ABSTRACT
Deletion or mutation of the gene encoding the deubiquitinating enzyme CYLD is a common genomic aberration in multiple myeloma (MM). However, the functional consequence of CYLD loss and the mechanism underlying its putative role as a tumor suppressor gene in the pathogenesis of MM has not been established. Here, we show that CYLD expression is highly variable in myeloma cell lines and primary MMs and that low CYLD expression is associated with disease progression from monoclonal gammopathy of undetermined significance to MM, and with poor overall and progression free-survival of MM patients. Functional assays revealed that CYLD represses MM cell proliferation and survival. Furthermore, CYLD acts as a negative regulator of NF-κB and Wnt/ß-catenin signaling and loss of CYLD sensitizes MM cells to NF-κB-stimuli and Wnt ligands. Interestingly, in primary MMs, low CYLD expression strongly correlated with a proliferative and Wnt signaling-gene expression signature, but not with an NFκB target gene signature. Altogether, our findings identify CYLD as a negative regulator of NF-κB and Wnt/ß-catenin signaling in MM and indicate that loss of CYLD enhances MM aggressiveness through Wnt pathway activation. Thus, targeting the Wnt pathway could be a promising therapeutic strategy in MM with loss of CYLD activity.
Subject(s)
Multiple Myeloma/metabolism , Multiple Myeloma/pathology , Tumor Suppressor Proteins/deficiency , Wnt Signaling Pathway , Case-Control Studies , Deubiquitinating Enzyme CYLD , Humans , Multiple Myeloma/genetics , NF-kappa B/metabolism , Transfection , Tumor Suppressor Proteins/genetics , Tumor Suppressor Proteins/metabolismABSTRACT
Shwachman-Diamond syndrome (SDS) is a genetic disorder characterized by pancreatic hypoplasia, recurrent infection, and bone marrow (BM) dysfunction. SDS-patients have an increased frequency of myelodysplasia and leukemic transformation. Unspecific cytogenetic aberrations are a common finding in SDS. However, in a rising number of patients abnormalities of chromosome 7 have been reported, especially an i(7)(q10), which seems to be a non-random chromosome abnormality. Recently, the SDS gene has been mapped at locus 7q11 and subsequently cloned; recurrent mutations have been found. We report a case of SDS with an i(7)(q10) in the BM and two different mutations in the SBDS gene. At the age of 25 years, the patient suffers from mild aplastic anemia but does not show any clinical sign of myelodysplasia or leukemic transformation.
Subject(s)
Anemia, Aplastic/genetics , Chromosomes, Human, Pair 7 , Exocrine Pancreatic Insufficiency/genetics , Isochromosomes , Musculoskeletal Abnormalities/genetics , Proteins/genetics , Adult , Base Sequence , Disease Susceptibility , Female , Humans , In Situ Hybridization, Fluorescence , Karyotyping , Mutation , Myelodysplastic Syndromes/etiology , SyndromeABSTRACT
Classical cytogenetics has a low resolving power and allows analysis of dividing cells only. In fluorescence in situ hybridization (FISH), a DNA fragment is stained with a fluorescent marker, after which this fragment is brought into contact with a patient's DNA. The stained fragment can bind to a corresponding fragment, revealing its presence or absence. Using FISH, every desired DNA sequence (from a whole chromosome to one gene) can be stained. In this way it is also possible to diagnose microdeletion syndromes, such as the Williams syndrome, the DiGeorge syndrome and submicroscopic chromosome anomalies that play a part in mental handicaps. FISH also allows analysis of non-dividing cells. In this way it is possible for instance rapidly to examine uncultured amniotic fluid cells for the commoner trisomies or to find foetal erythrocytes in a pregnant woman's blood. It is also possible to demonstrate tumour-specific breaking points. By application of FISH to microarrays it is possible to study a large number of genes simultaneously for the presence of a particular number of DNA sequences linked to a clinical abnormality.
Subject(s)
Chromosome Aberrations/diagnosis , Chromosome Aberrations/genetics , DNA Mutational Analysis , In Situ Hybridization, Fluorescence/statistics & numerical data , Chromosome Deletion , Chromosome Disorders , Cytogenetic Analysis , DNA Probes , Humans , Microsatellite Repeats , Syndrome , Translocation, Genetic , Trisomy/diagnosis , Trisomy/geneticsABSTRACT
The locations of the genes encoding 18S, 5.8S and 28S rRNA and 5S rRNA were studied in two relatives of the domestic pig, the babirusa (Babyrousa babyrussa) and the white-lipped peccary (Tayassu pecari). In the babirusa, the 18S, 5.8S and 28S rDNA is located on chromosomes 6, 8 and 10. The genes on chromosomes 8 and 10 are actively transcribed, in contrast to those on chromosomes 6. In the white-lipped peccary, this rDNA was found to be located on chromosomes 4 and 8. The genes on both of these pairs of chromosomes are actively transcribed. The 5S rDNA was physically mapped to chromosome 16 in the babirusa, and to chromosome 11 in the white-lipped peccary. These data are compared to similar data obtained for the domestic pig, and confirm previously recognized chromosome homologies.
Subject(s)
Artiodactyla/genetics , DNA, Ribosomal/genetics , Animals , Artiodactyla/classification , Chromosome Banding , Chromosome Mapping , DNA Probes , In Situ Hybridization, Fluorescence , RNA, Ribosomal/genetics , RNA, Ribosomal, 18S/genetics , RNA, Ribosomal, 5.8S/genetics , RNA, Ribosomal, 5S/genetics , Species SpecificityABSTRACT
The gene clusters encoding 18S + 28S and 5S rRNA in the dog (Canis familiaris) have been localized by using GTG-banding and fluorescence in situ hybridization. The 18S + 28S rDNA maps to chromosome regions 7q2.5-->q2.7, 17q1.7, qter of a medium-sized, not yet numbered autosome, and Yq1.2-->q1.3. Our data show that there is one cluster of 5S rDNA in the dog, which maps to chromosome region 4q1.4.
Subject(s)
Chromosome Mapping , Dogs/genetics , RNA, Ribosomal, 18S/genetics , RNA, Ribosomal, 28S/genetics , RNA, Ribosomal, 5S/genetics , Animals , DNA, Ribosomal/analysis , Female , In Situ Hybridization, Fluorescence , MaleABSTRACT
A cDNA clone encoding the complete coding sequence for porcine alpha-1-antitrypsin (or alpha 1-protease inhibitor, PI) was isolated and its DNA sequence determined. The cDNA is assumed to encode alpha-1-antitrypsin on the basis of its sequence similarity to the corresponding cDNAs for human, baboon, rat, mouse, sheep and cow. The porcine cDNA clone was used in conjunction with BamHI, KpnI, MspI, SacI, TaqI and XbaI to develop restriction fragment length polymorphism-based genetic markers for linkage mapping in pigs. The cDNA has also been used to map the porcine PI locus to chromosome 7q2.4- > q2.6 by radioactive in situ hybridization. Thus, the PI locus has been added to the developing physical and genetic maps of the porcine genome.
Subject(s)
Chromosome Mapping , DNA, Complementary/genetics , Polymorphism, Restriction Fragment Length , Swine/genetics , alpha 1-Antitrypsin/genetics , Animals , Cloning, Molecular , Molecular Sequence Data , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Sequence Homology, Nucleic AcidABSTRACT
Homology among three pairs of domestic pig (Sus scrofa) and five pairs of babirusa (Babyrousa babyrussa) autosomes has been demonstrated with the use of porcine painting probes. With the results of this study, in addition to data obtained earlier through the application of banding techniques, correspondence between all individual chromosomes of these two distantly related pigs has been identified.
Subject(s)
Chromosomes , Swine/genetics , Animals , Coloring Agents , DNA Probes , Female , In Situ Hybridization, Fluorescence , Karyotyping , Male , Phylogeny , Species Specificity , Swine/classificationABSTRACT
Variation of the size of silver-stained nucleolar organizer regions (NORs) of chromosomes 10 and 8 was studied in pigs of six breeds (Sus scrofa L.). The silver deposits were quantified by image analysis and the results were normalized for each Ag-NOR chromosome. In general, normalized values for chromosomes 10 were higher than those for chromosomes 8, suggesting that the NOR activity of chromosomes 10 is dominant as compared to that of chromosomes 8. However, high values for chromosomes 8 were found in the Meishan breed and in some Piétrain pigs, indicating a high transcriptional activity of the rRNA genes on these chromosomes. In some pigs, the relative quantities of rDNA in chromosomes 10 and 8 were investigated by fluorescent in situ hybridization and the results were compared with those of the silver staining procedure. It is concluded that Ag-NOR sizes on chromosomes 10 are relatively well correlated to the number of rRNA genes, whereas the absence or the small size of Ag-NORs on chromosomes 8, often observed in pigs, is the result of low NOR activity rather than of absence of rDNA.
Subject(s)
Genetic Variation , Nucleolus Organizer Region/ultrastructure , RNA, Ribosomal/genetics , Swine/genetics , Animals , In Situ Hybridization, Fluorescence , Silver Staining , Species SpecificityABSTRACT
The genes coding for glucose regulated protein, 78kDal (GRP78), hormone-sensitive lipase (LIPE), plasminogen activator or urokinase (PLAU), and D-amino acid oxidase (DAO) were localized in the pig by radioactive in situ hybridization. GRP78 was mapped to 1q2.10-->q2.13 and LIPE was localized to chromosome 6cen-->q1.2. The genes for PLAU and DAO were both assigned to chromosome 14, in the region q2.4-->q2.6 and q2.1-->q2.3, respectively. The results are compared to mapping data in other mammalian species.
Subject(s)
Chromosome Mapping , Genetic Markers , Heat-Shock Proteins , Molecular Chaperones , Swine/genetics , Animals , Carrier Proteins/genetics , Cattle , D-Amino-Acid Oxidase/genetics , Endoplasmic Reticulum Chaperone BiP , Humans , In Situ Hybridization , Mice , Sterol Esterase/genetics , Urokinase-Type Plasminogen Activator/geneticsABSTRACT
Selective silver staining was used to investigate the cellular distribution of numbers of nucleolar organizer regions (NORs) in domestic pigs (Sus scrofa) of eight different breeds, the European wild boar (S. scrofa scrofa), Indonesian wild boar (S. scrofa vittatus), Javan warty pig (S. verrucosus), Sulawesi warty pig (S. celebensis), and pigmy hog (S. salvanius). In the domestic pig as well as in the wild (sub)species of Sus, actively transcribing ribosomal RNA genes were found to be present in the secondary constrictions of chromosome pairs 10 and 8. Chromosomes 10 were consistently Ag-positive. Chromosomes 8 less frequently showed Ag-NORs, resulting in different mean numbers of Ag-NORs per individual animal. Mean Ag-NOR numbers per breed or (sub)species were generally higher in the wild representatives of Sus than in the domestic breeds. The highest mean numbers of Ag-NORs were observed in the Meishan breed and in S. celebensis and S. salvanius. The Meishan breed appears to be conservative in Ag-NOR staining pattern, being more comparable to the Asian wild Suidae than to the European breeds.
Subject(s)
Nucleolus Organizer Region/chemistry , Swine/genetics , Animals , Animals, Domestic/genetics , Animals, Wild/genetics , Karyotyping , Nucleic Acid Hybridization , Silver StainingABSTRACT
Three cell lines derived from two metastases of a mammary carcinoma in a female dog were analyzed cytogenetically. All three cell lines showed a modal chromosome number of 76, with ranges of 74-77, 72-78, and 73-78. A biarmed chromosome in addition to the X chromosomes was observed in all cells of one cell line, and in a part of the cells of the other two cell lines. Results of banding analyses indicated that this chromosome was identical in the three cell lines, and can thus be considered a clonal marker. Additional biarmed chromosomes have not been reported previously from mammary tumor cells, although their presence is rather common in other canine neoplasms.
