甲状腺髄様癌および乳頭癌の培養細胞によるカルシトニン産生能の比較

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Although many groups have previously described medullary carcinoma of the thyroid gland (MCT) mainly from either clinicopathological or endocrinological viewpoints, those reports by means of an in vitro approach on the calcitonin (CT) secretion are extremely rare. We recently experienced a case with MCT (sporadic type) which underwent total thyroidectomy and bilateral modified neck dissection. Histological examination, however, showed atypical findings without stromal amyloid. Therefore, the purpose of this experiment was to confirm the final diagnosis of MCT, employing a cell-culture method for the detection of CT secretion in vitro. A small portion (0.6g of wet weight of tissue) of cancerous and non-cancerous (normal parts ascertained microscopically) tissue, removed surgically from patients with MCT or papillary carcinoma of the thyroid (PCT), were prepared for the dispersed cell culture, respectively. After the tissues were minced, they were treated with 0.05% trypsin for making dispersed cells. Incubation was carried out at 37°C in an electric incubator using Eagle's medium with 10% fetal calf serum and antibiotic solution. Thereafter, culture media were totally replaced every 4 to 6 days. Each of them was stored at - 20°C until used. The concentrations of CT in sera and culture media were measured by the use of radioimmunoassay techniques. In this study, the culture media which were replaced firstly and secondly, i.e. on the 7th and 11th day after primary culture, were utilized for CT assay. The CT levels of preoperative, postoperative 1st and 16th day's serum were 102,000, 2,400 and 200pg/ml, respectively. The serum of the 16th day was obtained 15 minutes after a calcium-infusion test, and its CT value showed less than 300pg/ml, which was the upper limit of basal serum CT in normal subjects. The concentrations of CT in the first (postcultural 7th day) and second (postcultural 11th day) culture media of medullary carcinoma cells were 299,500±29,500 (mean ± SD) and 172,000±13,000pg/ml, while those values in the papillary carcinoma were 38±21 and 25±4pg/ml, respectively. In addition, the values of CT in each of the culture media of cells from the non-cancerous tissues of MCT or PCT, were 486±78 (MCT, first) 176±60 (MCT, second), 197±2 (PCT, first) and 57±22 (PCT, second) pg/ml. The present studies demonstrated that cultured medullary carcinoma cells release substantial amounts of CT into the medium by themselves. Furthermore, there also seems to be more hyperactivity even in the non-cancerous parts of MCT compared with the normal thyroid gland. Finally, these results may prove that MCT is a calcitonin-secreting tumor.
日本内分泌学会の論文

甲状腺髄様癌および乳頭癌の培養細胞によるカルシトニン産生能の比較

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