Establishment of tumor maker test using tumor-derived nucleic acids in plasma

利用血浆中肿瘤源性核酸建立肿瘤标志物检测

• 批准号: 14572177
• 负责人: OHTAKE Shigeki
• 金额: $ 1.86万
• 依托单位: KANAZAWA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572177/
• 关键词: Plasma DNA; PCR; Immunoglobulin gene; CDR3; NHL; 非ホジキンリンパ腫

In B cell lymphoid malignancies, malignant cells proliferate monoclonally with the same immunoglobulin gene rearrangement.This rearrangement can be detected by the use of the polymerase chain reaction(PCR).Recently, it has been reported that proliferation of clonal cells could be demonstrated by PCR analyzing DNA extracted from patients' plasma with B cell malignancies.The question, however, still remains whether amplified products were derived from tumor cells or not.In this study, using DNA extracted from both malignant cells and plasma of the same patients with B cell leukemia, I attempted to amplify immunoglobulin heavy chain(IgH) complementarity determining region 3(CDR3) by semi-nested PCR and to analyze the sequences respectively.The results showed that sequences of DNA from plasma were identical with that of DNA from tumor cells in all cases.In addition, some of them showed homology to CDR3 that was previously reported.These findings indicate that plasma DNA is released from tumor cells.In the next stage, it is necessary to raise the sensitivity, and if it works, IgH-PCR using plasma DNA might be useful tool for diagnosing patients and evaluating the efficacy of treatment, especially in patients with critical condition under which any biopsies were difficult.

在B细胞淋巴样恶性肿瘤中，恶性细胞单克隆增殖，具有相同的免疫球蛋白基因重排。这种重排可以通过使用聚合酶链反应（PCR）来检测。最近有报道称，通过对B细胞恶性肿瘤患者血浆中提取的DNA进行PCR分析，可以证实克隆细胞的增殖。然而，问题仍然是，扩增产物是否来自肿瘤细胞。在本研究中，我利用从同一B细胞白血病患者的恶性细胞和血浆中提取的DNA，尝试用半巢式PCR扩增免疫球蛋白重链（IgH）互补决定区3(CDR3)，并分别对其序列进行分析。结果表明，在所有病例中，血浆DNA序列与肿瘤细胞DNA序列相同。此外，其中一些与先前报道的CDR3具有同源性。这些发现表明血浆DNA是从肿瘤细胞中释放出来的。在下一阶段，有必要提高灵敏度，如果成功，使用血浆DNA的IgH-PCR可能成为诊断患者和评估治疗效果的有用工具，特别是在难以进行活检的危重患者中。

项目成果

Suzuki R, Ohtake S, et al.: "Prognostic significance of CD7+CD56+ phenotype and chromosome 5 abnormalities for acute myeloid leukemia MO."Int J Hematol.. 77. 482-489 (2003)

Suzuki R、Ohtake S 等人：“CD7 CD56 表型和 5 号染色体异常对急性髓系白血病 MO 的预后意义。”Int J Hematol.. 77. 482-489 (2003)

Ohtake S: "Chronic myelogenous leukemia with p190BCR-ABL expression : the missing link with monocytosis"Intern Med. 41. 1092-1093 (2002)

Ohtake S：“具有 p190BCR-ABL 表达的慢性粒细胞白血病：与单核细胞增多症缺失的环节”Intern Med。

Ohtake S: "Chronic myelogenous leukemia with p19OBCR-ABL expression : the missing link with monocytosis."Intern Med.. 41(12). 1092-1093 (2002)

Ohtake S：“具有 p19OBCR-ABL 表达的慢性粒细胞白血病：与单核细胞增多症缺失的环节。”Intern Med.. 41(12)。

Zhu J, Okumura H, Ohtake S, et al.: "Arsenic trioxide induces apoptosis in leukemia/lymphoma cell lines via the CD95/CD95L system."Oncol Rep.. 10(3). 705-709 (2003)

Zhu J、Okumura H、Ohtake S 等人：“三氧化二砷通过 CD95/CD95L 系统诱导白血病/淋巴瘤细胞系凋亡。”Oncol Rep.. 10(3)。

大竹茂樹: "急性骨髄性白血病"臨床医. 28・10. 2107-2110 (2002)

大竹茂树：“急性髓系白血病”临床医生。28・10（2002）。