RESUMO
The Drosophila spinster (spin) mutant was isolated as a mutant that showed abnormal morphology and function in the nervous system. The spin defect induces neural degeneration similar to human lysosomal storage diseases. Various studies have shown that Spin proteins are localized in lysosomes and participate in the late stages of the autophagic process. Vertebrates have three spinster orthologs, Spns1, Spns2, and Spns3. A defect in Spns1 caused a short lifespan with aberrant lysosomal function in zebrafish. Spns2 was originally isolated as the gene responsible for abnormal heart development and was identified as a sphingosine 1-phosphate transporter in zebrafish. An endothelial cell-specific defect in Spns2 resulted in impaired egress of lymphocytes and the prevention of tumor metastasis in mice. Herein, I reviewed the history of spin/Spns research and discussed the conserved and newly diverged spin/Spns function and possible implications for human diseases.
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Glicoproteínas de Membrana , Proteínas Secretadas Inibidoras de Proteinases , Animais , Autofagia/fisiologia , Humanos , Lisofosfolipídeos/metabolismo , Lisossomos/metabolismo , Esfingosina/análogos & derivados , Esfingosina/metabolismoRESUMO
Hedgehog proteins constitute one of a small number of families of secreted signals that have a central role in the development of metazoans. Genetic analyses in flies, fish and mice have uncovered the major components of the pathway that transduces Hedgehog signals, and recent genome sequence projects have provided clues about its evolutionary origins. In this Review we provide an updated overview of the mechanisms and functions of this signalling pathway, highlighting the conserved and divergent features of the pathway, as well as some of the common themes in its deployment that have emerged from recent studies.
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Regulação da Expressão Gênica , Proteínas Hedgehog/metabolismo , Animais , Cílios/metabolismo , Drosophila melanogaster , Genoma , Proteínas Hedgehog/genética , Humanos , Ligantes , Lipídeos/química , Camundongos , Modelos Genéticos , Filogenia , Estrutura Terciária de Proteína , Transdução de Sinais , Fatores de Transcrição/metabolismoRESUMO
Nevoid basal cell carcinoma syndrome (NBCCS) is characterized by basal cell carcinoma, skeletal abnormalities, benign tumors including ovarian fibroma, and various other phenotypic expressions. Ovarian fibromas in NBCCS before puberty are very rare. We report a 6-year-old prepubescent girl with NBCCS showing skeletal abnormalities, medulloblastoma, and ovarian fibromas. The patient was referred to our hospital owing to abdominal distension. On admission, a huge elastic hard tumor was palpable and computed tomography showed a huge tumor of the left ovary. We performed a left salpingo-oophorectomy and diagnosed the tumor as a benign fibroma. Further examination of the computed tomography images showed skeletal abnormalities. In addition, the patient had a history of medulloblastoma at the age of 4 years. Therefore, we diagnosed NBCCS. A genetic examination indicated a novel 1 bp deletion in exon 18 (c.3055delG). Sequence analysis of exon 18 using DNA from the ovarian tumor revealed a mutant allele (c.3055delG) dominant to the wild-type allele, thus suggesting loss of heterozygosity in the PTCH1 gene, which is known to be associated with NBCCS. Conclusion On the basis of our experience, physicians treating pediatric ovarian tumors should be aware that such huge benign ovarian tumors may be a phenotype of NBCCS, as shown in our patient. In addition, genetic examination focusing on the PTCH1 gene might be important for diagnosis of NBCCS in pediatric patients.
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Síndrome do Nevo Basocelular/diagnóstico , Fibroma/diagnóstico , Neoplasias Ovarianas/diagnóstico , Criança , Diagnóstico Diferencial , Diagnóstico por Imagem , Feminino , Fibroma/cirurgia , Humanos , Japão , Neoplasias Ovarianas/cirurgiaRESUMO
Background/purpose: Vascular endothelial growth factor receptor (VEGFR) expression in oral squamous cell carcinoma (OSCC) promotes tumor growth through both autocrine and paracrine signaling. VEGF-positive OSCC cases are associated with a high depth of invasion, increased metastasis, and poor prognosis. In this study we established and then molecularly and functionally analyzed an OSCC cell line that co-expresses VEGF-A, VEGFR-1, and VEGFR-2, termed HCM-SqCC010 cells. Materials and methods: VEGF-A, VEGFR-1, and VEGFR-2 expression in HCM-SqCC010 cells were examined by immunohistochemistry and immunoblotting. Expression and inhibition of VEGF-A, VEGFR-1, and VEGFR-2 in HCM-SqCC010 cells were verified by quantitative real-time PCR. Results: Our analysis of HCM-SqCC010 cells revealed that their proliferation depended on VEGF-A, and selective inhibition of VEGFR-1 or VEGFR-2 resulted in decreased cell growth. Conclusion: We established an OSCC cell line, HCM-SqCC010, that expresses VEGF-A, VEGFR-1, and VEGFR-2. This triple-positive cell line showed no effect from a molecular targeted drug toward VEGF-A, but it did show strong cell growth inhibition in response to a VEGFR inhibitor. Thus, new therapeutic strategies against OSCC should include a VEGFR inhibitor.
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Mucoepidermoid carcinoma (MEC) is the most common malignant tumor of the major and minor salivary glands. Surgical resection is the only curative treatment and there is no effective post-operative therapy for MEC. The present study reports an Institutional Review Board-approved case of a 45-year-old Japanese female diagnosed with low-grade MEC in the hard palate. Radical resection, supraomohyoid neck dissection and antero-lateral thigh flap reconstruction was performed. A MEC cell line was then established from the resected tumor tissue. Short tandem repeat profiling confirmed the origin and authenticity of the cell line, that harbors a CRTC1-MAML2 translocation, which is frequently observed in MEC. Amphiregulin (AREG), identified as one of the targets of the CRTC1-MAML2 fusion gene, was expressed in the cell line. The AREG receptor, epidermal growth factor receptor (EGFR) was also highly phosphorylated. The results predicted that AREG-EGFR signaling, which is required for tumor growth and survival, might be activated in the cell line in a cell-autonomous manner. As AREG expression is associated with EGFR-targeted drug resistance, this cell line might assist with the identification of novel strategies for MEC treatment.
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Nevoid basal cell carcinoma syndrome (NBCCS), also known as Gorlin syndrome, is inherited in an autosomal dominant mode, and is characterized by a combination of developmental abnormalities and predisposition to form a variety of tumors. The hedgehog receptor Patched1 (PTCH1) has been identified as the gene mutated in NBCCS. We analyzed PTCH1 in two familial and three sporadic Japanese NBCCS cases, and identified five germline mutations in PTCH1. Two cases have a nonsense mutation (c.3058C>T and c.2760C>A), one a splice site mutation (c.584+2T>G), one a 1 bp insertion (c.2712_2713insA) and one a 1 bp deletion (c.980Gdel). All mutations induce truncation of the PTCH1 protein or could induce nonsense-mediated mRNA decay. The 11-year-old male patient with splice-site mutation (c.584+2T>G) had medulloblastoma (MB) at the age of 1 year. This is the first NBCCS patient with molecularly defined MB in Japan.
Assuntos
Síndrome do Nevo Basocelular/genética , Meduloblastoma/genética , Mutação/genética , Receptores de Superfície Celular/genética , Sequência de Bases , Criança , Primers do DNA/genética , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Receptores Patched , Receptor Patched-1 , Análise de Sequência de DNARESUMO
Hawaiian Drosophila offer an excellent model for adaptive evolution. More than 500 species are reported in Hawaiian islands, and there is considerable diversity in behavior and morphology. Such diversity is mainly driven by sexual selection. In this study qualitative and quantitative chemical compositions of cuticular hydrocarbons (CHCs) in 138 flies belonging to 27 Hawaiian Drosophila species, picture-winged and non picture-winged, were analyzed regarding sexual dimorphism, differences in saturation, branching position, and lengths of CHCs. We found significant variation in the CHC patterns. In several subgroups, new species show decreases in unsaturated hydrocarbons, and gradual increases in branched compounds, monomethylalkanes and dimethylalkanes, not commonly found in Drosophila. Moreover, branching positions gradually shifted towards internal carbons, and chain lengths increased in the new species. The long-term evolution of CHCs in the light of the recent evolutionary migration and adaptation history of Hawaiian Drosophila species along the developing archipelago was discussed.
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Alcanos/análise , Alcanos/química , Hidrocarbonetos/análise , Hidrocarbonetos/química , Alcanos/metabolismo , Animais , Evolução Biológica , Drosophila , Feminino , Havaí , Hidrocarbonetos/metabolismo , Tegumento Comum , Masculino , Preferência de Acasalamento Animal/fisiologia , Filogeografia , Especificidade da Espécie , Asas de AnimaisRESUMO
The spin gene was first identified by its mutant phenotype, which is characterized by extremely strong mate refusal by females in response to male courtship in Drosophila. Spin mutants are also known to be accompanied by a remarkable reduction in programmed cell death in the reproductive and nervous systems. To better understand the molecular functions of spin, we searched for its genetic modifiers. Forced expression of spin(+) in somatic cells as driven by ptc-Gal4 in the testis resulted in the invasion of mature sperm into the anterior testes tip, which is otherwise occupied only by immature germ cells. To obtain genes that modulate spin's effect, the gain-of-function spin phenotype was observed in the presence of a chromosome harboring an EP or GS P-element insertion, which initiates transcription of the genomic sequence neighboring the insertion site. We isolated th and emc as suppressors of spin and atg8a as a gene that reproduces the spin phenotype on its own. th encodes Inhibitor of apoptosis-1, and mammalian Id genes homologous to emc are known to inhibit apoptosis. atg8a encodes a protein essential for autophagy. These results suggest that spin promotes cell death mechanisms that are regulated negatively by th and emc and positively by atg8a.
Assuntos
Morte Celular/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/fisiologia , Proteínas de Membrana/metabolismo , Fenótipo , Animais , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Feminino , Masculino , Espermatogênese/genética , Testículo/metabolismoRESUMO
OBJECTIVES: The purpose of this study was to evaluate the fit, fracture load and aging resistance of the monolithic zirconia tooth-borne crowns with conventional and high-speed sintering. METHODS: The Y-TZP block was machined and sintered with conventional and high-speed sintering furnace. The marginal and internal gap between the crown and abutment was measured using a microscope and a fit checking material. A total of 28 crowns were further divided into an undegraded and a degraded group. An accelerated aging test was carried out on the degraded group. The crown was cemented and a fracture resistance was tested. X-ray diffraction was used to evaluate the crystalline structure. The data were analyzed with Student's t-test, and a one-way ANOVA and Tukey's multiple comparison test. RESULTS: There was no significant difference in mean marginal gap between the two groups. The mean internal gap was significantly greater in the speed sintering than in the conventional sintering (P <0.001). The mean fracture load of the conventional sintering crowns was not significantly different from that of speed sintering crowns after aging. The occurrence of monoclinic crystals of degraded crown was significantly higher than that of undegraded crown both in the conventional (P <0.001) and speed-sintering group (P <0.001). CONCLUSIONS: It was concluded that the monolithic zirconia crowns produced by high-speed sintering showed no significant difference in the marginal gap and the fracture load after aging compared to conventional sintering. Therefore, the high-speed sintering seems a valid method of producing tooth-borne monolithic zirconia crowns.
Assuntos
Desenho Assistido por Computador , Dente , Coroas , Planejamento de Prótese Dentária , Humanos , Teste de Materiais , ZircônioRESUMO
Basal cell nevus syndrome (BCNS or Gorlin syndrome, OMIM: 109400) is a rare autosomal dominant disorder with high penetrance. It is characterized by developmental anomalies and predisposition to tumors (for example, basal cell carcinoma (BCC) and medulloblastoma). PTCH1, the human homolog of the Drosophila patched gene, was identified as a gene responsible for BCNS. The PTCH1 protein is a Hedgehog (Hh) protein receptor and is pivotal for early development, stem cell maintenance and/or differentiation. We analyzed the six Japanese families with BCNS and identified six germline mutations in the PTCH1 gene. One family had a nonsense mutation (c.1196G>A), one had a 1-bp deletion (c.2029delA), two had 2-bp deletions (c.239_240delGA and c.1670_1671delCA) and one had a 58-bp duplication (c.1138_1195dup). They caused premature termination, resulting in the truncation of the PTCH1 protein. Analysis of a high-density single nucleotide polymorphism (SNP) mapping array showed a large approximately 1.2-Mb deletion, including the PTCH1 gene in one allele, in a family in which PTCH1 mutations were not identified at the sequence level. These data indicated that all the six families who were diagnosed with BCNS had mutations in the PTCH1 gene and that a single copy of a PTCH1 mutation causes BCNS.
Assuntos
Síndrome do Nevo Basocelular/genética , Mutação em Linhagem Germinativa/genética , Receptores de Superfície Celular/genética , Adulto , Povo Asiático/genética , Sequência de Bases , Criança , Pré-Escolar , Pontos de Quebra do Cromossomo , Feminino , Humanos , Japão , Masculino , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Receptores Patched , Receptor Patched-1 , Linhagem , Polimorfismo de Nucleotídeo Único/genéticaRESUMO
Human keratinocyte strains derived from the bulge region of plucked human follicles were successfully established from all 43 donors (age 24-76) regardless of the age and gender. The total cell number, number of population doublings and population doubling time were similar among the strains. These bulge-derived keratinocytes, BDKs, expressed keratin family genes specific to basal cell layers of the epidermis. They also expressed CD34, one of the bulge stem cell marker genes. The growth behavior and positivity of CD34 indicate that BDKs contain stem cells. BDKs were cultured until confluency or treated with CaCl2 to induce differentiation. Morphology and expression of keratin family genes in BDKs before and after differentiation induction with CaCl2 were similar to those of epidermal keratinocytes obtained from skin biopsies (NHEKs). However, expression levels of keratin-10, a prickle cell layer marker, in CaCl2-treated BDKs were lower than those in CaCl2-treated NHEKs. Higher expression of integrin-alpha6, a basal cell layer marker, was also noted in BDKs than in NHEKs after differentiation induction. Expression of stem cell marker genes other than CD34, including CD200, Sox2 and NANOG, was about the same at confluency in both cells, but significantly higher in BDKs than NHEKs after differentiation. These results indicate that BDKs were more refractory to differentiation than NHEKs. We then examined Wnt signaling inhibitor genes, DKK-3 and WIF-1 that function as tumor suppressors. DKK-3 expression decreased in both BDKs and NHEKs after CaCl2-induced differentiation. Expression of WIF-1 decreased 50% in BDKs one day after CaCl2 treatment and remained low, but was induced 1.7 times in NHEKs one day after CaCl2 treatment and further induced thereafter (>2.5 times), suggesting that WIF-1 may be involved in maintaining the differentiation-refractory status of BDKs. Since cancer stem cells in the skin have been reported to be similar to bulge-derived stem cells, our BDK strains may be of use in studying characteristics of cancer stem cells of the epidermis.
Assuntos
Diferenciação Celular/fisiologia , Folículo Piloso/citologia , Queratinócitos/fisiologia , Adulto , Células-Tronco Adultas/efeitos dos fármacos , Células-Tronco Adultas/fisiologia , Idoso , Biópsia , Cálcio/farmacologia , Técnicas de Cultura de Células/métodos , Proliferação de Células , Feminino , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/fisiologia , Perfilação da Expressão Gênica , Humanos , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Pele/patologia , Adulto JovemRESUMO
Cloninger's theory is that specific dimensions of temperament are associated with single neurotransmitter systems, and it is based on neurophysiologic and genetic approaches to the human traits. It suggests that overexpression of temperament could cause psychiatric illness. Based on this theory, we examined the correlation between reward dependence (RD) trait, measured with the Temperament and Character Inventory, and 5 polymorphisms in genes of norepinephrine pathways, ADRB1, COMT, PNMT, SLC18A1, and SLC6A2, in 85 Japanese female nursing students. We found that rs3764351 in PNMT was significantly associated with RD on Fisher's exact test (P = .029, P(corr) = .236). When haplotype analysis was performed for rs3764351 and rs876493 polymorphisms in the 5' flanking region of PNMT, 3 haplotypes were identified. Rs3764351 itself appeared to be correlated with RD in the present study of a specific population, although we could not demonstrate an association between RD and any of the haplotypes. Our findings have implications for the understanding of temperament using neurophysiologic approaches.
Assuntos
Povo Asiático/genética , Feniletanolamina N-Metiltransferase/genética , Recompensa , Temperamento , Adulto , Catecol O-Metiltransferase/genética , Feminino , Quinase 2 de Receptor Acoplado a Proteína G/genética , Frequência do Gene , Genótipo , Haplótipos , Humanos , Japão , Proteínas da Membrana Plasmática de Transporte de Norepinefrina/genética , Inventário de Personalidade , Polimorfismo de Nucleotídeo Único , Proteínas Vesiculares de Transporte de Monoamina/genéticaRESUMO
The fluorescence and physical properties of thulium-doped zirconia were investigated. A standard grade of zirconia (TZ-3Y-E) and two translucent dental zirconia materials (Zpex and Zpex Smile) were examined. The specimens were prepared by addition of 0-1.5 wt% Tm2O3 and then sintering. When exposed to UV light, the Tm2O3-doped zirconia exhibited blue fluorescence with a peak wavelength of 460 nm. The fluorescence intensity of Zpex and Zpex Smile was higher than that of TZ-3Y-E, with Zpex being more intense than Zpex Smile. Zpex exhibited maximum fluorescence intensity when doped with 0.8 wt% Tm2O3. XRD analysis revealed that TZ-3Y-E and Zpex contained primarily tetragonal zirconia, while Zpex Smile contained largely cubic phase zirconia. There were no changes observed in the microstructure or physical properties of the zirconia specimens when doped with Tm2O3.
Assuntos
Materiais Dentários/química , Fluorescência , Túlio/química , Zircônio/química , Dureza , Teste de Materiais , Microscopia Eletrônica de Varredura , Propriedades de Superfície , Raios Ultravioleta , Difração de Raios XRESUMO
Keratocystic odontogenic tumor (KCOT) is a benign tumor often associated with basal cell nevus syndrome (BCNS). Mutations in Patched 1 (PTCH1), the Hedgehog (Hh) receptor, are responsible for BCNS. BCNS is distinguished by morphological anomalies and predisposition to benign and malignant tumors, including medulloblastoma, basal cell carcinoma, KCOT and ovarian fibromas. Among these tumors, KCOT is the least well studied because a suitable model system is not available for its investigation. To enable KCOT to be studied, we established two KCOT cell lines, one from a BCNS case (designated as iKCOT1) and one from a sporadic KCOT case (designated as sKCOT1). The BCNSderived KCOT cell line, iKCOT1, retained a germline-mutated PTCH1 allele and a wild-type PTCH1 allele. The sporadic KCOT-derived KCOT cell line, sKCOT1, had different loss-of-function PTCH1 mutations on both alleles. Both cell lines expressed stem cell markers (CD44, SOX2 and BMI1), mesenchymal cell markers (CDH2, VIM and SNAI2) and a neurogenic marker (NEFL). Culture of the cell lines in high calcium concentration media induced expression of epithelial cell and keratinocyte marker proteins (CDH1, CLDN1, KRT10 and IVL). Parakeratosis, which is characteristic for KCOTs, was observed in 2-D cultures. The similarities in protein expression patterns between the two cell lines suggested that common mechanisms underlie the development of both types of KCOT and a probable common origin of KCOT cells.
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Síndrome do Nevo Basocelular/genética , Síndrome do Nevo Basocelular/patologia , Queratinócitos/patologia , Tumores Odontogênicos/genética , Tumores Odontogênicos/patologia , Adulto , Cálcio/farmacologia , Linhagem Celular Tumoral , Meios de Cultura , Feminino , Humanos , Receptor Patched-1/genética , Mutação Puntual , Células Tumorais CultivadasRESUMO
AIM: To investigate the possible role of polysaccharide-K (PSK) -related markers in predicting distant metastasis and in the clinical outcome of colorectal cancer (CRC). METHODS: Firstly, we used protein microarrays to analyze the in vitro expression profiles of potential PSK-related markers in the human colorectal adenocarcinoma cell line SW480, which carries a mutant p53 gene. Then, we investigated the clinical implications of these markers in the prognosis of CRC patients. RESULTS: ECA39, a direct target of c-Myc, was identified as a candidate protein affected by the anti-metastatic effects of PSK. Immunohistochemistry revealed that ECA39 was expressed at significantly higher levels in tumor tissues with distant metastases compared to those without (P<0.00001). Positive ECA39 expression was shown to be highly reliable for the prediction of distant metastases (sensitivity: 86.7%, specificity: 90%, positive predictive value: 86.7%, negative predictive value: 90%). A significantly higher cumulative 5-yr disease free survival rate was observed in the ECA39-negative patient group (77.3%) compared with the ECA39-positive patient group (25.8%) (P<0.05). CONCLUSION: Our results suggest that ECA39 is a dominant predictive factor for distant metastasis in patients with advanced CRC and that its suppression by PSK might represent a useful application of immunotherapy as part of a program of integrated medicine.
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Adenocarcinoma/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/metabolismo , Metástase Neoplásica/genética , Transaminases/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/patologia , Antibióticos Antineoplásicos/farmacologia , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Mutação/genética , Valor Preditivo dos Testes , Prognóstico , Proteoglicanas/farmacologia , Sensibilidade e Especificidade , Transaminases/genética , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismoRESUMO
The purpose of this study was to examine the translucency and low-temperature degradation of silica-doped experimental Y-TZP (Yttria-stabilized tetragonal zirconia polycrystal) containing almost no alumina. The experimental Y-TZP samples were sintered at either 1,450 or 1,500°C. The samples of commercially available translucent Y-TZP and conventional Y-TZP were used as controls. The contrast ratio (CR) and translucency parameter (TP) were obtained to compare the translucencies. In addition, the specimens were also subjected to an accelerated aging test. The results showed that the experimental Y-TZP sintered at 1,500°C and translucent Y-TZP exhibited almost the same level of translucency. During the accelerated aging test, the translucent Y-TZP underwent a substantial increase in monoclinic content, an index of degradation after the aging test. However, neither the experimental Y-TZP nor the conventional Y-TZP exhibited any appreciable change. It was concluded that the silica-doped Y-TZP will develop translucency and resistance to degradation when sintered at 1,500°C.
Assuntos
Dióxido de Silício , Zircônio , Teste de Materiais , Projetos Piloto , Propriedades de Superfície , Temperatura , ÍtrioRESUMO
OBJECTIVES: The objective of this study was to fabricate a radiopaque prosthetic fit-testing material, and to develop methodology to evaluate the fitting accuracy of prostheses three-dimensionally (3D) using a combination of the silicone replica technique and micro-computed tomography (µCT). METHODS: Eight types of prototype specimens of fit-testing materials were prepared by adding contrast agents (zirconia, alumina, and barium-glass) to a commercially available fit-testing material. These specimens were evaluated on their mechanical properties, X-ray absorption coefficients, reproducibility of cement space, and suitability for 3D analysis by µCT. Then, silicone replicas made from prototype specimens were assessed for accurate 3D morphology. Subsequently, color-mapping analyses of the silicone replicas were performed according to replica thickness, and the results were compared with stereomicroscopic images. RESULTS: The mechanical properties, X-ray absorption coefficients, and reproducibility of the cement space demonstrated that prototypes containing 20wt% zirconia (Zr-20) or barium glass (diameter 2µm; Ba2-20) were useful as fit-testing materials. However, the morphology of the Ba2-20 silicone replica was unable to be accurately described using µCT because of its low X-ray absorption threshold. Zr-20, however, could be clearly observed on µCT imaging. Furthermore, color-mapping analysis of the µCT images demonstrated that Zr-20 was the most suitable for 3D observation of prosthetic fit. SIGNIFICANCE: This method could allow any professional to evaluate the fit of any type of dental prosthesis, such as inlays, crowns, and fixed and removable dentures. This study demonstrated that the technique presented in the current study is able to accurately describe the abutment-crown prosthetic discrepancy based on silicone replicas.
Assuntos
Adaptação Marginal Dentária , Planejamento de Prótese Dentária , Microtomografia por Raio-X , Desenho Assistido por Computador , Coroas , Porcelana Dentária , Reprodutibilidade dos Testes , Propriedades de Superfície , ZircônioRESUMO
Somatic mutations of the BRCA1 associated protein-1 (BAP1) gene, which maps to 3p21, have been found in several tumors including malignant mesothelioma, uveal melanoma, and renal cell carcinoma (RCC). The role of BAP1 inactivation in tumor development remains unclear. It has been reported that Vhl knock-out mice did not develop RCC, but Vhl knock-out mice with single allele loss of Bap1 in nephron progenitor cells developed RCC, indicating that Bap1 inactivation may be essential in murine renal tumorigenesis. To clarify the role of BAP1 in human RCC development, we performed mutation analyses, including copy number detection of BAP1 and assessment of allelic imbalance using microsatellite polymorphisms on 3p, in 45 RCC samples derived from 45 patients without VHL or BAP1 germline mutation. Additionally, we analyzed the sequences of the VHL, PBRM1, and SETD2 genes, and examined promoter methylation of VHL. Using immunostaining, we also checked for expression of BAP1 protein, which is normally located in the nuclei. None of the RCCs had biallelic deletion of BAP1, but five (11.1%) showed a biallelic mutation (four with a sequence-level mutation with monoallelic loss and one with a biallelic sequence-level mutation); these cells were negative for nuclear BAP1 staining. These patients had worse recurrence-free survival than the patients without a biallelic mutation (p=0.046). However, there were no significant differences in worse outcome by multivariate analysis combined with age, T stage, histological subtype, infiltration and vascular invasion. In 35 RCCs (77.8%), monoallelic loss of BAP1 was accompanied by VHL biallelic mutation or VHL promoter hypermethylation. In five RCCs (11.1%), we detected 3p loss-of-heterozygosity, but the copy number of BAP1 was normal. Surprisingly, nuclear staining of BAP1 was negative in 10 out of 31 tumors (32.3%) with hemizygous normal BAP1, suggesting that haploinsufficiency may relate to RCC development.
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Carcinoma de Células Renais/patologia , Cromossomos Humanos Par 3/genética , Análise Mutacional de DNA/métodos , Neoplasias Renais/patologia , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , Ubiquitina Tiolesterase/genética , Ubiquitina Tiolesterase/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Haploinsuficiência , Humanos , Neoplasias Renais/genética , Neoplasias Renais/metabolismo , Masculino , Pessoa de Meia-Idade , Prognóstico , Análise de Sequência de DNA/métodos , Análise de SobrevidaRESUMO
Following the publication of this article, an interested reader drew to our attention an anomaly associated with the presentation of Figs. 2 and 3. Essentially, there was a direct duplication of certain of the western blotting data between Fig. 2 (the E-cadherin and Actin data) and Fig. 3C (the Zyxin and Actin data). After having re-examined our original data, we realize that we inadvertently duplicated the data from Fig. 2 in Fig. 3C (the Zyxin and Actin data). A corrected version of Fig. 3C (containing the true Zyxin and Actin data), and, by natural process, also of Fig. 6, are presented below, in which the Zyxin data in Figs. 3C and 6 are now correctly shown. Since the Zyxin data was a control for the siZyxin knockout of HOC313, this error did not affect the results of the Rho family analysis in this study. We sincerely apologize for this mistake, and thank the reader of our article who drew this matter to our attention. Furthermore, we regret any inconvenience this mistake has caused. [the original article was published in the International Journal of Oncology 42: 873-880, 2013; DOI: 10.3892/ijo.2013.1761].
RESUMO
Cisplatin (CDDP) is widely used to treat oral squamous cell carcinoma (OSCC), however, many patients exhibit acquired drug resistance. Yes-associated protein (YAP) is a transcriptional co-activator of the Hippo pathway that regulates organ size and promotes cell proliferation. YAP overexpression correlates with epithelial-mesenchymal transition and nodal metastasis, resulting in anti-tubulin drug resistance. Whether YAP overexpression is the cause of CDDP resistance in cancer cells is unclear, therefore, we investigated the correlation between YAP expression and CDDP sensitivity. We established three CDDP-resistant cell lines (OSC-19-R, SCCKN-R and HSC-3-R) from the OSCC parental cell lines. We also examined the expression levels of ATP7B, GST-π and ERCC1, which are strongly associated with CDDP resistance, and Hippo pathway-related proteins by western blotting. Using immunocytochemistry, we examined the cellular localization of YAP. Additionally, following knockdown of YAP using short interfering RNAs (siRNAs), we analyzed changes in sensitivity to CDDP. Compared with parental OSC-19 cells, OSC-19-R cells were obviously larger. Expression levels of YAP were not significantly different between OSC-19 and OSC-19-R. However, expression levels of phosphorylated YAP in OSC-19-R were decreased. We observed translocation of YAP from the cytoplasm to the nucleus in OSC-19-R cells. Knockdown of YAP using siRNAs revealed that sensitivity to CDDP was significantly increased. Translocation of YAP correlated with the acquisition of CDDP resistance. YAP could be a new therapeutic target for the treatment of patients with cancer that are resistant to CDDP.