ABSTRACT
Purpose This study sought to discern the clinical outcomes of intensity-modulated radiation therapy (IMRT) administered to the spine in patients who had undergone previous radiotherapy. Methods A total of 81 sites of 74 patients who underwent previous radiotherapy administered to the spine or peri-spine and subsequently received IMRT for the spine were analyzed in this study. The prescribed dose of 80 Gy in a biologically effective dose (BED) of α/β = 10 (BED10) was set as the planning target volume. The constraint for the spinal cord and cauda equine was D0.1 cc ≤ 100 Gy and ≤ 150 Gy of BED for re-irradiation alone and the total irradiation dose, respectively. Results The median follow-up period was 10.1 (0.992.1) months after re-irradiation, while the median interval from the last day of the previous radiotherapy to the time of re-irradiation was 15.6 (0.4210.1) months. Separately, the median prescript dose of re-irradiation was 78.0 (28.0104.9) of BED10. The median survival time in this study was 13.9 months, with 1-, 3-, and 5-year overall survival rates of 53.7%, 29.3%, and 26.6%, respectively. The 1-, 3-, and 5-year local control rates were 90.8%, 84.0%, and 84.0%, respectively. Neurotoxicity was observed in two of 72 treatments (2.8%) assessed after re-irradiation. Conclusion Re-irradiation for the spine using IMRT seems well-tolerated. Definitive re-irradiation can be a feasible treatment option in patients with the potential for a good prognosis (AU)
Subject(s)
Humans , Male , Female , Child , Adolescent , Young Adult , Adult , Middle Aged , Aged , Aged, 80 and over , Radiotherapy, Intensity-Modulated/adverse effects , Re-Irradiation/methods , Spinal Cord Neoplasms/radiotherapy , Survival Rate , Time Factors , Cauda Equina/radiation effects , Radiation Tolerance , Radiotherapy Dosage , Retrospective Studies , Spinal Cord Neoplasms/diagnostic imaging , Spinal Cord Neoplasms/mortalityABSTRACT
PURPOSE: This study sought to discern the clinical outcomes of intensity-modulated radiation therapy (IMRT) administered to the spine in patients who had undergone previous radiotherapy. METHODS: A total of 81 sites of 74 patients who underwent previous radiotherapy administered to the spine or peri-spine and subsequently received IMRT for the spine were analyzed in this study. The prescribed dose of 80 Gy in a biologically effective dose (BED) of α/ß = 10 (BED10) was set as the planning target volume. The constraint for the spinal cord and cauda equine was D0.1 cc ≤ 100 Gy and ≤ 150 Gy of BED for re-irradiation alone and the total irradiation dose, respectively. RESULTS: The median follow-up period was 10.1 (0.9-92.1) months after re-irradiation, while the median interval from the last day of the previous radiotherapy to the time of re-irradiation was 15.6 (0.4-210.1) months. Separately, the median prescript dose of re-irradiation was 78.0 (28.0-104.9) of BED10. The median survival time in this study was 13.9 months, with 1-, 3-, and 5-year overall survival rates of 53.7%, 29.3%, and 26.6%, respectively. The 1-, 3-, and 5-year local control rates were 90.8%, 84.0%, and 84.0%, respectively. Neurotoxicity was observed in two of 72 treatments (2.8%) assessed after re-irradiation. CONCLUSION: Re-irradiation for the spine using IMRT seems well-tolerated. Definitive re-irradiation can be a feasible treatment option in patients with the potential for a good prognosis.
Subject(s)
Radiotherapy, Intensity-Modulated , Re-Irradiation/methods , Spinal Neoplasms/radiotherapy , Adolescent , Adult , Aged , Aged, 80 and over , Analysis of Variance , Cauda Equina/radiation effects , Child , Female , Humans , Male , Middle Aged , Organs at Risk/radiation effects , Radiation Tolerance , Radiotherapy Dosage , Radiotherapy, Intensity-Modulated/adverse effects , Re-Irradiation/adverse effects , Relative Biological Effectiveness , Retrospective Studies , Spinal Cord/radiation effects , Spinal Neoplasms/diagnostic imaging , Spinal Neoplasms/mortality , Survival Rate , Time Factors , Young AdultABSTRACT
AIM: Glucagon-like peptide-1 is an incretin hormone secreted by the intestinal L-cell with a circadian rhythm that parallels expression of the core clock gene, Bmal1. Although feeding rats a high-fat/high-sucrose Western diet impairs rhythmic glucagon-like peptide-1 release, the mechanisms underlying this effect remain unclear. Therefore, the aim of this study was to determine the pathway(s) by which the saturated fat, palmitate, a major component of the Western diet, impairs circadian glucagon-like peptide-1 secretion. METHODS: Murine mGLUTag L-cells were synchronized, and the effects of palmitate pre-treatment on gene expression and glucagon-like peptide-1 secretion were determined, in addition to metabolite quantification, mitochondrial function analysis and enzyme inhibition and activation assays. Glucagon-like peptide-1 secretion was also analysed in ileal crypt cultures from control and Bmal1 knockout mice. RESULTS: Pre-treatment with palmitate dampened Bmal1 mRNA and protein expression and glucagon-like peptide-1 secretion at 8 but not 20 hours after cell synchronization (P < .05-.001). Glucagon-like peptide-1 release was also impaired in Bmal1 knockout cultures as compared to wild-type controls (P < .001). Palmitate pre-treatment reduced expression of the Bmal1 downstream target, nicotinamide phosphoribosyltransferase, the rate-limiting enzyme in the synthesis of NAD+ . This was paralleled by dampening of total NAD+ levels, as well as impaired mitochondrial function and ATP production (P < .05-.001). Whereas direct inhibition of nicotinamide phosphoribosyltransferase also decreased glucagon-like peptide-1 release, activation of this enzyme restored glucagon-like peptide-1 secretion in the presence of palmitate. CONCLUSION: Palmitate impairs L-cell clock function at the peak of Bmal1 gene expression, thereby impairing mitochondrial function and ultimately rhythmic glucagon-like peptide-1 secretion.
Subject(s)
ARNTL Transcription Factors/metabolism , Circadian Rhythm/drug effects , Enteroendocrine Cells/drug effects , Gene Expression Regulation/drug effects , Glucagon-Like Peptide 1/metabolism , Palmitates/pharmacology , ARNTL Transcription Factors/drug effects , Animals , Enteroendocrine Cells/metabolism , MiceABSTRACT
Exposures to metal toxicants in the environment disrupt normal physiological functions and have been linked to the development of a myriad of human diseases. While the molecular and cellular mechanisms underlying metal toxicities remain to be fully understood, it is well appreciated that metal toxicants induce cellular stresses and that how cells respond to the stresses plays an important role in metal toxicity. In this review, we focus on how metal exposures induce stresses in the endoplasmic reticulum (ER) to elicit the unfolded protein response (UPR). We document the emerging evidence that induction of ER stress and UPR in the development of human diseases is associated with metal exposures. We also discuss the role of the interplay between ER stress and oxidative stress in metal toxicity. Finally, we review recent advances in functional genomics approaches and discuss how applications of these new tools could help elucidate the molecular mechanisms underlying cellular stresses induced by environmental metal toxicants.
Subject(s)
Environmental Pollutants/toxicity , Metals/toxicity , Stress, Physiological/drug effects , Animals , Endoplasmic Reticulum Stress/drug effects , Genomics , Humans , Oxidative Stress/drug effects , Unfolded Protein Response/drug effectsABSTRACT
PURPOSE: The aim of this retrospective study was to evaluate the feasibility, safety, and effectiveness of stereotactic body radiotherapy (SBRT) for pulmonary metastases. PATIENTS AND METHODS: Between April 2007 and March 2011, 87 patients underwent SBRT for pulmonary metastases using the in-house Air-Bag System(TM) to obtain the four-dimensional image for treatment planning and to reduce intrafractional intrathoracic organ motion with abdominal compression to reduce the risk of radiation pneumonitis. Survival and respiratory adverse events were analyzed. RESULTS: The 2- and 3-year overall survival (OS) rates were 47 and 32 %, and the corresponding cause-specific survivals were 52 and 36 %. The 2- and 3-year OS rates were 57 and 49 % for patients in group 1, respectively, while the corresponding OS rates were 48 and 21 %, and 40 and 32 % for patients in groups 2 and 3, respectively. The 2- and 3-year local control (LC) rates were 80 and 80 %, respectively. The corresponding intrathoracic progression-free survival rates were 40 and 32 %, respectively. Concerning adverse respiratory events after SBRT for pulmonary metastases, 14 % were grade 0 (G0), 66 % G1, 13 % G2, 6 % G3, and 1 % G4. Concerning the adverse respiratory events (NCI-CTC) by grade scale, 1- and 2-year cumulative probabilities of radiation pneumonitis were 12 and 20 % for G2 and 4 and 10 % for G3/4, respectively. The mean values for cumulative V20 were 11.6 ± 8.5 %, 29.8 ± 18.6 %, and 25.7 ± 12.8 % in G0/1, G2, and G3/4, respectively. The number of pulmonary metastases that could be safely treated with SBRT was 6 PTVs (or seven gross tumor volumes) within a cumulative V20 of 30 % under the restricted intrafractional respiratory tumor motion using the Air-Bag System(TM). CONCLUSION: We propose that the number of pulmonary metastases that can be safely treated with SBRT is 6 PTVs with a cumulative V20 of 30 % under the restricted respiratory tumor motion using the Air-Bag System(TM). SBRT for pulmonary metastases offers locally effective treatment for recurrent or residual lesions after first line chemotherapy.
Subject(s)
Lung Neoplasms/secondary , Lung Neoplasms/surgery , Metastasectomy/instrumentation , Radiosurgery/instrumentation , Surgery, Computer-Assisted/instrumentation , Adult , Aged , Aged, 80 and over , Cause of Death , Combined Modality Therapy , Disease-Free Survival , Feasibility Studies , Female , Follow-Up Studies , Four-Dimensional Computed Tomography/instrumentation , Humans , Image Processing, Computer-Assisted/instrumentation , Immobilization/instrumentation , Lung Neoplasms/mortality , Magnetic Resonance Imaging/instrumentation , Male , Middle Aged , Neoplasm Recurrence, Local/surgery , Neoplasm, Residual/surgery , Patient Positioning/instrumentation , Postoperative Complications/etiology , Postoperative Complications/mortality , Radiation Pneumonitis/etiology , Radiation Pneumonitis/mortality , Radiotherapy Planning, Computer-Assisted/instrumentation , Retrospective Studies , Treatment OutcomeABSTRACT
Tumor necrosis factor-related apoptosis-inducing ligand-receptor 1 (TRAIL-R1) and tumor necrosis factor-related apoptosis-inducing ligand-receptor 2 (TRAIL-R2) are cell-surface receptors involved in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced cell-death signaling. TRAIL-R1 and TRAIL-R2 genes have recently been mapped to chromosome 8p21-22, which is a frequent site of allelic deletions in many types of human tumors, including non-Hodgkin's lymphoma (NHL). Because TRAIL/TRAIL receptor system plays an important role in lymphocyte homeostasis, we hypothesized that the mutations of TRAIL-R1 and TRAIL-R2 may be involved in the development of NHL and that such mutations may be responsible for the allelic losses of 8p21-22 in NHL. In this study, we analysed the entire coding region of TRAIL-R2 gene and the death domain region of TRAIL-R1 gene for the detection of the somatic mutations in a series of 117 human NHLs using polymerase chain reaction (PCR)-based single strand conformation polymorphism (SSCP) analysis. Overall, eight tumors (6.8%) were found to have two TRAIL-R1 gene mutations or six TRAIL-R2 gene mutations. Interestingly, of the eight mutations, six missense mutations (two TRAIL-R1 and four TRAIL-R2) were detected in the death domains and one nonsense mutation of TRAIL-R2 was detected just before the death domain. Our data suggest that somatic mutations of TRAIL-R1 and TRAIL-R2 genes may play a role in the pathogenesis of some NHLs and that TRAIL-R1 and TRAIL-R2 genes might be the relevant genes to the frequent loss of chromosome 8p21-22 in human NHL.
Subject(s)
Lymphoma, Non-Hodgkin/genetics , Mutation , Receptors, Tumor Necrosis Factor/genetics , Humans , Polymorphism, Genetic , Polymorphism, Single-Stranded Conformational , Receptors, TNF-Related Apoptosis-Inducing LigandABSTRACT
In this study, mutational and immunohistochemical analyses of beta-catenin were performed in 30 hepatoblastomas, to assess the prevalence of alterations of the Wnt pathway with respect to clinicopathological parameters and survival. Four missense mutations of beta-catenin (13.3%) were detected and there was strong immunoreactivity for beta-catenin in the cytoplasm and/or the nucleus in 97% of hepatoblastomas. Nuclear and cytoplasmic staining was demonstrated in 19 of 30 tumours (63%), while ten revealed only cytoplasmic staining. Statistically, this nuclear beta-catenin staining was significantly higher in the embryonal (Fisher exact test; p=0.00393) or undifferentiated type (p=0.00156) of hepatoblastoma than in the fetal type, but there was no difference between clinical stages I and II and clinical stages III and IV (p=0.175). Cumulative survival curves showed that nuclear beta-catenin staining (generalized Wilcoxon test; p=0.0088), undifferentiated histological type (p=0.0305), and clinical stages III and IV (p=0.0107) were significantly correlated with shorter survival time in these patients. Moreover, Cox multivariate analysis provides evidence that nuclear beta-catenin staining is the most important prognostic factor for survival (p=0.0090). It is therefore concluded that immunohistochemical analysis of beta-catenin might be a useful clinical tool for estimating the prognosis for patients with hepatoblastoma.
Subject(s)
Biomarkers, Tumor/metabolism , Cell Nucleus/metabolism , Cytoskeletal Proteins/metabolism , Hepatoblastoma/metabolism , Liver Neoplasms/metabolism , Trans-Activators , Biomarkers, Tumor/genetics , Cadherins/genetics , Cadherins/metabolism , Child , Child, Preschool , Cytoskeletal Proteins/genetics , Female , Follow-Up Studies , Genes, APC , Hepatoblastoma/genetics , Hepatoblastoma/pathology , Humans , Infant , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Loss of Heterozygosity , Male , Mutation , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Prognosis , Survival Rate , beta CateninABSTRACT
Many tumor cells, including hepatocellular carcinoma (HCC), express both Fas and its ligand on their surfaces, and it has remained a mystery why such cells do not spontaneously become apoptotic. In the current study, we analyzed the alterations of Fas structure and the expression of Fas and Fas ligand (FasL) and of Fas pathway inhibitors, including soluble Fas (sFas), Fas-associated phosphatase-1 (FAP-1), and bcl-2, in 50 cases of human HCC. Monoallelic loss of the Fas gene, as determined by loss of heterozygosity with intragenic polymorphisms, was observed in 5 of the 34 informative cases (15%), but none of the 50 cases showed Fas gene mutation. Expression of Fas and FasL was detected in 44 (88%) and 50 (100%) cases, respectively. sFas messenger RNA, as analyzed by in situ reverse-transcription polymerase chain reaction was expressed in 42 of the 50 cases (84%), and FAP-1 expression was observed in 40 of the 50 cases (80%). In contrast, none of the 50 cases showed bcl-2 expression. Our results showed that the majority of the HCCs (88%) coexpressed a death receptor, Fas and its cognate ligand, FasL, but all HCCs showed one or more alterations of the Fas pathway molecules known to inhibit Fas-mediated apoptosis. These findings suggest that the expression of sFas and FAP-1 and, in part, loss of Fas expression, rather than Fas gene alteration or bcl-2 expression, may be involved in the Fas resistance of HCC in vivo and that these mechanisms may play important roles in the pathogenesis of human HCC. HUM PATHOL 32:250-256.
Subject(s)
Carcinoma, Hepatocellular/chemistry , Liver Neoplasms/chemistry , Membrane Glycoproteins/analysis , fas Receptor/analysis , Adult , Aged , Apoptosis , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/pathology , Carrier Proteins/analysis , Fas Ligand Protein , Female , Gene Expression , Humans , Immunohistochemistry , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Male , Middle Aged , Mutation , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Protein Phosphatase 1 , Protein Tyrosine Phosphatase, Non-Receptor Type 13 , Protein Tyrosine Phosphatases/analysis , Proto-Oncogene Proteins c-bcl-2/analysis , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction , Solubility , fas Receptor/geneticsABSTRACT
It is now believed that genes regulating apoptosis are also important variables in cancer development. Fas, a transmembrane protein of the tumour necrosis factor receptor family, is a key molecule for cell death signalling. The mutation of the primary structure of the Fas gene might also be one of the possible mechanisms that disrupt Fas-mediated apoptosis in tumour cells. The purpose of this study was to determine whether somatic mutation of the Fas gene could be involved in the tumourigenesis of gastric cancer. Polymerase chain reaction (PCR)-based loss of heterozygosity (LOH) analysis with two intragenic polymorphic markers, and mutation analysis for the entire coding regions of the Fas gene were performed in 43 cases of gastric cancer, using PCR-single-strand conformational polymorphism sequencing. Five (11.6%) missense mutations were detected, only in the death domain of the Fas gene. Although these mutations were observed only in intestinal-type gastric cancers, there was no statistically significant difference in the frequency of Fas mutation between intestinal- and diffuse-type gastric cancer (p=0.068). Nine LOH out of 22 informative cases were also detected with one or both markers (41%). Three of them demonstrated a somatic mutation in the remaining allele, indicating the inactivation of both alleles. These results suggest that genetic alterations of the Fas gene may not only be limited to gastric cancer cell protection through Fas resistance, but may also play an important role in tumour promotion and/or progression in a subset of gastric cancer.
Subject(s)
Adenocarcinoma/genetics , Mutation, Missense/genetics , Stomach Neoplasms/genetics , fas Receptor/genetics , Adenocarcinoma/pathology , Adult , Aged , Aged, 80 and over , Alleles , Apoptosis/genetics , Female , Genetic Markers , Humans , Loss of Heterozygosity/genetics , Male , Middle Aged , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Stomach Neoplasms/pathologyABSTRACT
BACKGROUND & AIMS: There is increasing evidence that trefoil factor family 1 (TFF1) is a stabilizer of the mucous gel overlying the gastrointestinal mucosa that provides a physical barrier against various noxious agents. TFF1 knockout mice developed multiple gastric adenomas and carcinomas, suggesting that TFF1 is a gastric-specific tumor-suppressor gene. METHODS: We analyzed the somatic mutations and loss of heterozygosity (LOH) of the TFF1 gene using an intragenic polymorphic marker in 61 gastric tumors. The expression pattern of TFF1 was also examined by immunohistochemistry. RESULTS: We detected a total of 8 somatic mutations-1 (5.5%) of 18 adenomas and 7 (16.3%) of 43 carcinomas-that were all missense mutations confined to the loop I and loop II structure of TFF1. We detected LOH in 5 (1 in adenoma and 4 in cancer) of 30 (16.7%) informative gastric tumors with an intragenic polymorphic marker -2 base pairs (bp) upstream of the coding region of the TFF1 gene. Although 2 cases were noninformative, the 7 gastric cancers with mutation seemed to show the loss of the remaining allele except in 1 case, suggesting that TFF1 is a tumor-suppressor gene. We found loss of TFF1 expression in 44.2% of the gastric carcinomas, but there is no correlation between immunoreactivity and genetic alterations of the TFF1 gene. CONCLUSIONS: These results indicate that genetic alterations of TFF1 may lead to gastric mucosal barrier defects and contribute to the pathogenesis of gastric cancer.
Subject(s)
Growth Substances/genetics , Mucins , Multigene Family , Muscle Proteins , Mutation/genetics , Neuropeptides , Peptides/genetics , Stomach Neoplasms/genetics , Adult , Aged , Amino Acid Sequence/genetics , Base Sequence/genetics , DNA Mutational Analysis , Female , Gastric Mucosa/metabolism , Gastric Mucosa/pathology , Growth Substances/metabolism , Humans , Immunohistochemistry , Loss of Heterozygosity , Male , Middle Aged , Molecular Sequence Data , Peptides/metabolism , Reference Values , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathology , Trefoil Factor-2 , Trefoil Factor-3ABSTRACT
To determine the minimal region of deletion on 21q22 in gastric cancer, we performed a high-density loss of heterozygosity (LOH) study with eight polymorphic microsatellite markers. Among the 43 tumors examined, 20 (50%) of 40 informative carcinomas showed LOH at one or more loci. The peak LOH frequency was identified at D21S1820 (34.2%) in 21q22.3. This data suggests that this locus might harbor a new tumor suppressor gene in an area <0.332 Mb in physical map distance defined by D21S1820 and D21S49. Thus, we speculate that trefoil factor family 1 (TFF1), located in this narrow region, might be the most probable candidate gene involved in gastric cancer carcinogenesis.
Subject(s)
Adenocarcinoma/genetics , Chromosomes, Human, Pair 21/genetics , Loss of Heterozygosity , Stomach Neoplasms/genetics , Adenocarcinoma/pathology , Chromosome Banding , Chromosome Mapping , DNA, Neoplasm/genetics , Humans , Microsatellite Repeats , Stomach Neoplasms/pathologyABSTRACT
The SM1311 family is an Ashkenazi family with dominantly inherited predisposition to colorectal adenomas and carcinomas and has a high-penetrance locus in chromosome 15q, with a multipoint logarithm of the odds score of 3.06 at marker D15S118. In the present study, we performed a high-density loss of heterozygosity study with 13 polymorphic microsatellite markers, including D15S118, spanning 15q15.3-q22.1, on 70 cases of the sporadic form of colorectal tumors. Our deletion mapping data showed a locus at D15S968 in chromosomal sub-band 15q21.1 may harbor a tumor suppressor gene in an area <0.521 Mb in physical map distance defined by markers D15S514 and D15S222. THBS1, 0.185 Mb proximal to D15S968, is the nearest known gene to this specific narrow loss of heterozygosity region. Thus, we speculate that THBS1 might be the most probable candidate gene involved in colorectal cancer carcinogenesis.
Subject(s)
Adenocarcinoma/genetics , Adenoma/genetics , Chromosomes, Human, Pair 15/genetics , Colorectal Neoplasms/genetics , Genes, Tumor Suppressor/genetics , Loss of Heterozygosity , Chromosome Mapping , Disease Progression , Genetic Markers , Humans , Microsatellite Repeats/geneticsABSTRACT
Fas (Apo-1/CD95) is a cell-surface receptor involved in cell death signaling, and recent reports have suggested that defects within the Fas receptor pathway such as Fas mutation play an important part in the development and progression of human tumors. Burn scar-related squamous cell carcinoma of skin is a unique subtype of cutaneous squamous cell carcinoma, and tends to be more aggressive in nature than conventional squamous cell carcinoma. The molecular mechanisms underlying the development and progression of burn scar-related squamous cell carcinoma, however, are not clear. In this study, we analyzed the entire coding region and all splice sites of the Fas gene for the detection of the somatic mutations in a series of 50 conventional squamous cell carcinomas and 21 burn scar-related squamous cell carcinomas by polymerase chain reaction, single strand conformation polymorphism, and DNA sequencing. We detected mis-sense mutations in three of 21 burn scar-related squamous cell carcinomas (14.3%), whereas no mutation was detected in 50 conventional squamous cell carcinomas. Of the three Fas mutations detected in the burn scar-related squamous cell carcinomas, one was found in Fas ligand-binding domain, another one was identified in the death domain known to be involved in the transduction of an apoptotic signal, and the other one was found in the transmembrane domain. Our data show that some burn scar-related squamous cell carcinomas have Fas gene mutations in important regions for the apoptosis function and suggest that these mutations might be involved in the pathogenesis of burn scar-related squamous cell carcinomas. In addition, our results provide an important clue to understanding the difference between burn scar-related squamous cell carcinoma and conventional squamous cell carcinoma at the molecular level.
Subject(s)
Burns/complications , Carcinoma, Squamous Cell/genetics , Cicatrix/complications , Mutation , Skin Neoplasms/genetics , fas Receptor/genetics , Adolescent , Adult , Aged , Alleles , Base Sequence/genetics , Carcinoma, Squamous Cell/etiology , Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/pathology , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Mutation/genetics , Skin Neoplasms/etiology , Skin Neoplasms/metabolism , Skin Neoplasms/pathology , fas Receptor/metabolismABSTRACT
Hepatocyte growth factor/scatter factor (HGF/SF) and its receptor, Met, are known to play important roles in tumor cell migration, invasion, and metastasis. We analyzed the expression of these genes and the mutations in the kinase domain of the Met gene in 43 gastric carcinomas. Of the 43 cases, the Met and HGF/SF protein were expressed in 29 (67%) and 22 (51%), respectively. All of the cases with HGF/SF immunopositivity also expressed Met. Of 22 cases with HGF/SF immunopositivity, 13 (59%) expressed HGF/SF in tumor cells as well as fibroblasts. We detected no aberrant single-strand conformational polymorphism patterns, suggesting that there are no genetic alterations in the kinase domain of the Met gene. These results indicate that HGF/SF-mediated autocrine and/or paracrine stimulation and overexpression rather than structural alteration of its receptor may contribute to the development and progression of gastric carcinoma, and that expression of Met and HGF/SF may confer a growth advantage to neoplastic cells.
Subject(s)
Adenocarcinoma/enzymology , Mutation , Proto-Oncogene Proteins c-met/genetics , Stomach Neoplasms/enzymology , Adenocarcinoma/classification , Adenocarcinoma/pathology , Adenocarcinoma/physiopathology , Gene Expression , Hepatocyte Growth Factor/biosynthesis , Hepatocyte Growth Factor/genetics , Humans , Neoplasm Invasiveness , Polymorphism, Single-Stranded Conformational , Proto-Oncogene Proteins c-met/biosynthesis , Stomach Neoplasms/classification , Stomach Neoplasms/pathology , Stomach Neoplasms/physiopathologyABSTRACT
The Bcl10 gene, which encodes a protein with proapoptotic activity, recently has been identified on chromosome 1p22. In this study, we analyzed somatic mutations and deletions of the Bcl10 gene in a series of 439 tumor tissues from various histological origins that are known to have frequent loss of heterozygosity at chromosome 1p22. According to the LOH study at intragenic polymorphic sites, deletion of Bcl10 in informative cases was detected in 50% of malignant mesotheliomas, 33% of gastric carcinomas, 23% of breast carcinomas, 20% of hepatocellular carcinomas, 17% of lymphomas, 15% of colorectal carcinomas, 13% of laryngeal carcinomas, and 10% of male germ cell tumors (GCTs). In contrast, we detected Bcl10 mutations in 4 of 120 lymphomas (3.3%) and 2 of 78 GCTs (2.6%), respectively, but no mutation was found in the remaining solid tumors analyzed. Taken together, these data imply that Bcl10 may occasionally be involved in the pathogenesis of lymphoma and GCTs. However, the absence or low frequency of the mutation suggests that either Bcl10 is inactivated by other mechanisms or it is not the only target of chromosome 1p22 deletion in human tumors.
Subject(s)
Adaptor Proteins, Signal Transducing , Chromosomes, Human, Pair 1/genetics , Gene Deletion , Neoplasm Proteins/genetics , Neoplasms/genetics , Point Mutation/genetics , B-Cell CLL-Lymphoma 10 Protein , Breast Neoplasms/genetics , Carcinoma, Ductal, Breast/genetics , Carcinoma, Squamous Cell/genetics , Colorectal Neoplasms/genetics , Germinoma/genetics , Humans , Laryngeal Neoplasms/genetics , Lymphoma, B-Cell/genetics , Lymphoma, T-Cell/geneticsABSTRACT
Chromosome 8p21-22 is a frequent site of allelic deletions in many types of human tumors, including non-small cell lung cancer (NSCLC). Tumor necrosis factor-related apoptosis-inducing ligand-receptor 2 (TRAIL-R2) is a cell-surface receptor involved in cell death signaling. The TRAIL-R2 gene recently has been mapped to chromosome 8p21-22. To explore the possibility that the TRAIL-R2 gene might be the relevant gene to the frequent deletion of 8p21-22 in NSCLC, we have analyzed the entire coding region and all splice sites of TRAIL-R2 for the detection of the somatic mutations in a series of 104 NSCLCs. Overall, 11 tumors (10.6%) were found to have TRAIL-R2 gene mutations in the death domain known to be involved in the transduction of an apoptotic signal. Our data indicate that somatic mutation of TRAIL-R2 may play a role in the pathogenesis of some NSCLCs and that the TRAIL-R2 gene is one of the genes relevant to the frequent loss of chromosome 8p21-22 in NSCLC.
Subject(s)
Carcinoma, Non-Small-Cell Lung/genetics , Chromosomes, Human, Pair 8/genetics , Lung Neoplasms/genetics , Mutation/genetics , Receptors, Tumor Necrosis Factor/genetics , Adenocarcinoma/genetics , Amino Acid Sequence , Carcinoma, Squamous Cell/genetics , Humans , Loss of Heterozygosity , Molecular Sequence Data , Mutation, Missense/genetics , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Receptors, TNF-Related Apoptosis-Inducing LigandABSTRACT
Cross-linking of Fas and Fas ligand (FasL) induces apoptosis in Fas-bearing cells and regulates apoptosis. Fas is widely expressed in normal human tissues, but FasL expression has been considered to be restricted to lymphoid tissues. Recent studies have demonstrated that FasL is also expressed in some nonlymphoid tissues. To screen the in situ expression of FasL in normal human tissues, immunohistochemistry was performed using paraffin-embedded human tissues. FasL immunostaining was easily detected in testis, neurons, trophoblasts, tonsil, lymph node, Paneth cells, hepatocytes, renal tubular epithelium and bronchial epithelium, consistent with previous reports. Surprisingly, FasL was also expressed in many other cell types, including thymic medulla, skeletal muscle, cardiac muscle, pituitary gland, parathyroid gland, prostate glands, oocytes, epithelium of fallopian tube, endometrial glands, and gastric parietal cells. These findings demonstrate that FasL is widely expressed in human tissues and suggest that wide but cell-type specific expression of FasL may not only be implicated in the regulation of immune homeostasis but also in the regulation of cell death and life in many cell types in vivo.
Subject(s)
Membrane Glycoproteins/metabolism , Apoptosis/immunology , Fas Ligand Protein , Female , Homeostasis , Humans , Immunohistochemistry , Ligands , Lymphoid Tissue/immunology , Male , Tissue Distribution , fas Receptor/metabolismABSTRACT
The increased level of cytoplasmic beta-catenin through the mutations to either beta-catenin or adenomatous polyposis coli (APC) has been proposed as an important oncogenic step in various tumors. Gastric cancer showed frequent genetic alterations of the APC gene, and the risk for gastric cancer in familial adenomatosus polyposis patients is 10 times higher than that in the general population. These findings raise the possibility that mutations of beta-catenin may also be associated with the development of gastric cancer. We detected seven somatic mutations in a portion of exon 3 encoding for the glycogen synthase kinase 3beta phosphorylation consensus region of the beta-catenin gene in 43 gastric cancers. All of these mutations were missense mutations, of which five are in the highly conserved aspartic acid 32 and two are in serine 29; all of these seven mutations were detected exclusively in intestinal-type gastric cancers (7 of 26; 26.9%), but not in the diffuse-type (0 of 17). We concluded that disruption of the APC/beta-catenin/T cell factor-lymphoid enhancer binding factor pathway might play an important role especially in the development of intestinal-type gastric cancer.
Subject(s)
Cytoskeletal Proteins/genetics , Mutation, Missense , Stomach Neoplasms/genetics , Trans-Activators , Base Sequence , Genes, APC , Humans , Molecular Sequence Data , beta CateninABSTRACT
Fas (Apo-1/CD95) is a cell-surface receptor involved in cell death signaling. The key role of the Fas system in negative growth regulation has been studied mostly within the immune system, and somatic mutations of Fas gene in cancer patients have been described solely in lymphoid-lineage malignancies. However, many nonlymphoid tumor cells have been found to be resistant to Fas-mediated apoptosis, which suggests that Fas mutations, one of the possible mechanisms for Fas resistance, may be involved in the pathogenesis of nonlymphoid malignancies as well. In this study, we have analyzed the entire coding region and all splice sites of the Fas gene for the detection of the gene mutations in 44 human malignant melanomas in skin by polymerase chain reaction, single-strand conformation polymorphism, and DNA sequencing. Overall, 3 tumors (6.8%) were found to have the Fas mutations, which were all missense variants and identified in the cytoplasmic region (death domain) known to be involved in the transduction of an apoptotic signal. The data presented here suggest that somatic alterations of the Fas gene might lead to the loss of its apoptotic function and contribute to the pathogenesis of some human malignant melanomas.
Subject(s)
Melanoma/genetics , Skin Neoplasms/genetics , fas Receptor/genetics , Alleles , DNA Mutational Analysis , Heterozygote , Humans , Immunohistochemistry , Loss of Heterozygosity , Melanoma/metabolism , Mutation , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Skin Neoplasms/metabolism , fas Receptor/metabolismABSTRACT
Fas, a death receptor, is widely expressed in human tissue, but its expression, although a prerequisite for the induction of apoptosis, does not predict its biological function. To understand the mechanisms of Fas resistance in human tissues in vivo, we performed immunohistochemistry using an antibody against Fas-associated phosphatase-1 (FAP-1), which interacts with the cytosolic domain of Fas and inhibits Fas-mediated apoptosis. In normal human tissues, FAP-1 immunostaining was easily detected, for example, in renal tubules, skeletal muscle, myocardiocytes, pituitary gland, parathyroid gland, pancreatic islets, hepatocytes, testicular germ cells, prostatic glands, neurons, epithelium of fallopian tube, endometrial glands, trophoblasts, bronchial epithelial cells, and some types of gastrointestinal epithelial cells. In 123 (78%) of 158 cancers of various origins, including breast carcinomas, stomach carcinomas, colon carcinomas, lung carcinomas and several types of sarcomas, variable intensities of FAP-1 expression were evident. Taken together, these findings demonstrated that FAP-1 is widely expressed in normal human tissues and partly overlapped with Fas expression described in earlier reports, suggesting that FAP-1 may have an important role in the regulation of apoptosis in vivo. In addition, FAP-1 expression in cancers suggests that many cancers may be resistant to Fas-mediated apoptosis through the action of FAP-1 in vivo.