Analysis of Clonality in Endocrine Neoplasms

内分泌肿瘤的克隆性分析

• 批准号: 06671197
• 负责人: IMAI Tsuneo
• 金额: $ 1.34万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671197/
• 关键词: Clonality; MAOA; heterozygosity; Endocrine neoplasia; microsatelite; monoclonal; polyclonal; PCR

Analysis of Clonality in Endocrine Neoplasias-MAOA microsatelite methods using fluorescence primerDNA was extracted from peripheral white blood cells in female patients of various kinds of diseases. Monoamine oxidase type A (MAOA) gene was amplified by polymerase chain reaction (PCR) using fluorescence labeled primers. The amplified products were denatured and electrophoresed in 6% polyacrylamide gel and the bands were detected by automatic sequencer and analyzed using GeneScan 672 software (Applied Biosystems). The examined patients were ; aldosterone producing adenoma : 14, Cushing's syndrome (adrenal adenoma) : 5, pheochromocytoma : 1, virilizing adrenal tumor : 1, thyroid neoplasms : 4. All 24 patients' DNA were subjected to the clonal analysis. The informed cases (heterozygous for MAOA gene) were 17 of 24, so that the heterozygosity was 67%.Heterotransplantation of Human Parathyroid Glands into Nude MiceGrafts prepared from hyperplasia, adenoma, and normal glands which were resected at operation were implanted in the gluteus muscle of the recipient mice. Graft function was evaluated by measuring human intact PTH concentrations in sera of the mice. Transplantation of parathyroid tissues resulted in an increase in PTH concentrations for 4 weeks, reaching a plateau thereafter. The level remained unchanged for 8 weeks. Serum PTH concentrations were directly correlated with the volume of implanted tissues. The response of grafted parathyroid tissue to a low calcium level in sera was also observed.

采用荧光引物的MAOA微卫星法分析内分泌肿瘤的克隆性从各种疾病的女性患者外周血白细胞中提取DNA。使用荧光标记引物通过聚合酶链式反应 (PCR) 扩增 A 型单胺氧化酶 (MAOA) 基因。将扩增产物变性并在6%聚丙烯酰胺凝胶中电泳，通过自动测序仪检测条带并使用GeneScan 672软件（Applied Biosystems）进行分析。接受检查的患者是；产生醛固酮的腺瘤：14例，库欣综合征（肾上腺腺瘤）：5例，嗜铬细胞瘤：1例，男性化肾上腺肿瘤：1例，甲状腺肿瘤：4例。对所有24名患者的DNA进行了克隆分析。 24例中，通报病例（MAOA基因杂合）为17例，杂合率为67%。 人甲状旁腺裸鼠异种移植 将术中切除的增生、腺瘤和正常腺体制成移植物，植入受体小鼠的臀肌中。通过测量小鼠血清中的人类完整 PTH 浓度来评估移植功能。甲状旁腺组织移植导致 PTH 浓度增加 4 周，此后达到稳定水平。该水平连续 8 周保持不变。血清 PTH 浓度与植入组织的体积直接相关。还观察了移植的甲状旁腺组织对血清中低钙水平的反应。

项目成果

Syoichi Maruyama, Toshihide Iwashita, Tsuneo Imai, Hiroomi Funahashi, Isabella Ceccherine, Yin Luo, Giovanni Romeo, Seiichi Matsuo, Mutsushi Matsuyama, Masahide Takahashi.: "Germ Line Mutations of the ret Proto-oncogene in Japanese Patients with Multiple

Syoichi Maruyama、Toshihide Iwashita、Tsuneo Imai、Hiroomi Funahashi、Isabella Ceccherine、Yin Luo、Giovanni Romeo、Seiichi Matsuo、Matsushi Matsuyama、Masahide Takahashi。：“日本多发性硬化症患者 ret 原癌基因的种系突变

Yuji Tanaka, Hiroomi Funahashi, Tsuneo Imai, Junichi Tobinaga, Hiroshi Murase, Hiroyuki Andoh, Masaki Wada, Takako Matsuyama, Yoshihiro Taminaga, Hiroshi Takagi.: "Heterotransplantation of Human Parathyroid Glands into Nude Mice." Endocrine Journal. 42. 9

Yuji Tanaka、Hiroomi Funahashi、Tsuneo Imai、Junichi Tobinaga、Hiroshi Murase、Hiroyuki Andoh、Masaki Wada、Takako Matsuyama、Yoshihiro Taminaga、Hiroshi Takagi.：“将人甲状旁腺异种移植到裸鼠中。”

Junichi Tobinaga: "Incidentally discovered adrenocortical adenomas are not fully nonfunctioning:Immunohistochemical and dispersed adrenocortical cell study." Endocrine Journal. (in press). (1996)

Junichi Tobinaga：“偶然发现的肾上腺皮质腺瘤并非完全无功能：免疫组织化学和分散的肾上腺皮质细胞研究。”

Masaki Wada: "Detection of RET homodimers in MEN 2A-associated pheochromocytomas" Biochem Biophys Res Com. (in press). (1996)

Masaki Wada：“MEN 2A 相关嗜铬细胞瘤中 RET 同二聚体的检测”Biochem Biophys Res Com。

今井常夫: "内分泌腺腫瘍におけるClonalityの検討-MAOAマイクロサテライトにおける蛍光プライマーを用いた解析-" 名古屋大学環境医学研究所年報. 46. 123-126 (1995)

Tsuneo Imai：“内分泌肿瘤克隆性的检查 - 使用 MAOA 微卫星中的荧光引物进行分析”名古屋大学环境医学研究所年度报告 46. 123-126 (1995)。