Establishment of aspiration biopsy RNA diagnosis of malignant lymphoma

恶性淋巴瘤抽吸活检RNA诊断的建立

• 批准号: 13557227
• 负责人: TAKANO Toru
• 金额: $ 2.5万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557227/
• 关键词: mRNA; thyroid; lymphoma; PCR; monoclonality; immunoglobulin; aspiration biopsy; DNA; Thyroid; Malignant Lymphoma; ABRP; RT-PCR; IgM; Monoclonality

Distinguishing between thyroid malignant lymphoma and lymphocytic thyroiditis (Hashimoto's tbyroiditis) is 'quite difficult and problematic. Molecular techniques to detect clonal lymphoid proliferation based on immunoglobulin heavy chain (IgH) gene rearrangement may be used to facilitate more accurate diagnosis of malignant lymphoma. We tried to establish a method for diagnosing thyroid tumors by analyzing nucleic acids extracted from the needles used for fine needle aspiration biopsy. We first studied RNAs from fresh tissues samples of 8 cases of Hashimoto's thyroiditis and 18 malignant lyrnphomas to detect the monoclonality of IgH mRNA by semi-nested R.T-PCR. Monoclonality was detected in 8 of 18 (44.4%) malignant ryrnphomas but none of the 8 cases of Hashimoto's thyroiditis. We then studied aspirates from 10 cases of thyroid malignant lymphoma, 4 cases of Hashimoto's thyroiditis, and one case each of adenomatous goiter and papillary carcinoma. Monoclonality was detected in the aspirates from 4 of 10 malignant lymphomas (40%) but not from other tissues. Thus RT-PCR detection of monoclonality of IgH mRNA in addition to eytological examination may be useful in diagnosing thyroid malignant lyraphoma. In order to increase the sensitivity of this method, we examine the usefulness of degenerated oligonucleotide primed (OOP) PCR to amplify the extracted DNAs in fine needle aspiration biopsies. By mixing a primer specific for the JH sequence, DOP-PCR could produce 3-10kbp PCR product, which could be used in linker-ligation PCR or Southern blot analysis. These results may lead us to establish a low-invasive diagnostic method of malignant lymphoma in the future study by utilizing DNAs instead of RNAs obtained by fine needle aspiration biopsy

区分甲状腺恶性淋巴瘤和淋巴细胞性甲状腺炎(桥本氏结节性甲状腺炎)是非常困难和有问题的。基于免疫球蛋白重链(IgH)基因重排的克隆性淋巴增殖的分子检测技术可能有助于更准确地诊断恶性淋巴瘤。我们试图建立一种通过分析细针活检针头中提取的核酸来诊断甲状腺肿瘤的方法。用半套式RT-PCR方法检测了8例桥本甲状腺炎和18例恶性甲状腺瘤新鲜组织标本中IgH基因的单克隆性，发现8例恶性甲状腺瘤中有8例(44.4%)呈单克隆性，而8例桥本甲状腺炎中未检测到单克隆性。然后我们研究了10例甲状腺恶性淋巴瘤，4例桥本甲状腺炎，以及1例腺瘤性甲状腺肿和乳头状癌的抽吸物。10例恶性淋巴瘤中有4例(40%)抽吸物呈单克隆性，其他组织未见单克隆性。因此，除细胞学检查外，RT-PCR检测IgH基因的单克隆性可能有助于诊断甲状腺恶性淋巴管瘤。为了提高该方法的灵敏度，我们检验了简并寡核苷酸引物(OOP)聚合酶链式反应(OOP)在细针抽吸活检中扩增DNA的有效性。通过混合针对JH序列的特异引物，DOP-PCR可扩增出3-10kb的产物，可用于连接-聚合酶链式反应或Southern杂交分析。这些结果可能导致我们在未来的研究中建立一种低侵入性的恶性淋巴瘤诊断方法，利用DNA代替细针抽吸活组织检查获得的RNA

项目成果

Takano T, et al.: "Cancer-specific mRNAs in thyroid carcinomas : detection, use, and their implication in thyroid carcinogenesis"Endocr J. 49・2. 97-107 (2002)

Takano T 等人：“甲状腺癌中的癌症特异性 mRNA：检测、使用及其在甲状腺癌发生中的意义”Endocr J. 97-107 (2002)。

Hasegawa Y, Takano T, et al.: "Decreased expression of catalase mRNA in thyroid anaplastic carcinoma"Jpn J Clin Oncol. 33・1. 6-9 (2003)

长谷川Y、高野T等：“甲状腺未分化癌中过氧化氢酶mRNA的表达降低”Jpn J Clin Oncol.33·1(2003)。

Takano T et al.: "Cancer-specific mRNA in thyroid caranomas : Detection, use, and their implication in thyroid carcinogenesis"Endocrine Journal. (in press). (2002)

Takano T 等人：“甲状腺癌中癌症特异性 mRNA：检测、使用及其在甲状腺癌发生中的意义”内分泌杂志。

Takano, T, Hasegawa, Y, Miyauchi, A, Matsuzuka, F, Yoshida, H, Kuma, K, Amino, N.: "Quantitative analysis of thymosin beta-10 messenger RNA in thyroid carcinomas"Jpn J Clin Onco. 32-7. 229-232 (2002)

Takano, T, Hasekawa, Y, Miyauchi, A, Matsuzuka, F, Yoshida, H, Kuma, K, Amino, N.：“甲状腺癌中胸腺素β-10信使RNA的定量分析”Jpn J Clin Onco。

Hasegawa, Y, Takano, T, Miyauchi, A, Matsuzuka, F, Yoshida, H, Kuma, K, Amino, N.: "Decreased expression of glutathione peroxidase mRNA in thyroid anaplastic carcinoma"Cancer Lett. 182-1. 69-74 (2002)

Hasekawa, Y, Takano, T, Miyauchi, A, Matsuzuka, F, Yoshida, H, Kuma, K, Amino, N.：“甲状腺退行性癌中谷胱甘肽过氧化物酶 mRNA 的表达降低”Cancer Lett。