Establishment of a competitive PCR for N-myc amplification in neuroblastoma

神经母细胞瘤 N-myc 扩增竞争性 PCR 的建立

• 批准号: 07671958
• 负责人: INOUE Akira
• 金额: $ 1.41万
• 依托单位: Toho University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671958/
• 关键词: neuroblastoma; N-myc; gene amplification; PCR; competitive PCR; prognosis; N-myc遺伝子; 小児固型腫瘍

N-myc amplification correlates well with the poor prognosis of neuroblastoma. Although the detection of N-myc amplification in neuroblastoma is of great importance, Southern hybridization, being employed solely so far, has some disadvantages such as time consuming, the requirement of larger amount of DNA,and somewhat complicated procedure. In order to overcome the disadvantages, we planned to apply competitive PCR (cPCR) to the measurement of N-myc amplification. Although the standard polymerase chain reaction (PCR) is essentially unsuitable for the quantitative detection of a given template DNA,cPCR is an absolute determination of the dose of a given template DNA.In 1995, we established cPCR system for determining N-myc amplification by employing the competitor plasmid pZH2.In 1996, we examined the availability of cPCR in the copy-number determination using the samples with small amounts. We tested the N-myc copy number in 6 neuroblastoma samples collected by needle biopsy and demonstrated the availability. Taken together, we concluded cPCR established in this study is a rapid, simple, and being applicable to small samples and can be an alternate method of Southern hybridization to determine N-myc copy number.

N-MYC扩增与神经母细胞瘤的预后不良很好。尽管神经母细胞瘤中N-MYC扩增的检测非常重要，但到目前为止，南部杂交的杂交却具有一些缺点，例如耗时，更大量的DNA的需求以及有些复杂的程序。为了克服缺点，我们计划将竞争性PCR（CPCR）应用于N-MYC扩增的测量。 Although the standard polymerase chain reaction (PCR) is essentially unsuitable for the quantitative detection of a given template DNA,cPCR is an absolute determination of the dose of a given template DNA.In 1995, we established cPCR system for determining N-myc amplification by employing the competitor plasmid pZH2.In 1996, we examined the availability of cPCR in the copy-number determination using the samples with small金额。我们测试了通过针头活检收集的6个神经母细胞瘤样品中的N-MYC拷贝数，并证明了该副本的可用性。综上所述，我们得出的结论是在这项研究中建立的CPCR是一个快速，简单且适用于小样本的CPCR，可以是南方杂交的另一种方法来确定N-MYC拷贝数。

项目成果

Kong, X.-T., Choi, S.-H., Inoue, A., Takita, J., Yokota, J., Hanada, R., Yamamoto, K., Bessho, F., Yanagisawa, M., and Hayashi, Y.: "Genetic alterations of tumor suppressor gene DCC in neuroblastoma." European Journal of Cancer. (in press).

Kong，X.-T.，Choi，S.-H.，Inoue，A.，Takita，J.，Yokota，J.，Hanada，R.，Yamamoto，K.，Bessho，F.，Yanagisawa，M.

Inoue,A,et.al.: "Extensive genetic heterogeneity in a nouro blastoma cell line NB (TV) 1." International Journal of Cancer. (予定). (1997)

Inoue,A,et.al.：“努鲁母细胞瘤细胞系 NB (TV) 1 中的广泛遗传异质性。”国际癌症杂志（计划）。

Kong,x-T.,et al: "Genetic alterations of tumor suppressor gene DCC in neuro blastoma" European Journal of Cancer. (予定). (1997)

Kong, x-T., 等人：“神经母细胞瘤中肿瘤抑制基因 DCC 的遗传改变”《欧洲癌症杂志》（计划）（1997 年）。

Inoue,A.,et al.: "Extensive genetic heterogeneity in a neuroblastoma cell lire NB(TV)1" International Journal of Cancer. (予定). (1997)

Inoue, A. 等人：“神经母细胞瘤细胞 NB(TV)1 中的广泛遗传异质性”国际癌症杂志（计划）（1997 年）。

Akira Inoue: "Competitive Polymerase Chain Reaction for the Quantification of N-myc Gene Copy Number in Neuroblastoma" Tumor Biology. (発表予定). (1996)

Akira Inoue：“用于定量神经母细胞瘤中 N-myc 基因拷贝数的竞争性聚合酶链反应”肿瘤生物学（即将发表）。