RESUMEN
Substantial progress has been make in elucidating the biochemical properties of HSPs (Heat Shock Proteins) as molecular chaperones in protein biogenesis and their roles in thermoprotection and cytoprotection against cellular insults. Recently, besides advantageous cellular functions, the detrimental role of HSPs has been implicated in a variety of diseases. HSP47 is assumed to be a collagen specific molecular chaperone and its involvement in the progression of fibrosis was observed. There have been a number of reports that suggest the role of HSP60 as an autoantigen in a variety of autoimmune diseases. PSK, a protein-bound polysaccharide, is a biological response modifier that is clinically used for the treatment of cancer patients in Japan. PSK shows a broad range of biological effects in addition to the antitumor activity. In this study, we evaluated the effect of PSK on the expression of HSPs in human tumor strains at the protein and mRNA levels. PSK was observed to suppress the expression of HSP47 and HSP60 but not HSP72/73 in human tumor cell lines. Thus, the novel pharmacological potential of PSK was suggested in the diseases derived from the aberrant expression of HSPs.
Asunto(s)
Antibióticos Antineoplásicos/farmacología , Chaperonina 60/genética , Proteínas de Choque Térmico/genética , Proteoglicanos/farmacología , Animales , Northern Blotting , Western Blotting , Neoplasias de la Mama , Chaperonina 60/análisis , Neoplasias del Colon , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Proteínas del Choque Térmico HSP47 , Células HeLa , Proteínas de Choque Térmico/análisis , Humanos , Neoplasias Renales , Neoplasias Pulmonares , Masculino , Neuroblastoma , Neoplasias de la Próstata , ARN Mensajero/análisis , Ratas , Ratas Endogámicas , Neoplasias Gástricas , Células Tumorales Cultivadas/química , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/metabolismoRESUMEN
While the protective role HSPs (Heat Shock Proteins) has been recognized against physiological stress such as heat shock, heavy metals and glucose starvation, recent progress has revealed another aspect of HSPs in various diseases. HSP27 has been shown to be involved in the acquired resistance of tumor cells hyperthermic and chemotherapeutic treatment. In human breast tumors, overexpression of HSP27 is associated with a shorter disease-free survival period. HSP47 is thought to be a collagen specific molecular chaperone. The involvement of HSP47 in the progression of fibrosis has been reported. Aberrant expression of HSP could cause various autoimmune diseases. Manipulation of HSP expression, therefore, could be a therapeutic target to reduce HSP-derived detrimental cellular effects. Flavonoids are a widely distributed group of plant substances, universally present in vascular plants. Although the flavonoids have been known as natural plant products as long as the alkaloids, their pharmacological effects and potential medicinal uses have been little studied by comparison. Today, the picture has changed and the biological and pharmacological activities of plant flavonoids look promising. We investigated the effect of flavonoids on the expression of HSPs in human tumor cell lines. Flavonoids inhibited the expression of HSP27, HSP47, HSP60 and HSP72/73. The results suggested the pharmacological possibilities of flavonoids in diseases derived from abnormal expression of HSPs.
Asunto(s)
Flavonoides/farmacología , Proteínas de Choque Térmico/genética , Animales , Western Blotting , Neoplasias de la Mama , Chaperonina 60/análisis , Chaperonina 60/genética , Neoplasias del Colon , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Proteínas del Choque Térmico HSP72 , Células HeLa , Proteínas de Choque Térmico/análisis , Humanos , Neoplasias Renales , Neoplasias Pulmonares , Masculino , Neuroblastoma , Neoplasias de la Próstata , ARN Mensajero/análisis , Ratas , Ratas Endogámicas , Neoplasias Gástricas , Células Tumorales Cultivadas/química , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/metabolismoRESUMEN
The cells of all organisms respond to environmental stress by synthesizing a specific group of proteins called heat shock proteins (HSPs). While the protective role of HSPs has been observed against cellular stress, recent reports have suggested a number of functions of HSPs in various cellular processes in normal and pathological conditions. HSP27 is an important low molecular weight HSP (Mr: 27,000) found in human cells. HSP27 appears to be involved in intracellular signaling and drug resistance in addition to thermotolerance. In human breast tumors, a striking association between HSP27 overexpression and a shorter disease-free survival period was reported. In this paper, we evaluated the expression levels of HSP27 in the cell lines of human breast and other tumors. Western blotting of HSP27 has shown the correlation of HSP27 expression and metastasis and tumorigenicity of mammary and prostate tumors. Thus the biological significance of HSP27 in aggressive mammary and prostate tumors was indicated. HSP27 is suggested to be a prognostic marker for the malignancy of mammary and prostate tumors.
Asunto(s)
Proteínas de Choque Térmico/biosíntesis , Animales , Western Blotting , Neoplasias de la Mama , Neoplasias del Colon , Glioblastoma , Células HL-60/química , Células HL-60/metabolismo , Proteínas del Choque Térmico HSP72 , Células HeLa , Proteínas de Choque Térmico/análisis , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva , Neoplasias Pulmonares , Masculino , Ratones , Ratones Desnudos , Neuroblastoma , Leucemia-Linfoma Linfoblástico de Células Precursoras , Neoplasias de la PróstataRESUMEN
HSP47 is a stress-inducible glycoprotein of 47 kd molecular weight, and is assumed to be a collagen specific molecular chaperone. The expression of HSP47 is closely related to that of collagen, which is the main component of the extracellular matrix. We investigated the expression level of HSP47 in human tumor cell lines to elucidate collagen biosynthesis in tumor cells. Western blot analysis showed higher levels of HSP47 in solid tumor cell lines than in leukemia cell lines. Tumor cell lines which were derived from metastatic carcinomas and were still metastatic in animals, synthesized higher levels of HSP47. These results suggested that HSP47 may play an important role in tumor metastasis and can be a prognostic marker for the metastatic activity of human tumor cells.
Asunto(s)
Proteínas de Choque Térmico/análisis , Proteínas de Choque Térmico/biosíntesis , Animales , Western Blotting , Neoplasias de la Mama , Colágeno/biosíntesis , Colágeno/metabolismo , Neoplasias del Colon , Femenino , Glioblastoma , Células HL-60/química , Células HL-60/metabolismo , Proteínas del Choque Térmico HSP47 , Proteínas del Choque Térmico HSP72 , Células HeLa/química , Células HeLa/metabolismo , Proteínas de Choque Térmico/metabolismo , Humanos , Neoplasias Renales , Leucemia Mielógena Crónica BCR-ABL Positiva , Neoplasias Pulmonares , Masculino , Neuroblastoma , Leucemia-Linfoma Linfoblástico de Células Precursoras , Neoplasias de la Próstata , Ratas , Ratas Endogámicas , Neoplasias GástricasRESUMEN
HSP47, a heat shock-inducible glycoprotein of M(r) = 47,000, is assumed to be a molecular chaperone specific for collagen. In addition to its stress inducibility, HSP47 has transformation-sensitivity; that is, the synthesis of HSP47 decreases after malignant transformation. The expression of several HSPs in mouse fibrosarcoma, QRsP-1 and QRsP-4 were examined. HSP47 expression was weaker in the highly malignant QRsP-4 than in the weakly malignant QRsP-1, whilst HSC70 expression was similar in both cell lines. These findings suggested an important role of HSP47 as a marker of tumour malignancy of QRsP cell lines.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Fibrosarcoma/metabolismo , Proteínas de Choque Térmico/biosíntesis , Animales , Northern Blotting , Western Blotting , Fibrosarcoma/química , Proteínas del Choque Térmico HSP47 , Ratones , Trasplante de Neoplasias , ARN Neoplásico/química , Reproducibilidad de los Resultados , Células Tumorales CultivadasRESUMEN
HSP47, a collagen-binding stress protein, has transformation-sensitivity; that is, the synthesis of HSP47 decreases after malignant transformation. We examined the expression of HSP47 in transplanted tumours by Western blotting. While HSP47 was not detected in ascites tumours including Ehrlich, P388 and Sarcoma 180, marked expression of HSP47 was observed in solid tumours such as B16, B16-BL6 and Sarcoma 180. The expression of HSP47 in Sarcoma 180 cells disappeared completely after these solid tumour cells were dispersed and inoculated intraperitoneally. These findings suggested an important role of HSP47 as a marker for tumour malignancy.
Asunto(s)
Ascitis/metabolismo , Biomarcadores de Tumor/análisis , Transformación Celular Neoplásica/metabolismo , Proteínas de Choque Térmico/antagonistas & inhibidores , Sarcoma 180/metabolismo , Animales , Western Blotting , Proteínas del Choque Térmico HSP47 , Proteínas de Choque Térmico/análisis , Inyecciones Intraperitoneales , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Trasplante de NeoplasiasRESUMEN
HSP47, a heat shock-inducible glycoprotein of Mr = 47,000, is assumed to be a molecular chaperone specific for collagen. In addition to its stress inducibility, HSP47 has transformation-sensitivity; that is, the synthesis of HSP47 decreases after malignant transformation. We developed a rat monoclonal antibody against mouse HSP47, which we then used to examine the amounts of HSP47 in transplanted tumors. HSC70 (HSP73) was expressed constitutively in these ascites as well as solid tumors. While HSP47 was not expressed in ascitic form of Sarcoma 180, its expression was markedly induced during solid tumor formation of ascitic Sarcoma 180 cells after subcutaneous transplantation. The average survival period of ascites Sarcoma 180-bearing mice was 20.3 +/- 3.6 days (mean +/- SD), and was much shorter than that of solid Sarcoma 180-bearing mice. These findings suggest an important role of HSP47 in solid tumor formation as well as a possible marker for tumor malignancy.
Asunto(s)
Colágeno/metabolismo , Proteínas de Choque Térmico/biosíntesis , Sarcoma 180/metabolismo , Animales , Western Blotting , Proteínas del Choque Térmico HSP47 , Ratones , Ratones Endogámicos ICR , Trasplante de Neoplasias , Ratas , SobrevidaRESUMEN
The tumors produced by transplantation into nude mice of human adenoid squamous carcinoma-forming cell line TYS, presumably derived from a minor salivary gland, were treated with a differentiation-inducing agent, vesnarinone, which was given per o.s. daily at a dose of 200 mg/kg for 35 days. They were then examined morphologically and immunohistochemically. The vesnarinone treatment resulted in a significant suppression of tumor growth. In addition, tumor nests indicating keratinocyte and acinar cell differentiation were often observed in the treated tumors, but not in untreated controls. Tissue sections from vesnarinone-treated and untreated TYS tumors were stained with monoclonal antibody (NAb) directed to carbohydrate antigen LeY or proliferating cell nuclear antigen (PCNA) and with rabbit polyclonal antibody to p53. Antibody staining patterns were compared with morphological characteristics of cells as revealed by hematoxylin and eosin staining, and DNA fragmentation patterns as revealed by 3'-OH nick-end labelling techniques. Tissue sections from vesnarinone-treated TYS tumors showed positive reaction with nick-end labelling and were extensively stained strongly by anti-LeY MAb, whereas the untreated tumors showed negative reaction with nick-end labelling and were infrequently stained by anti-LeY MAb. Within LeY-positive areas of tissue sections from the vesnarinone-treated tumors, keratinocyte and acinar cell differentiation as well as DNA fragmentation were frequently observed, although not all LeY-positive cells showed such signs of apoptosis. LeY-positive cells showed consistent negative staining by anti-PCNA MAb and anti-p53 rabbit serum. From these findings, it can be considered that vesnarinone has differentiation and apoptosis-inducing activity against TYS cells grown in athymic nude mouse.
Asunto(s)
Antineoplásicos/uso terapéutico , Carcinoma de Células Escamosas/tratamiento farmacológico , Quinolinas/uso terapéutico , Animales , Apoptosis/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Daño del ADN , Evaluación Preclínica de Medicamentos , Humanos , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Pirazinas , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
We have developed a rat monoclonal antibody against mouse HSP47 (designated as NK47) and investigated the expression of HSP47 in several mouse ascites and solid tumors. The expression of HSP47 was not detected in ascites tumors such as P388 and Ehrlich ascites tumors while HSP47 was markedly expressed in solid tumors such as Colon-26, B16 and B16-BL6 melanomas. HSP70, HSC70 and HSP90 were expressed in almost the same level among all the tumors examined. These results suggest that the expression of HSP47 decreases as the tumor becomes more malignant.
Asunto(s)
Biomarcadores de Tumor/análisis , Proteínas de Choque Térmico/análisis , Neoplasias Experimentales/química , Animales , Carcinoma de Ehrlich , Línea Celular Transformada , Femenino , Proteínas del Choque Térmico HSP47 , Neoplasias Pulmonares/secundario , Melanoma Experimental/secundario , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Especificidad de Órganos , Ratas , Ratas Endogámicas , Especificidad de la EspecieRESUMEN
The serum and urinary ferritin levels in 52 RA patients were measured by the 2-site immunoradiometric assay method. Serum ferritin levels in RA patients correlated with C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) but not with serum iron levels and hemoglobin concentrations, although they were within the normal range. High serum ferritin levels were associated with sera with hyper gamma-globulin and rheumatoid factors. In sequential studies, serum ferritin changed in parallel with ESR, CRP and disease activity in a majority of the patients. The urinary ferritin levels and u/s ratios in some RA patients were higher than control values. Higher values were found particularly in the group of patients under gold therapy but not in groups under other treatments.