metallothionein and Thymoma

Metallothioneins (MTs) are low molecular weight and cysteine-rich intracellular proteins involved in metal homeostasis and detoxication. They are found in certain normal tissues, and are overexpressed in various tumours with correlation to more aggressive behaviour in certain tumours. Since the histopathological types of thymoma have unpredictable invasive potential, MT over-expression was investigated as a possible marker of the invasive potential of thymomas. We studied immunohistochemical MT expression in 27 non-invasive thymomas, 20 micro-invasive thymomas, and 23 macro-invasive thymomas with a mouse monoclonal anti-MT antibody E9 on formalin-fixed paraffin-embedded tissues. MT expression was significantly different among the three groups of thymomas (P = 0.02) with a stronger expression in invasive thymomas (P = 0.003). However, MT expression was not exclusively limited to invasive thymomas. Therefore, it could not be used as a marker of aggressive potential in individual thymomas. Analysis of MT expression according to the histological types of the thymomas revealed that eight of nine spindle cell thymomas, none of 10 small polygonal cell thymomas, four of 14 mixed thymomas, seven of 29 large polygonal cell thymomas, and seven of eight squamoid thymomas significantly expressed MT. There was a statistically significant difference in MT expression among different histological types of thymomas (P = 0.000). The strongest and most consistent expression was observed in spindle cell thymoma and squamoid thymoma. Since spindle cell thymoma was usually non-invasive and squamoid thymoma was more aggressive, MT expression does not correlate with the invasive potential of different histological types of thymomas. But because medullary epithelial cells of the thymus were positive for MT, our results suggest that both spindle cell thymoma and squamoid thymoma might derive from the medullary compartment of the thymus.

Topics: Animals; Biomarkers, Tumor; Humans; Immunohistochemistry; Metallothionein; Mice; Thymoma

Metallothionein functions as a radical scavenger protecting cells from the indirect effect of radiations. We investigated the effect of bismuth nitrate, an efficient inducer of metallothionein, on acute and late effects of radiation in mice. Metallothionein contents were examined in several organs after the administration of bismuth nitrate. The content in bone marrow increased 2-fold in the treated as compared to the control mice. This treatment protected irradiated mice from bone marrow death and increased the number of endogenous spleen colonies. The metallothionein content in the ileum did not change after treatment with bismuth nitrate. Mice were not protected by bismuth nitrate when exposed to 9 Gy of X-rays. This suggests that this agent does not protect from gastrointestinal death. The incidence of X-ray-induced thymic lymphomas was lowered by the administration of bismuth nitrate in mice exposed to four fractionated doses of 1.3 Gy of X-rays. These results indicate that bismuth nitrate effectively modified both acute and late effects of X-rays by inducing metallothionein in the target tissues.

Topics: Animals; Bismuth; Metallothionein; Mice; Mice, Inbred C57BL; Neoplasms, Radiation-Induced; Nitrates; Radiation-Protective Agents; Thymoma; Thymus Neoplasms

Using an immunofluorescence (IF) assay, the presence of metallothionein (MT) was investigated in sections of normal and pathologic human thymuses as well as in cultures of thymic epithelial cells. This protein, known to have a high binding affinity for class II B transitional metals, such as zinc, was detected in the epithelial component of the thymus. Moreover, double labeling experiments with the anti-MT and an anti-thymulin monoclonal antibody showed that all cells containing thymulin, a thymic hormone whose active structure is known to contain zinc, also exhibited large amounts of metallothionein. These results, together with the fact that zinc and thymulin have been detected in the same type of cell organelles, lead to the conclusion that the MT present in thymic epithelial cells might be involved in the mechanism of zinc storage in these cells, thus favoring the secretion of thymulin in its biologically active, zinc-containing form.

Topics: Animals; Biological Assay; Child, Preschool; Cross Reactions; Fluorescent Antibody Technique; Humans; Immune Sera; Infant; Metallothionein; Mice; Mice, Inbred C57BL; Thymic Factor, Circulating; Thymoma; Thymus Gland; Thymus Hormones; Thymus Neoplasms; Zinc

The W7 mouse thymoma cell line does not express the metallothionein-I (MT-I) gene in the presence of either cadmium or glucocorticoids, unlike most other cell lines. This cell line was therefore used as a model system for studying the role of DNA methylation on MT-I gene expression. The extent of DNA methylation within the MT-I gene and its flanking regions was determined by comparing the cleavage patterns generated by the isoschizomeric restriction enzymes Hpa II and Msp I. In W7 cells, all of the Hpa II sites in the vicinity of the MT-I gene are methylated, whereas in cells that have an expressible MT-I gene (for example, Friend erythroleukemia cells) all of these Hpa II sites are unmethylated. When W7 cells are treated for a few hours with 5-azacytidine, the MT-I gene becomes inducible by both cadmium and glucocorticoids. Addition of hydroxyurea along with 5-azacytidine prevents MT-I gene induction, suggesting that incorporation of 5-azacytidine into DNA is required before this gene can be activated. To determine whether 5-azacytidine treatment changes the methylation pattern near the MT-I gene, we treated W7 cells with 5-azacytidine and selected inducible cells in 10 micro M cadmium. all of the Hpa II sites within the MT-I gene are unmethylated in these cadmium-resistant W7 cells. In addition, flanking DNA sequences are also undermethylated in a pattern similar to that seen in Friend erythroleukemia cells that express the MT-I gene. The possible significance of methylation as a mechanism of gene commitment during cell differentiation is discussed.

Topics: Animals; Azacitidine; Cadmium; Cell Line; Dexamethasone; DNA; Gene Expression Regulation; Metalloproteins; Metallothionein; Methylation; Mice; RNA, Messenger; Thymoma; Transcriptional Activation