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预后不良密切相关。虽然N-myc扩增在神经母细胞瘤中的检测具有重要意义，但目前单独使用的Southern杂交存在耗时、需要大量DNA和操作复杂等缺点。为了克服这些缺点，我们计划将竞争性PCR（cPCR）应用于N-myc扩增的检测。1995年，我们建立了利用竞争质粒pZH 2测定N-myc扩增的cPCR体系，1996年，我们又用pZH 2作为竞争质粒，建立了一种新的检测N-myc扩增的cPCR体系，并在此基础上建立了一种新的检测N-myc扩增的cPCR方法。我们使用少量样品检测了cPCR在拷贝数测定中的可用性。我们检测了6例神经母细胞瘤穿刺活检标本的N-myc拷贝数，并证明了其可用性。综上所述，本研究建立的cPCR方法是一种快速、简便、适用于小样本的N-myc拷贝数检测方法，可作为Southern杂交检测N-myc拷贝数的替代方法。

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）。