ABSTRACT
BACKGROUND: Stat3 (Signal transducer and activator of transcription-3) is an oncogene that plays a critical role in regulating fundamental processes associated with malignant transformation and cell survival. It participates in oncogenesis through upregulation of genes encoding apoptosis inhibitors (Bcl-xL) and cell cycle regulators (cyclin D1). The expression of Stat3, Bcl-xL and cyclin D1 protein has not been investigated in extramammary Paget disease (EMPD). OBJECTIVES: To study the expression of phosphorylated Stat3 (p-Stat3), Bcl-xL and cyclin D1 protein in EMPD and to evaluate the relationships among them. METHODS: Thirty-six tissue samples from 34 patients with primary EMPD were subjected to immunohistochemical staining for p-Stat3, cyclin D1 and Bcl-xL. RESULTS: Thirty-five of 36 specimens were clearly positive for p-Stat3 in EMPD, while 30 of 36 and 32 of 36 were positive for cyclin D1 and Bcl-xL expression, respectively. In all of four invasive EMPD specimens, strong and frequent expression of these three molecules was evident; moreover, two invasive EMPD specimens with lymph nodal metastasis showed very strong nuclear and membranous p-Stat3 staining. Two metastatic lymph node specimens showed very strong nuclear and local membrane p-Stat3 staining. There were significant correlations between p-Stat3 and cyclin D1 expression and between p-Stat3 and Bcl-xL expression. CONCLUSIONS: Our study shows that the expression of p-Stat3, cyclin D1 and Bcl-xL may play a pivotal role in the pathogenesis of EMPD.
Subject(s)
Neoplasm Proteins/metabolism , Paget Disease, Extramammary/metabolism , STAT3 Transcription Factor/metabolism , Adult , Aged , Aged, 80 and over , Cyclin D1/metabolism , Female , Humans , Immunoenzyme Techniques , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Invasiveness , Paget Disease, Extramammary/pathology , Paget Disease, Extramammary/secondary , Phosphorylation , Skin/metabolism , bcl-X Protein/metabolismABSTRACT
The MHC-DRB1 gene is known to display the most extensive allelic polymorphisms among MHC class II genes. We attempted the selective identification of chimpanzee (Pan troglodytes) DRB1 (Patr-DRB1) alleles using the polymerase chain reaction (PCR) technique in three steps: first, we performed Patr-DRB1*02 lineage-specific 8-kb PCR for *02 lineage detection in each chimpanzee; second, we performed 620-bp PCR for amplification of full-length exon 2; and finally, we carried out an insert check using the pattern of microsatellite repeat length variability. In the genomic DNA of 23 chimpanzees, nine Patr-DRB1 alleles containing two new alleles were detected. Our approach provides a relatively effective method of identifying Patr-DRB1 alleles in individual chimpanzees and should also contribute to our understanding of the features of MHC molecules in non-human primates.
Subject(s)
Alleles , Genetic Variation , HLA-DR Antigens/genetics , Pan troglodytes/genetics , Polymorphism, Genetic , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Genes, MHC Class II , HLA-DRB1 Chains , Microsatellite Repeats , Molecular Sequence Data , Pan troglodytes/immunology , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Species SpecificitySubject(s)
Bone Marrow Cells/cytology , Dendritic Cells/cytology , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology , Lymphocytes/immunology , Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/physiology , Animals , Bone Marrow Cells/drug effects , Bone Marrow Cells/immunology , Culture Media, Conditioned , Dendritic Cells/drug effects , Dendritic Cells/immunology , Humans , Immunophenotyping , Interleukin-6/pharmacology , Lymphocyte Activation , Mice , Mice, Knockout , Proto-Oncogene Proteins/physiology , Rats , Rats, Inbred Lew , Receptor Protein-Tyrosine Kinases/physiology , Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/deficiency , Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Recombinant Proteins/pharmacology , Spleen/immunology , Tumor Necrosis Factor-alpha/pharmacology , fms-Like Tyrosine Kinase 3ABSTRACT
The chemosensitivity test with growth chamber (GC), a semi permeable polymer matrix, was conducted using human tumor xenografts, comparing the results with those of in vivo nude mouse system. Xenografts used were MX-1, St-4, Co-3, and Co-4. Normal stromal cells, SM-74, a cell line derived from human adult skin fibroblast, and Clone-A-31, a cell line from BALB/c nu/nu nude mice were used as the control. Dissociated tumor cell suspension in 200 microliters of Medium 199 was plated into GC (80,000 cells, chamber) and incubated with a various concentration of mitomycin C(MMC), cisplatin (DDP), 5-fluorouracil (5-FU), and adriamycin (ADM) at the various concentrations. After incubation of 1 wk, the activity of hexosaminidase was measured with ELISA assay using p-nitrophenyl-N-acetyl-glucosaminide. The antitumor activity of the agents against human tumor xenografts was dose dependent, and the antitumor spectra obtained by GC assay was essentially identical to in vivo nude mouse system. On the other hand, no evaluable optical density could be obtained with normal stromal cells, SM-74 and Clone-A-31. The optimal cutoff concentration of each drugs to predict the in vivo results was estimated to be 10 micrograms/ml for MMC, 15 micrograms/ml for DDP and 5-FU, and 0.7 microgram/ml for ADM. The predictability of GC assay was 77%, including 89% sensitivity and 70% specificity. Since GC assay could eliminate the normal stromal cells because of the characteristic of chamber surface, this assay was thought to be useful for the clinical chemosensitivity test of human cancers containing a large number of normal stromal cells.
