RESUMO
As an active constituent of the beetle Mylabris used in traditional Chinese medicine, cantharidin is a potent and selective inhibitor of protein phosphatase 2A (PP2A) that plays a crucial role in cell cycle progression, apoptosis, and cell fate. The role and possible mechanisms exerted by cantharidin in cell growth and metastasis of breast cancer were investigated in this study. Cantharidin was found to inhibit cell viability and clonogenic potential in a time- and dose-dependent manner. Cell cycle analysis revealed that cell percentage in G2/M phase decreased, whereas cells in S and G1 phases progressively accumulated with the increasing doses of cantharidin treatment. In a xenograft model of breast cancer, cantharidin inhibited tumor growth in a dose-dependent manner. Moreover, high doses of cantharidin treatment inhibited cell migration in wound and healing assay and downregulated protein levels of major matrix metalloproteinases (MMP)-2 and MMP-9. MDA-MB-231 cell migration and invasion were dose-dependently inhibited by cantharidin treatment. Interestingly, the members of the mitogen-activated protein kinase (MAPK) signaling family were less phosphorylated as the cantharidin dose increased. Cantharidin was hypothesized to exert its anticancer effect through the MAPK signaling pathway. The data of this study also highlighted the possibility of using PP2A as a therapeutic target for breast cancer treatment.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Cantaridina/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Animais , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Humanos , CamundongosRESUMO
As an active constituent of the beetle Mylabris used in traditional Chinese medicine, cantharidin is a potent and selective inhibitor of protein phosphatase 2A (PP2A) that plays a crucial role in cell cycle progression, apoptosis, and cell fate. The role and possible mechanisms exerted by cantharidin in cell growth and metastasis of breast cancer were investigated in this study. Cantharidin was found to inhibit cell viability and clonogenic potential in a time- and dose-dependent manner. Cell cycle analysis revealed that cell percentage in G2/M phase decreased, whereas cells in S and G1 phases progressively accumulated with the increasing doses of cantharidin treatment. In a xenograft model of breast cancer, cantharidin inhibited tumor growth in a dose-dependent manner. Moreover, high doses of cantharidin treatment inhibited cell migration in wound and healing assay and downregulated protein levels of major matrix metalloproteinases (MMP)-2 and MMP-9. MDA-MB-231 cell migration and invasion were dose-dependently inhibited by cantharidin treatment. Interestingly, the members of the mitogen-activated protein kinase (MAPK) signaling family were less phosphorylated as the cantharidin dose increased. Cantharidin was hypothesized to exert its anticancer effect through the MAPK signaling pathway. The data of this study also highlighted the possibility of using PP2A as a therapeutic target for breast cancer treatment.
Assuntos
Humanos , Animais , Feminino , Coelhos , Neoplasias da Mama/tratamento farmacológico , Cantaridina/farmacologia , Transdução de Sinais/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Antineoplásicos/farmacologia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Ensaios de Seleção de Medicamentos Antitumorais , Movimento Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacosRESUMO
A major obstacle in positional cloning is identifying the specific mutated gene from within a large physical contig. Here we describe the application of DNA microarray technology to a defined genomic region (physical map) to identify: (i) exons without a priori sequence data and (ii) the disease gene based on differential gene expression in a recessive disorder. The feasibility was tested using resources from the positional cloning of the Neimann-Pick Type C (NP-C) disease gene, NPC1. To identify NPC1 exons and optimize the technology, an array was generated from genomic fragments of the 110-kb bacterial artificial chromosome, 108N2, which encodes NPC1. First, as a test case for blindly identifying exons, fluorescently labeled NPC1 cDNA identified 108N2 fragments that contained NPC1 exons, many of which also contained intronic sequences and could be used to determine part of the NPC1 genomic structure. Second, to demonstrate that the NPC1 disease gene could be identified based upon differential gene expression, subarrays of 108N2 fragments were hybridized with fluorescently labeled cDNA probes generated from total RNA from hamster cell lines differentially expressing NPC1. A probe derived from the NP-C cell line CT60 did not detect NPC1 exons or other genomic fragments from 108N2. In contrast, several NPC1 exons were detected by a probe generated from the non-NP-C cell line 911D5A13, which was derived from CT60, and expressed NPC1 as a consequence of stable transduction with a YAC that contains NPC1 and encompasses 108N2. Thus, the array technology identified NPC1 as a candidate gene based on a physical contig and differential NPC1 expression between NP-C and non-NP-C cells. This technique should facilitate gene identification when a physical contig exists for a region of interest and mutations result in changes in the mRNA level of the disease gene or portions thereof.
Assuntos
Proteínas de Transporte , Clonagem Molecular/métodos , DNA/genética , Glicoproteínas de Membrana , Análise de Sequência com Séries de Oligonucleotídeos , Animais , Linhagem Celular , Cromossomos Bacterianos , DNA Complementar , Éxons , Perfilação da Expressão Gênica , Biblioteca Genômica , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Proteína C1 de Niemann-Pick , Hibridização de Ácido Nucleico , Proteínas/genéticaRESUMO
Tangier disease (TD) is an autosomal co-dominant disorder in which homozygotes have a marked deficiency of high density lipoprotein (HDL) cholesterol and, in some cases, peripheral neuropathy and premature coronary heart disease (CHD). Homozygotes are further characterized by cholesteryl ester deposition in various tissues throughout the body, most notably in those of the reticuloendothelial system. Several studies have demonstrated that the excess lipid deposition in TD is due to defective apolipoprotein-mediated efflux of cellular cholesterol and phospholipids. Although much progress has been made in our understanding of the metabolic basis of TD, the precise molecular defect had remained elusive until very recently. By positional cloning methods, we: 1) confirm the assignment of TD to chromosome 9q31, 2) provide evidence that human ATP-binding cassette-1 (hABC-1) maps to a 250 kb region on 9q31, and 3) describe novel deletion, insertion, and missense mutations in the gene encoding hABC-1 in four unrelated TD kindreds. These results establish a causal role for mutations in hABC-1 in TD and indicate that this transporter has a critical function in the regulation of intracellular lipid trafficking that dramatically affects plasma HDL cholesterol levels.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Mutação , Doença de Tangier/genética , Sequência de Bases , Cromossomos Humanos Par 9 , Primers do DNA , Feminino , Heterozigoto , Homozigoto , Humanos , Masculino , LinhagemRESUMO
OBJECTIVE: In order to search a safer argon laser photodynamic method, this study used 488 nm wavelength and common mixed spectrum argon laser PDT for treating Port Wine Stain. The therapeutic effects and side effect were compared. METHODS: Fifty two cases of PWS were divided into two groups randomly. Argon laser PDT by two ways of laser irradiation were used. RESULTS: Our data revealed 19.23% cure rate in 488 nm laser PDT group, while that of mixed wavelength laser group was 11.54%. There was no significant difference in the two groups. Also the effective rate of 488 nm laser group was 92.31% and that of mixed group was 88.46%. But the difference was not significant statistically. Side effect occurred often in mixed spectrum laser group. That was 57.69% for mixed group, 38.46% for 488 nm group. CONCLUSION: Argon laser PDT at 488 nm had the similar therapeutic result contrasted to mixed spectrum laser group, but it was safe.
RESUMO
OBJECTIVE:There were 13 cases underwent operation.24 cases underwent cryosurgery only and 70 cases received comprehensive management,which as cryosurgery,operation,chemotherapy and immuno-therapy.RESULTS:The 3'and 5' year survival rate of three groups were 0.0%,0.0%,37.50%,31.25% and 57.14%,36.07%.There are significant differences between operation group and cryosurgery group and between operation group and comprehensive treatment group.Although the third group got the better result,but the difference between late two groups was not statistical significant.CONCLUSION:It is suggested that comprehensive treatment be the routine regime of melanoma in head and neck. Our experience with review of literature covered the classification, surgical principle and complication was presented.
RESUMO
Niemann-Pick disease type C (NP-C) is an autosomal recessive lipidosis linked to chromosome 18q11-12, characterized by lysosomal accumulation of unesterified cholesterol and delayed induction of cholesterol-mediated homeostatic responses. This cellular phenotype is identifiable cytologically by filipin staining and biochemically by measurement of low-density lipoprotein-derived cholesterol esterification. The mutant Chinese hamster ovary cell line (CT60), which displays the NP-C cellular phenotype, was used as the recipient for a complementation assay after somatic cell fusions with normal and NP-C murine cells suggested that this Chinese hamster ovary cell line carries an alteration(s) in the hamster homolog(s) of NP-C. To narrow rapidly the candidate interval for NP-C, three overlapping yeast artificial chromosomes (YACs) spanning the 1 centimorgan human NP-C interval were introduced stably into CT60 cells and analyzed for correction of the cellular phenotype. Only YAC 911D5 complemented the NP-C phenotype, as evidenced by cytological and biochemical analyses, whereas no complementation was obtained from the other two YACs within the interval or from a YAC derived from chromosome 7. Fluorescent in situ hybridization indicated that YAC 911D5 was integrated at a single site per CT60 genome. These data substantially narrow the NP-C critical interval and should greatly simplify the identification of the gene responsible in mouse and man. This is the first demonstration of YAC complementation as a valuable adjunct strategy for positional cloning of a human gene.
Assuntos
Cromossomos Artificiais de Levedura/genética , Cromossomos Humanos Par 18 , Cromossomos Humanos Par 7 , Doenças de Niemann-Pick/genética , Animais , Células CHO , Mapeamento Cromossômico , Clonagem Molecular , Cricetinae , DNA Complementar , Humanos , CamundongosRESUMO
An integrated human-mouse positional candidate approach was used to identify the gene responsible for the phenotypes observed in a mouse model of Niemann-Pick type C (NP-C) disease. The predicted murine NPC1 protein has sequence homology to the putative transmembrane domains of the Hedgehog signaling molecule Patched, to the cholesterol-sensing regions of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and SREBP cleavage-activating protein (SCAP), and to the NPC1 orthologs identified in human, the nematode Caenorhabditis elegans, and the yeast Saccharomyces cerevisiae. The mouse model may provide an important resource for studying the role of NPC1 in cholesterol homeostasis and neurodegeneration and for assessing the efficacy of new drugs for NP-C disease.
Assuntos
Colesterol/metabolismo , Modelos Animais de Doenças , Doenças de Niemann-Pick/genética , Proteínas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Homeostase , Humanos , Hidroximetilglutaril-CoA Redutases/química , Peptídeos e Proteínas de Sinalização Intracelular , Lisossomos/metabolismo , Proteínas de Membrana/química , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Dados de Sequência Molecular , Mutação , Proteína C1 de Niemann-Pick , Doenças de Niemann-Pick/metabolismo , Fenótipo , Sinais Direcionadores de Proteínas/química , Proteínas/química , Proteínas/fisiologia , Homologia de Sequência de AminoácidosRESUMO
To identify candidate genes for Branchio-oto-renal (BOR) syndrome, we have made use of a set of cosmids that map to 8q13.3, which has previously been shown to be involved in this syndrome. These cosmids were used as genomic clones in the attempts to isolate corresponding cDNAs using a modified hybrid selection technique. cDNAs from the region were identified and used to search for sequence similarity in human or other species. One cDNA clone was found to have 89% sequence similarity to the bovine B22 subunit of NADH-ubiquinone oxidoreductase, a mitochondrial protein in the respiratory electron transport chain. Given the history of other mitochondrial mutations being involved in hearing loss syndromes, this gene should be considered a strong candidate for involvement in BOR.
Assuntos
Anormalidades Múltiplas/genética , Cromossomos Humanos Par 8/genética , NADH NADPH Oxirredutases/genética , Anormalidades Múltiplas/enzimologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Região Branquial/anormalidades , Bovinos , Cosmídeos/genética , DNA Complementar/genética , Surdez/genética , Transporte de Elétrons/genética , Complexo I de Transporte de Elétrons , Genes , Humanos , Rim/anormalidades , Mitocôndrias/enzimologia , Dados de Sequência Molecular , NADH NADPH Oxirredutases/química , Alinhamento de Sequência , Homologia de Sequência , SíndromeRESUMO
Genetic linkage analysis has previously mapped the locus for the autosomal dominant disorder branchio-oto-renal syndrome (BOR) to the pericentric region of chromosome 8q. A YAC contig spanning the putative BOR region, from D8S543 to D8S541, was constructed and confirmed by sequence-tagged site content mapping using microsatellite markers and by DNA hybridization analysis. YACs spanning the BOR interval were used as fluorescence in situ hybridization probes on a cell line from a patient with BO and tricho-rhino-phalangeal syndrome I that involves a chromosome 8q rearrangement. In addition to the cytogenetically defined direct insertion of material from 8q13.3-q21.13 into 8q24.11, a previously unidentified deletion of just under one megabase was found in 8q13.3. These data narrowed the most likely location of the BOR gene to a region corresponding to the proximal two-thirds of YAC 869E10 between D8S543 and D8S279.
Assuntos
Anormalidades Múltiplas/genética , Região Branquial/anormalidades , Transtornos da Audição/genética , Rim/anormalidades , Síndrome de Langer-Giedion/genética , Sequência de Bases , Mapeamento Cromossômico , Cromossomos Artificiais de Levedura/genética , Clonagem Molecular , Primers do DNA , Humanos , Dados de Sequência Molecular , Deleção de SequênciaRESUMO
Malignant melanoma is a kind of tumor with higher malignant and lower therapeutic effect.it is demonstrated by experiment that melanoma cells have antigenicity and refer to immunology.BCG is a kind of biologic response modifier(BRM).it is able to promote physical antitumor ability.There were 42 patients with oral and maxillofacial malignant melanoma who received freezing therapy and surgery in this paper.11 cases of them added BCG immunotherapy.The result is that the median survival time of therapy plus BCG group is 4 years,and 3 years,5 years,7 years survival rate are 89%,72% and 32%,over 8 years survival rate is 18%.The median survival time of another group is 2 years,and 3 years,5 years,7 years survival rate are 55%,24% and 12%,over 8 years survival rate is 3%.There is a statistically significant difference between these two groups and BCG group has a longer survival time.This preliminary study demonstrated that BCG adjuvant therapy could decrease recurrence of metastases and increase patients survival time.
RESUMO
Wedge biopsies of liver from 155 patients with advanced schistosomiasis japonica were observed pathologically, and HBsAg and HBcAg in liver were tested with double bridge peroxidase-anti-peroxidase (PAP) method. 88 of 155 cases (56.8%) were found to be HBsAg and/or HBcAg (HBAg) positive in liver. Eosinophilic intranuclear inclusions were observed in the hepatocytes of 30 cases (19.4%), in which 18 (60%) were also HBAg positive in liver. These inclusions were considered to be the markers of several virus infections, such as cytomegalovirus, herpes simplex virus, etc. The patients with positive HBAg and/or inclusion in liver had significantly more severe pathological changes in liver parenchyma. The results indicate that in addition to hepatitis B, complication with other viral infections in liver, which produce eosinophilic intranuclear inclusion, may also aggravate the pathological changes in liver and may be one of the causes of portal cirrhosis in patients with advanced schistosomiasis japonica (Fig. 1). schistosomiasis japonica