the fourth most common cancer worldwide that has high aggressiveness. MicroRNA-181d (miR-181d) has been established to be a tumor suppressor, by suppressing cell proliferation, cell cycle, and promoting apoptosis in several cancers. The purpose of this study is to explore the great roles of miR-181d in gastric cancer. PATIENTS AND METHODS: The Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) and Western blot were applied to calculate the mRNA and protein levels of miR-181d and genes. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and transwell assays were utilized to measure the proliferative and invasive abilities. The Kaplan-Meier method was conducted to calculate the overall survival of gastric cancer patients. RESULTS: MiR-181d was detected to be downregulated in gastric cancer tissues and cell lines compared to the peritumoral normal tissues and normal cell line. Downregulation of miR-181d predicted poor prognosis of gastric cancer patients. Cylindromatosis gene (CYLD) was overexpressed in gastric cancer tissues, which was confirmed to be a target gene of miR-181d in gastric cancer cell line HGC-27. Moreover, miR-181d inhibited the proliferation through CYLD/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway and inhibited the invasion-mediated epithelial-mesenchymal transition (EMT) in HGC-27 cells. In addition, overexpression of miR-181d suppressed tumor growth and xenograft tumorigenesis of HGC-27 cells in vivo. CONCLUSIONS: MiR-181d functioned as a tumor suppressor by inhibiting the proliferation via PI3K/AKT pathway in vitro and in vivo and inhibiting invasion-mediated epithelial-mesenchymal transition (EMT) by targeting CYLD in gastric cancer. The newly identified miR-181d/CYLD axis provides novel insight into the pathogenesis of gastric cancer.
