RESUMEN
PURPOSE: A novel human gene, designated HRad17, was identified as the human homologue of the Rad17 of Schizosaccharomyces pombe and Rad24 of Saccharomyces cerevisiae. In yeast, these genes play a critical role in maintaining genomic stability. The aim of this study was to evaluate the expression of HRad17 in human breast cancer. EXPERIMENTAL DESIGN: We investigated HRad17 mRNA expression in 64 cases of human breast cancer by means of reverse-transcription-PCR, in situ hybridization, and immunohistochemistry. RESULTS: The HRad17 mRNA was overexpressed in 35 cases (54.7%). Twenty-four (68.6%) of 35 cases with HRad17 overexpression in cancer tissues were node-positive, whereas only 8 (27.6%) of 29 cases without HRad17 overexpressions were node-positive. The expression of HRad17 mRNA correlated with both lymph node metastasis (P = 0.001) and high Ki67 labeling index (P = 0.006). Although not significantly different, expression of HRad17 mRNA tended to correlate with tumor size (P = 0.06) and expression of mutant p53 protein (P = 0.10). Furthermore, expression of HRad17 mRNA was an independent predictor of axillary lymph node metastasis as well as of lymphatic permeation by multivariate analysis (P < 0.0001). CONCLUSIONS: Our study demonstrates that HRad17 might be related to the development of lymph node metastasis in human breast cancers. Although its function still remains unclear, the expression of HRad17 mRNA could open up a new window for the diagnostic staging and treatment of human breast cancers.
Asunto(s)
Neoplasias de la Mama/genética , Proteínas de Ciclo Celular/genética , ARN Mensajero/metabolismo , Adulto , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Proteínas de Ciclo Celular/análisis , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Inmunohistoquímica , Hibridación in Situ , Antígeno Ki-67/análisis , Metástasis Linfática/genética , Metástasis Linfática/patología , Persona de Mediana Edad , Análisis Multivariante , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales Cultivadas , Proteína p53 Supresora de Tumor/análisis , Proteína p53 Supresora de Tumor/genéticaRESUMEN
Radiotherapy is a useful component of treatment for esophageal cancer. Identification of the genes that are differentially expressed between radiosensitive and radioresistant cancer cells is important for predicting clinical effectiveness of radiotherapy. We established human esophageal cancer cell lines resistant to X-ray. Using differential display, we obtained one gene that was expressed in radiosensitive cells but was rarely expressed in radioresistant cells, and that gene was identical with hepatoma-derived growth factor (HDGF), an acidic polypeptide with mitogenic activity for fibroblasts. The semiquantitative reverse transcription-PCR assay confirmed that HDGF mRNA expression was reduced in established radioresistant cells, and its reduction was associated with reduced sensitivity to irradiation. Radiotherapy was more effective in clinical cases with high HDGF mRNA expression compared with cases with low expression (P < 0.05). The findings demonstrate that HDGF may play an important role in radiosensitivity, and it could be a novel marker predicting effectiveness of radiotherapy in clinical cases.
Asunto(s)
Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/radioterapia , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/radioterapia , Sustancias de Crecimiento/genética , Péptidos y Proteínas de Señalización Intercelular , Tolerancia a Radiación/genética , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma de Células Escamosas/metabolismo , Neoplasias Esofágicas/metabolismo , Femenino , Factor 2 de Crecimiento de Fibroblastos/biosíntesis , Factor 2 de Crecimiento de Fibroblastos/genética , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Sustancias de Crecimiento/biosíntesis , Humanos , Masculino , Persona de Mediana Edad , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Células Tumorales Cultivadas/efectos de la radiaciónRESUMEN
BACKGROUND: Recurrent and metastatic carcinoma of the colorectum remains a major problem. This may be ascribed to the presence of micrometastasis at diagnosis. The purpose of this study was to analyze prospectively the clinical value of detecting K-ras mutations in the perioperative circulating blood from patients with colorectal carcinoma. METHODS: Twenty-four patients whose tumor carried mutations in codon 12 of the K-ras gene were studied for the presence of cancer cells in perioperative blood samples, in particular, tumor drainage samples. A detection assay using CD45 immunomagnetic separation plus nested mutant allele specific amplification (MASA) was performed. RESULTS: K-ras mutations in CD45 negative cells in tumor drainage blood were detected in 7 (29.2%) of 24 patients. There was no significant relationship between the presence of a K-ras mutation and clinicopathological features. Four (57.1%) of the seven patients with a positive K-ras mutation in drainage blood had early recurrent disease. Of the 17 patients with no K-ras mutation, none developed metastatic disease. The recurrence rate of the K-ras mutation positive group was higher than that of the K-ras mutation negative group (P < .01). There was a significant difference, regarding prognosis, between K-ras mutation positive and negative groups (P < .01). CONCLUSIONS: This preliminary study demonstrates that the detection of circulating cancer cells in the tumor drainage blood by our new assay system may provide a predictor of recurrence and metastasis of colorectal cancer.
Asunto(s)
Adenocarcinoma/sangre , Adenocarcinoma/genética , Neoplasias Colorrectales/sangre , Neoplasias Colorrectales/genética , Genes ras/genética , Mutación/genética , Adenocarcinoma/patología , Anciano , Neoplasias Colorrectales/patología , Drenaje , Femenino , Humanos , Periodo Intraoperatorio , Antígenos Comunes de Leucocito/sangre , Masculino , Persona de Mediana Edad , Células Neoplásicas Circulantes/patología , Análisis de Supervivencia , Resultado del TratamientoRESUMEN
Expression of the chemokine stromal cell-derived factor-1alpha (SDF-1alpha) is absent from many carcinomas, including hepatomas. We note an early signalling defect in the hepatocellular carcinoma (HCC) cell line HepG2 that expresses the CXCR4 receptor and binds biotin-labelled SDF, but fails to stimulate downstream signalling events after engagement with SDF. In HepG2, the SDF/CXCR4 interaction did not result in calcium influx, phosphorylation and internalization of CXCR4, nor in a rapid phosphorylation of p44/42 MAP kinase. There were no CXCR4 mutations in the second chemokine binding loop or C terminal phosphorylation and internalization domains. The downstream signalling machinery in HepG2 appears to be intact since transfection of wild-type CXCR4 restored functional responsiveness. We conclude that HepG2 is unresponsive to SDF stimulation because of a defect located after receptor binding but before the activation of the signalling cascade. A hypothetical blocking molecule could hinder receptor internalization or CXCR4 signalling.
Asunto(s)
Carcinoma Hepatocelular , Quimiocinas CXC/metabolismo , Neoplasias Hepáticas , Sistema de Señalización de MAP Quinasas/fisiología , Receptores CXCR4/metabolismo , Quimiocina CXCL12 , Regulación hacia Abajo/fisiología , Citometría de Flujo , Regulación Neoplásica de la Expresión Génica , Humanos , Células Jurkat , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Fosforilación , Receptores CXCR4/genética , Análisis de Secuencia de ADN , TransfecciónRESUMEN
Recently, mammalian heparanase was cloned, and its probable function in tumor progression was reported. However, its expression in human clinical cancers has not been fully studied. Thus we determined the heparanase mRNA expression in 30 esophageal cancer cell lines and 144 clinical samples including 38 esophageal squamous cell carcinomas, 71 gastric adenocarcinomas, and 35 colorectal adenocarcinomas. The fresh surgical specimens of cancer tissue (T) and its paired normal tissue (N) were used. The heparanase mRNA was evaluated by reverse transcriptase-polymerase chain reaction, and the T/N expression ratio was determined in clinical cases. All 30 esophageal cancer cell lines expressed heparanase mRNA. The T/N ratio was determined as high (> or =1.3), equal (0.8 approximately 1.2) or low (< or = 0.7) in each clinical case. In cases of esophageal cancer, 7 showed high, 10 equal and 21 low expression. In cases of colorectal cancer, 3 showed high, 16 equal and 16 low expression. On the other hand, 42 showed high, 22 equal and 7 low expression in cases of gastric cancer. The frequency of high expression cases was greater in gastric cancer than in esophageal or colorectal cancers (p < 0.05). There were no differences in clinicopathologic factors including prognosis between high and low expression cases of each cancer. Our mRNA study of heparanase indicated that its expression status was different among gastrointestinal cancers. The clinical and pathological impact was low compared to other proteinases, although further studies are recommended for final conclusion.
Asunto(s)
Adenocarcinoma/enzimología , Carcinoma de Células Escamosas/enzimología , Neoplasias Colorrectales/enzimología , Neoplasias Esofágicas/enzimología , Glucuronidasa/genética , Neoplasias Gástricas/enzimología , Adenocarcinoma/patología , Anciano , Carcinoma de Células Escamosas/patología , Neoplasias Colorrectales/patología , Cartilla de ADN/química , Neoplasias Esofágicas/patología , Femenino , Glucuronidasa/metabolismo , Humanos , Masculino , ARN Mensajero/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias Gástricas/patología , Células Tumorales CultivadasRESUMEN
Tumor angiogenesis progresses by a dynamic balance between tumor vascular regression and growth. Angiopoietin (Ang)-2 (the natural antagonist for the angiogenic Tie-2 receptor) and vascular endothelial growth factor (VEGF) are thought to be critical regulators in this process; therefore, these may play a critical role in cancer aggressiveness. The aim of this study was to clarify the clinical and biological significance of the expression of Ang-2 in human gastric cancers and to investigate the relationship between Ang-2 together with VEGF and the induction of proteases such as matrix metalloproteinases (MMPs) in the process of tumor development. Eighty-five individuals with gastric cancer, who had undergone surgery without preoperative treatment, were studied. A stable transfectant of the human MKN-7 gastric cancer cell lines with an Ang-2 expression vector was used for the experimental study. First, we examined the relationship between the mRNA expression of Angs by Northern blot analysis and clinicopathological features. High Ang-2-expression cases showed more frequent vascular involvement and more advanced stages of disease compared with low Ang-2-expression cases (P < 0.05). With regard to prognosis, the survival time for patients in the high-Ang-2 mRNA group was significantly shorter (P < 0.05). When we examined the localization of Ang-2 in human gastric cancers, immunohistochemical analysis revealed that this protein was expressed predominantly in cancer tissues when compared with normal tissues. Interestingly it was expressed not only in endothelia cells (ECs) but also in cancer cells. Second, Ang-2-transfected cells were implanted in vivo into the gastric walls of nude mice. Ang-2-transfectant mice developed highly metastatic tumors with hypervascularity as compared with MKN-7 or control vector-transfectant tumors. There was a significant correlation between Ang-2 mRNA expression and lower grade of vessel maturation. Third, on the basis of the in vivo data, we focused on production of proteases such as MMPs to investigate possible mechanisms in these processes. MMP-1, MMP-9, and urokinase-type plasminogen activator in ECs were strongly up-regulated by Ang-2 in the presence of VEGF in vitro. These data suggest that production of Ang-2 is implicated in tumor development in human gastric cancers. Its production may contribute to tumor angiogenesis by induction of proteases in ECs, which may be enhanced in the presence of VEGF.
Asunto(s)
Metaloproteinasas de la Matriz/biosíntesis , Neovascularización Patológica/enzimología , Proteínas/fisiología , Neoplasias Gástricas/irrigación sanguínea , Angiopoyetina 2 , Animales , Factores de Crecimiento Endotelial/fisiología , Endotelio Vascular/enzimología , Femenino , Humanos , Inmunohistoquímica , Linfocinas/fisiología , Masculino , Metaloproteinasas de la Matriz/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Neovascularización Patológica/metabolismo , Biosíntesis de Proteínas , Proteínas/genética , Proteínas Tirosina Quinasas Receptoras/antagonistas & inhibidores , Receptor TIE-2 , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias Gástricas/enzimología , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Transfección , Células Tumorales Cultivadas , Activador de Plasminógeno de Tipo Uroquinasa/biosíntesis , Activador de Plasminógeno de Tipo Uroquinasa/metabolismo , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial VascularRESUMEN
OBJECTIVE: N-Acetylation polymorphism is a representative genetic trait related to an individual's susceptibility to several cancers. However, there remains a controversy and no consensus concerning whether there is a true association between esophageal cancer and N-acetylation polymorphism. METHODS: To analyze the distribution of N-acetyltransferase 2 polymorphism in Japanese patients with esophageal squamous cell cancer, a molecular genotyping method using a polymerase chain reaction-based restriction fragment length polymorphism was used. RESULTS: Based on an analysis of 71 Japanese patients with esophageal squamous cell cancer and 329 healthy control subjects, the distribution of the slow acetylator phenotype was significantly higher in esophageal cancer patients than in the controls (19.7% and 9.4%, respectively, p = 0.040). The odds ratio of esophageal cancer for the slow phenotype was 2.55 (95% CI = 1.15-5.65, p = 0.023) compared with the rapid type. Furthermore, a significant difference between the distribution of acetylator phenotype and the incidence of lymph node metastasis and lymphatic involvement was found based on the clinicopathological features of these cancers. Esophageal cancer patients with a higher smoking exposure history tended to have the rapid acetylator phenotype. CONCLUSION: These results suggest that N-acetylation polymorphism may be implicated as a genetic trait affecting an individual's susceptibility and biological behavior of esophageal squamous cell cancer.
Asunto(s)
Arilamina N-Acetiltransferasa/genética , Carcinoma de Células Escamosas/genética , Neoplasias Esofágicas/genética , Polimorfismo Genético , Anciano , Alelos , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad/genética , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Valores de ReferenciaRESUMEN
Tissue inhibitor of metalloproteinase-1 (TIMP-1) inhibits the activity of matrix metalloproteinase, which may play an important role in carcinoma invasion and metastasis. TIMP-1 is thus considered to inhibit carcinoma invasion and metastasis. However, TIMP-1 possesses another important function, cell growth promotion. The clinical significance of TIMP-1 expression has not been fully determined in esophageal carcinoma. We thus examined the expression of TIMP-1 mRNA in tumor (T) and corresponding normal (N) tissues of 85 esophageal carcinoma cases by RT-PCR. The T:N ratio of TIMP-1 mRNA expression in each case was evaluated semi-quantitatively with adjustment by an internal control gene. The mean T:N ratio was 2.0 (range 0.2-6.5). When comparing high-expression cases (T:N > 2.0, n = 37) with low-expression cases (T:N < or = 2.0, n = 48), the former showed a significantly higher frequency of lymph vessel invasion, vascular vessel invasion, lymph node metastasis and advanced-stage disease. The former cases showed a poorer prognosis than the latter. Multivariate analysis disclosed that TIMP-1 expression status was an independent determining factor for prognosis. Our findings suggest that TIMP-1 expression correlates with tumor extension of esophageal carcinoma and might, if validated, prove useful as a novel prognostic marker for esophageal carcinoma.
Asunto(s)
Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patología , Inhibidor Tisular de Metaloproteinasa-1/análisis , Inhibidor Tisular de Metaloproteinasa-1/genética , Anciano , Neoplasias Esofágicas/mortalidad , Neoplasias Esofágicas/cirugía , Femenino , Estudios de Seguimiento , Humanos , Metástasis Linfática , Masculino , Invasividad Neoplásica , Estadificación de Neoplasias , Pronóstico , ARN Mensajero/análisis , Análisis de Regresión , Estudios Retrospectivos , Tasa de Supervivencia , Factores de Tiempo , Transcripción GenéticaRESUMEN
OBJECTIVES: Interleukin-8 (IL-8) as an alpha-chemokine recruits and activates neutrophils, which are abundant in the intestinal lesions of ulcerative colitis (UC) and Crohn's disease (CD). Stromal cell-derived factor 1 (SDF1-alpha) is a new chemokine that is chemotactic to neutrophils. The aims of this study were to assess the relative expression of SDF1-alpha and IL-8 mRNA in different colonic regions and patients with inflammatory bowel disease with varied degrees of inflammation in the colon. METHODS: Colon biopsy samples were obtained from 19 patients with UC, 12 with CD, and 5 with irritable bowel syndrome (IBS) who underwent colonoscopy. Levels of IL-8 and SDF1-alpha mRNA expression were measured semiquantitatively by reverse-transcription and polymerase chain reaction amplification. The cytokine mRNA levels were corrected for glyceraldelyde-3-phosphate dehydrogenase mRNA expression. RESULTS: IL-8 mRNA expression was significantly correlated with SDF1-alpha expression in normal biopsies from IBS patients (r = 0.58, p < 0.01). There was no significant difference in cytokine mRNA expression (IL-8 or SDF1-alpha) across different regions of the colon or rectum in uninflamed normal biopsies. The IL-8 mRNA expression ratios in UC (mean +/- SD, 1.03 +/- 0.52) and CD (0.90 +/- 0.38) patients were significantly higher than in IBS (0.52 +/- 0.17) (p < 0.01, p < 0.05, respectively). The SDF1-alpha mRNA expression ratio in UC (0.30 +/- 0.52) was higher than in both CD (0.21 +/- 0.10) and IBS patients (0.22 +/- 0.11) (p < 0.01, <0.05, respectively). A statistically significant correlation was found between the IL-8 mRNA expression and the colonic inflammation in UC patients (r = 0.44, p < 0.05) but not for SDF1-alpha expression in UC patients. CONCLUSIONS: IL-8 but not SDF1-alpha mRNA expression was associated with inflammation in UC. This suggests that IL-8 may play a more important role in inflammatory bowel disease than does SDF1-alpha.
Asunto(s)
Quimiocinas CXC/biosíntesis , Colitis Ulcerosa/metabolismo , Enfermedad de Crohn/metabolismo , Interleucina-8/biosíntesis , Adulto , Biopsia , Estudios de Casos y Controles , Quimiocina CXCL12 , Colon/metabolismo , Colon/patología , Enfermedades Funcionales del Colon/metabolismo , Femenino , Gliceraldehído-3-Fosfato Deshidrogenasas/biosíntesis , Humanos , Masculino , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células del EstromaRESUMEN
Tumor angiogenesis is essential for tumor growth and tumor metastasis, and it depends on angiogenic factors produced by tumor cells and/or infiltrating cells in tumor tissue. In this study, we evaluated the clinical significance of the expression of angiogenin, which is a potent angiogenic protein, and the relationship between its mRNA expression and focal macrophage infiltration in colorectal cancer. Furthermore, we investigated the induction of angiogenin mRNA expression by proinflammatory cytokines mainly produced by inflammatory cells in tumor tissues. When we examined the relationship between the mRNA expression of angiogenin, by semiquantitative reverse transcription-PCR, and clinicopathological features in 65 patients with colorectal cancer, there was a significant difference in the vascular involvement, lymph node metastasis, liver metastasis, and advanced stage in patients with high-expression of angiogenin compared with low expression (P < 0.05). With regard to prognosis, the survival time for subjects in the high angiogenin mRNA group (tumor:normal ratio >1.9) was significantly worse (P < 0.05). When we examined the localization of angiogenin in colorectal cancer, immunohistochemical analysis in 65 patients with colorectal cancer revealed that angiogenin was predominantly expressed in cancer cells compared with stromal cells or normal tissues. The intensity of staining of angiogenin was significantly correlated with microvessel counts and focal macrophage infiltration counts (P < 0.05). In an in vitro study, interleukin-1beta and tumor necrosis factor-alpha induced angiogenin mRNA expression in colon cancer cells in a dose- and time-dependent manner, and these cytokines significantly upregulated the expression of angiogenin mRNA, especially in colon cancer cells rather than in other cells in the stroma of tumor tissues (fibroblasts, tumor infiltrating lymphocytes, macrophages). These results suggest that tumor angiogenesis in colorectal cancer may be advanced, at least in part, by angiogenin induced by proinflammatory cytokines derived from infiltrating macrophages.
Asunto(s)
Neoplasias Colorrectales/metabolismo , Macrófagos/fisiología , Ribonucleasa Pancreática/biosíntesis , Neoplasias Colorrectales/irrigación sanguínea , Neoplasias Colorrectales/genética , Femenino , Expresión Génica , Células HT29/efectos de los fármacos , Células HT29/metabolismo , Humanos , Inmunohistoquímica , Interleucina-1/biosíntesis , Interleucina-1/farmacología , Mucosa Intestinal/irrigación sanguínea , Mucosa Intestinal/metabolismo , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Persona de Mediana Edad , Neovascularización Patológica/metabolismo , Neovascularización Fisiológica/efectos de los fármacos , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ribonucleasa Pancreática/genética , Factor de Necrosis Tumoral alfa/biosíntesis , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
Tissue inhibitor of metalloproteinase-1 (TIMP-1) is an inhibitor of matrix metalloproteinase (MMP) in human carcinomas. TIMP-1 is thus considered to inhibit carcinoma invasion and metastasis. On the other hand, recent reports have disclosed that TIMP-1 also possesses a growth-promoting function. The clinical significance of TIMP-1 expression has not been fully determined in breast carcinoma. We thus examined the expression of TIMP-1 mRNA in tumor tissues of 100 breast carcinoma cases by a reverse transcriptase-polymerase chain reaction assay. The expression of TIMP-1 mRNA in each case was evaluated semi-quantitatively with adjustment for the TIMP-1 expression in a control breast carcinoma cell line, MCF7. There was a significant inverse correlation between the TIMP-1 expression and lymph node metastasis (p<0.05). A multivariate analysis disclosed that TIMP-1 expression status was an independent determinant factor for lymph node metastasis. In addition, the tumors with positive estrogen or progesterone receptors showed a higher TIMP-1 mRNA expression than those without the receptors, but this did not reach statistical significance. The findings suggest that the breast tumors with high TIMP-1 expression might show less malignant potential than those with low TIMP-1 expression.
Asunto(s)
Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Inhibidor Tisular de Metaloproteinasa-1/análisis , Inhibidor Tisular de Metaloproteinasa-1/genética , Adulto , Factores de Edad , Anciano , Neoplasias de la Mama/cirugía , Femenino , Estudios de Seguimiento , Humanos , Metástasis Linfática , Persona de Mediana Edad , Invasividad Neoplásica , Estadificación de Neoplasias , Valor Predictivo de las Pruebas , ARN Mensajero/análisis , Receptores de Estrógenos/análisis , Receptores de Progesterona/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Transcripción Genética , Células Tumorales CultivadasRESUMEN
BACKGROUND: Collagenase-3 (matrix metalloproteinase-13, MMP-13) is a recently identified human MMP with broad substrate specificity which can be activated by membrane type 1 (MT1) matrix metalloproteinase in vitro. These may play a critical role in cancer aggressiveness. AIMS: To examine the clinical significance of collagenase-3 expression and the cooperative role of MT1-MMP in human oesophageal carcinomas. PATIENTS: Forty five individuals with oesophageal carcinoma who underwent surgery without preoperative treatment. METHODS: The tumour/normal (T/N) ratios of collagenase-3 and MT1-MMP mRNA expression in 45 human oesophageal carcinomas were determined by northern blot analysis. The production and localisation of collagenase-3 and MT1-MMP proteins were investigated by immunohistochemistry, western blot analysis, and zymography. RESULTS: The mean T/N ratio of collagenase-3 mRNA was 3.5 and that of MT1-MMP 2.1. There was a significant correlation between collagenase-3 and MT1-MMP mRNA expression (p<0.001). Twenty two cases with a collagenase-3 T/N ratio >3.5 showed a significantly higher frequency of vascular involvement and lymph node metastasis, and tended to be at a more advanced stage than 23 cases with a T/N ratio < or =3.5 (p<0.05). Western blot analysis and zymography demonstrated production of collagenase-3 protein in tumour tissues but not in normal tissues. Immunohistochemical studies revealed that collagenase-3 was localised predominantly in tumour cells and MT1-MMP was detected in the same collagenase-3 positive cells; there was a significant association between collagenase-3 and MT1-MMP protein expression (p<0.05). With regard to prognosis, the survival time for subjects in the high collagenase-3 group (T/N ratio >3.5) was significantly worse (p<0.05). CONCLUSIONS: These data suggest that production of collagenase-3 together with MT1-MMP is implicated in tumour aggressiveness and prognosis in human oesophageal carcinomas.
Asunto(s)
Colagenasas/metabolismo , Neoplasias Esofágicas/enzimología , Metaloendopeptidasas/metabolismo , Proteínas de Neoplasias/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Northern Blotting , Western Blotting , Colagenasas/genética , Neoplasias Esofágicas/patología , Neoplasias Esofágicas/cirugía , Femenino , Estudios de Seguimiento , Expresión Génica , Humanos , Técnicas para Inmunoenzimas , Masculino , Metaloproteinasa 13 de la Matriz , Metaloproteinasas de la Matriz Asociadas a la Membrana , Metaloendopeptidasas/genética , Persona de Mediana Edad , Pronóstico , ARN Mensajero/genética , ARN Neoplásico/genética , Tasa de Supervivencia , Resultado del TratamientoRESUMEN
Sentinel lymph node (SLN) biopsy is being evaluated in breast cancer patients to improve detection of metastases and to guide therapy with minimal morbidity. The aim of this study was to increase the sensitivity of tumor cell detection in SLNs using superior reverse transcription polymerase chain reaction (RT-PCR) for carcinoembryonic antigen (CEA) and mammaglobin (MMG) analysis rather than current methods which fail to identify clinically relevant disease in many patients. In seventy stage I and II breast cancer patients dye-guided lymph node mapping was performed and the SLNs were divided alternately for RT-PCR or hematoxylin and eosin staining (H&E). RT-PCR and H&E diagnosis of SLNs were compared. SLNs were identified in 66/70 (94.3%) patients. Seventeen patients (26. 2%) had histological metastasis in SLNs. CEA was expressed in 25.0% of 48 patients with H&E negative SLNs, and MMG was expressed in 20. 8%. SLNs could predict axillary lymph node status with 95.4% accuracy and 6.3% false negative rate by H&E. Moreover, RT-PCR improved these to 98.5% and 2.8%, respectively. SLN diagnosis using RT-PCR is a powerful and sensitive method, which increases the accuracy of clinical staging and may provide more informed choices for appropriate therapeutic management of breast cancer patients.
Asunto(s)
Neoplasias de la Mama/metabolismo , Antígeno Carcinoembrionario/metabolismo , Proteínas de Neoplasias/metabolismo , Uteroglobina/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias de la Mama/patología , Femenino , Humanos , Metástasis Linfática , Mamoglobina A , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sensibilidad y EspecificidadRESUMEN
L-myc polymorphism is a representative genetic trait related to an individual's susceptibility to several cancers. However, there have been no reports concerning the association between esophageal cancer and L-myc polymorphism. To analyze the distribution of polymorphism in Japanese patients with esophageal cancer, a molecular genotyping method using a polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) was used. Based on an analysis of 65 Japanese patients with esophageal cancer and 107 healthy control subjects, a significant difference was observed in either the distribution of genotypes (P=0.012) or of allele frequencies between the two groups (P=0.004). The relative risk of esophageal cancer for genotypes including the shorter allele was 2.9 compared to the longer allele homozygote. Furthermore, the patients with S-allele had a tendency for poor prognosis among those with three genotypes. A significant difference between the distribution of genotypes and the incidence of lymph node metastasis was found based on the clinicopathological features of the cancers. These results suggest that L-myc polymorphism may be implicated as a genetic trait affecting an individual's susceptibility to esophageal cancer, at least among Japanese patients.
Asunto(s)
Neoplasias Esofágicas/genética , Genes myc , Polimorfismo de Longitud del Fragmento de Restricción , Neoplasias Esofágicas/mortalidad , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Tasa de SupervivenciaRESUMEN
One of the genes that the authors have isolated by a differential display of hepatocellular carcinoma compared to adjacent liver is the alpha6 integrin. alpha6 integrin is the major adhesion receptor for laminin and is suggested to be involved in tumor cell invasion and metastasis. To our knowledge, however, there are no reports on alpha6 integrin expression in esophageal carcinoma. We thus conducted a study to determine its clinicopathologic significance in human esophageal carcinoma. The tumor/normal (T/N) ratio of alpha6 integrin expression was calculated by Northern hybridization in 45 cases of esophageal carcinoma. In selected cases the expression of the alpha6 integrin variants A and B was also investigated by reverse transcriptase-polymerase chain reaction, and immunostaining for alpha6 integrin was performed. The expression levels of alpha6 integrin mRNA in the tumor tissue were greater than those of the corresponding normal tissue in 39 of 45 cases (87%). The overexpression of alpha6 integrin was also recognized by immunostaining. Fifteen cases with a high T/N ratio demonstrated a deeper invasion into the esophageal wall than the 30 cases with a low T/N ratio. Although there was no significant difference, the 15 cases with a high T/N ratio had a tendency for a worse prognosis. The ratio of the two variants (alpha6A/alpha6B) did not show any relationship to survival. The findings imply that alpha6 integrin is overexpressed in human esophageal carcinomas and that alpha6 integrin may play an important role in esophageal tumor invasion.
Asunto(s)
Antígenos CD/análisis , Neoplasias Esofágicas/química , Adulto , Anciano , Northern Blotting , Neoplasias Esofágicas/mortalidad , Neoplasias Esofágicas/patología , Femenino , Humanos , Inmunohistoquímica , Integrina alfa6 , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Our aim was to assess whether, in the United States, with the predominant hepatitis C viral (HCV) genotypes 1a/I and 1b/II, hepatic interferon-alpha receptor (IFNAR) mRNA expression correlated with response to IFN therapy, levels of HCV RNA, or histologic activity index (HAI). Nine of 24 patients (38%) had an initial response to IFN treatment, 5 of whom (21%) had a sustained response. The corrected hepatic IFNAR mRNA expression (measured by RT-PCR) for the sustained responder group (mean +/- SE, 0.16 +/- 0.06, n = 5) was significantly higher than for the nonresponding group (0.059 +/- 0.01, n = 15) (p < 0.02). Patients who relapsed had an intermediate value (0.092 +/- 0.029, n = 4). Higher IFNAR expression was inversely correlated with a lower serum HCV RNA titer (p < 0.01), and responders to IFN treatment tended to have a lower titer of HCV RNA (p = 0.056). We found no significant correlation between the amounts of IFNAR with (1) the total HAI (low HAI < or = 7, IFNAR 0.076 +/- 0.013, n = 10; high HAI > or = 8, IFNAR 0.092 +/- 0.027, n = 14, ns) or (2) individual inflammation, necrosis, or fibrosis components of the HAI. As with Japanese HCV patients with genotypes 1b/II-2b/IV, higher hepatic IFNAR mRNA expression in the United States with predominant genotypes 1a/I and 1b/II appears to correlate with response to IFN therapy and a low HCV RNA titer but not with the total HAI or its components.
Asunto(s)
Antivirales/uso terapéutico , Hepacivirus/genética , Hepatitis C Crónica/patología , Interferón-alfa/uso terapéutico , ARN Mensajero/biosíntesis , Receptores de Interferón/genética , Adulto , Biopsia , Femenino , Genotipo , Humanos , Interferón alfa-2 , Hígado/patología , Masculino , Receptor de Interferón alfa y beta , Proteínas Recombinantes , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Resultado del Tratamiento , Estados UnidosRESUMEN
We have recently demonstrated by Northern blot and RT-PCR that the mRNA expression of the alpha-chemokine hIRH/SDF-1alpha is reduced in hepatocellular carcinoma (HCC), several digestive tract cancers and premalignant colon adenomas, and that its receptor CXCR4 mRNA expression is reduced in HCC. Here we investigate the expression of CXCR4 mRNA expression in several digestive tract cancers and hepatitis C viral (HCV) infected liver, a premalignant condition. There was no difference in the CXCR4 mRNA expression in colon, esophageal or gastric cancers compared to non-cancerous tissues. This is significantly different from the reduced expression we have seen with hepatocellular carcinoma (p<0.05). To better refine regional tumor or hepatic cytokine mRNA analysis within a biopsy sample we describe a micro-isolation technique for RNA extraction from portal and triad areas of liver biopsies or other small malignant or non-malignant biopsy samples suitable for use in RT-PCR and differential display reactions. In HCV liver biopsies, the expression of hIRH and its receptor CXCR4 mRNA, corrected for G3PDH, was not significantly different from that of control non-HCV (steatosis) biopsies. CXCR4 is expressed on leukocytes and its expression was predicted to correlate with hepatic inflammation. CXCR4 receptor mRNA expression did correlate significantly with that of its ligand hIRH/SDF-1alpha (p=0.001), and with the severity of fibrosis (p<0.05), but not with portal inflammation (p<0.10), piecemeal necrosis (p<0.10), lobular inflammation (p>0.10), the presence of lymphoid aggregates (p>0.10), or the total histological activity index (p=0.07). There was no difference in expression of hIRH or CXCR4 between responders and non-responders to interferon (IFN) treatment, while as a control, the responder group of patients did show a higher expression of IFNalpha receptor than the non-responder group (p=0.05).
Asunto(s)
Neoplasias del Colon/metabolismo , Neoplasias Esofágicas/metabolismo , Hepatitis C/metabolismo , Hígado/metabolismo , Receptores CXCR4/biosíntesis , Neoplasias Gástricas/metabolismo , Biopsia , Quimiocina CXCL12 , Quimiocinas CXC/biosíntesis , Citocinas/biosíntesis , Hepacivirus/metabolismo , Hepatitis C/virología , Humanos , Hígado/virología , ARN Mensajero/biosíntesis , ARN Neoplásico/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosRESUMEN
Differential cDNA displays between hepatocellular carcinoma and adjacent non-malignant tissues have previously detected a PCR product, hIRH (human intercrine reduced in hepatomas), equivalent to SDF1alpha/PBSF whose mRNA was lost from human hepatocellular carcinoma and other malignant and pre-malignant samples and malignant cell lines. There are no reports to date of the mRNA status of the receptor for hIRH/SDF1alpha/PBSF, CXCR4 in malignant tissues. We report here that there is a reduction in the mRNA expression of CXCR4 in hepatocellular carcinoma as estimated by Northern blot and RT-PCR and compared to the adjacent non-malignant tissue. The average (mean SD) tumor/normal ratio for CXCR4 mRNA expression, determined by RT-PCR, was 0.65 0.36 in 10 pairs of hepatocellular carcinomas. There was no consistent loss of CXCR4 mRNA expression in a range of malignant cell lines. The 3'-non-coding region of hIRH, had typical early response gene element sequences. Despite the presence of these 3'-elements there was no induction of hIRH gene expression in human lung carcinoma A549 cells by tumor necrosis factor alpha, interleukin-2, lipopolysaccharide or phorbol myristic acetate, nor in human melanoma cell line SB-2 by uv irradiation, under conditions which induced the homologue CXC intercrine IL-8 expression. Furthermore, there was no induction of hIRH gene expression, but rather a suppression, upon serum or cytokine addition to serum-deprived fibroblast cell lines, to an in vitro mouse bone marrow preparation, and to monocytic cell line THP-1.
Asunto(s)
Carcinoma Hepatocelular/metabolismo , Quimiocinas CXC/biosíntesis , Neoplasias Hepáticas/metabolismo , Receptores CXCR4/biosíntesis , Animales , Quimiocina CXCL12 , Fibroblastos/metabolismo , Humanos , Interleucina-2/metabolismo , Interleucina-8/metabolismo , Hígado/metabolismo , Ratones , Monocitos/metabolismo , ARN Mensajero/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales Cultivadas , Factor de Necrosis Tumoral alfa/metabolismo , Rayos UltravioletaRESUMEN
Loss of sequences from human chromosome 10q has been reported in several different cancers. Recently, a second candidate tumour-suppressor gene, DMBT1, was identified in this chromosomal region. We studied the mRNA expression and homozygous deletion of this gene in human oesophageal, gastric and colon cancers. Reverse transcriptase polymerase chain reaction (RT-PCR) amplification demonstrated that 23 (53.5%) of 43 oesophageal, 5 (12.5%) of 40 gastric, and 4 (16.7%) of 24 colorectal cancer cases showed an apparent reduction in DMBT1 mRNA in tumour tissues compared with paired normal tissues. Twelve out of 15 oesophageal cancer cell lines also showed no expression. We next studied homozygous deletions within the DMBT1 gene in oesophageal cancers by using duplex PCR. Consequently, it was recognized in five (11.6%) of the primary tumours and two (13.3%) of the cell lines. These findings suggest that DMBT1 may act as a tumour-suppressor gene not only in brain tumours but also in gastrointestinal cancers, especially in oesophageal cancers.
Asunto(s)
Aglutininas , Cromosomas Humanos Par 10/genética , Neoplasias Esofágicas/genética , Neoplasias Gastrointestinales/genética , Genes Supresores de Tumor , Proteínas de Neoplasias/metabolismo , Receptores de Superficie Celular/metabolismo , Proteínas de Unión al Calcio , Neoplasias Colorrectales/genética , Proteínas de Unión al ADN , Humanos , Proteínas de Neoplasias/genética , ARN Mensajero/metabolismo , Receptores de Superficie Celular/genética , Neoplasias Gástricas/genética , Proteínas Supresoras de TumorRESUMEN
Angiogenic factors such as vascular endothelial growth factor (VEGF) may be involved in neovascularization of malignant tumors. Our aim was to determine whether there is an increased VEGF mRNA expression in liver from patients with HCC and premalignant hepatitis C virus (HCV) with differing severity of inflammation. VEGF mRNA (VEGF165, VEGF189) was detected by reverse transcription and semi-quantitative polymerase chain reaction (RT-PCR) amplification in all liver samples. There was no difference in VEGF mRNA expression ratios (corrected for glyceraldehyde-3-phosphate dehydrogenase) among three groups: steatohepatitis, as a non-malignant non-viral control, 1. 05+/-0.35, n=8; chronic hepatitis C, 0.86+/-0.27, n=18; hepatocellular carcinoma, 1.06+/-0.43, n=10. VEGF mRNA expression was independent of the severity of HCV inflammation estimated by the histological activity index: low HAI (=4, n=8) vs. high HAI (>/=10, n=10), 0.93+/-0.31 vs. 0.81+/-0.24, p=ns. There was no significant difference in mean VEGF expression between HCC tumor (1.06+/- 0.43) and adjacent tissue (0. 85+/-0.42) although the tumors tended to have higher expression than adjacent non-malignant tissues. In conclusion, all liver samples of steatohepatitis, chronic HCV infection and HCC expressed VEGF mRNA, VEGF mRNA may be uniformly expressed in liver tissue, the level of expression is probably not related to virus infection or the severity of inflammation. Other angiogenic or angiostatic factors might be more involved in angiogenesis in HCC.