成果報告書詳細
管理番号20140000000451
タイトル*平成25年度中間年報 「がん超早期診断・治療機器の総合研究開発 超早期高精度診断システムの研究開発:血液中のがん分子・遺伝子診断を実現するための技術・システムの研究開発 血中分子・遺伝子診断のための基礎技術の研究開発(がんの発症を予測するシステムの開発 乳がん感受性評価システムの研究開発)」
公開日2015/3/25
報告書年度2013 - 2013
委託先名国立大学法人山口大学
プロジェクト番号P10003
部署名バイオテクノロジー・医療技術部
和文要約
英文要約Research and development of a new system of risk assessment for breast cancer.

To develop a new system of risk assessment for breast cancer susceptibility, we have been performing this project.

Materials and Methods
We purchased 1,000 DNA samples from peripheral blood of women without a history of breast cancer and 1,000 DNA samples from peripheral blood of patients with a history of breast cancer from BioBank Japan in The University of Tokyo. We also obtained 214 DNA samples from peripheral blood of women without a history of breast cancer and 198 DNA samples from peripheral blood of patients with a history of breast cancer from Yamaguchi University Hospital. In addition, we collected 463 DNA samples for the prospective study from peripheral blood of women without a history of breast cancer.
Digital PCR was performed to evaluate absolute copy numbers. We designed forward and reverse primers and a TaqMan® MGB probe of target region and RNaseP. RNaseP was used as the internal control because it is known to exist in two copies in a diploid genome. Reaction mixtures of 20-μL volume comprising 1× ddPCR Master Mix (Bio-Rad), forward and reverse primers and probes for a target and a reference, and DNA were prepared. PCR amplification was performed for a total of 40 cycles with an annealing temperature of 56°C. Digital PCR was carried out using a QX100 droplet digital PCR system (BioRad) according to the manufacturer’s protocol.

Results
 Using 214 DNA samples from peripheral blood of women without a history of breast cancer and 198 DNA samples from peripheral blood of patients with a history of breast cancer from Yamaguchi University Hospital, we performed the digital PCR assay. The means of the relative copy numbers of women in the control group and those of patients with a history of breast cancer was 1.8 and 1.3 for Hs03898338_cn on 15q26.3 (P<0.0001) (Figure 1).
 
Conclusions
These findings may lead to a new means of risk assessment for breast cancer. Confirmatory studies are underway in the current project.
Figure 1. Distribution of copy numbers in women in the control group and in patients with a history of breast cancer and. Each sample is indicated by an open circle. The horizontal lines represent the mean copy number in each group.
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