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Application of HiCEP, a novel method which can detect known and unknown gene expression changes after exposure to low dse X-rays
https://repo.qst.go.jp/records/61516
https://repo.qst.go.jp/records/615166fad1b29-a9ee-4d20-8463-8a302edde24a
| Item type | 会議発表用資料 / Presentation(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2006-06-26 | |||||
| タイトル | ||||||
| タイトル | Application of HiCEP, a novel method which can detect known and unknown gene expression changes after exposure to low dse X-rays | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_c94f | |||||
| 資源タイプ | conference object | |||||
| アクセス権 | ||||||
| アクセス権 | metadata only access | |||||
| アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
| 著者 |
Okayasu, Ryuichi
× Okayasu, Ryuichi× Fujimori, Akira× Takahashi, Sentaro× 岡安 隆一× 藤森 亮× 高橋 千太郎 |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | Application of HiCEP, a novel method which can detect known and unknown gene expression changes after exposure to low dose x-rays \nRyuichi Okayasu, Akira Fujimori, and Sentaro Takahashi \nResearch Center for Charged Particle Therapy Heavy-Ion Radiobiology Research Group National Institute of Radiological Sciences Anagawa 4-9-1, Inage-ku, Chiba 263-8555, Japan (E-mail, rokayasu@nirs.go.jp) Keywords: HiCEP, gene expression, low dose, CXC chemokines, normal human fibroblasts \nAlthough one can observe rapid advances in gene expression profiling method, it still is not easy to access the biological effects of radiation doses at the level of diagnostic radiology. In order to obtain better sensitivity, we used a novel gene expression profiling method, HiCEP (high coverage gene expression profiling) (Nucleic Acids Res. 31, e94) recently developed in our institute. HiCEP assay is an AFLP-based profiling method and has seven basic steps which include 1) double strand cDNA (dscDNA) synthesis with biotinylated oligo(dT) primer, 2) Digestion of dscDNA by MspI (or MseI), 3) Ligation with MspI (MseI) adapter and purification by magnetic beads coated with avidin, 4) Digestion of dscDNA by MseI (or MspI), 5) Ligation with MseI (or MspI) adapter, 6) Selective PCR with 256 primers (16X16), 7) Detection of the fluorescence from selective PCR products. These products were analyzed by capillary electrophoresis. In data profile, the x-axis indicates the length of PCR products and the y-axis indicates the intensity of fluorescence of the PCR products. This assay is highly reproducible and sensitive enough to detect 1.2-fold difference in gene expression. Moreover, the low false positive rate enables us to analyze gene expression with wide coverage. It is estimated that 70-80% of all transcripts which include non-coding transcripts and unknown and known genes. Using HiCEP, we searched through over 23,000 transcripts in normal human fibroblasts (HFL III) irradiated with low dose X-rays. More than 200 genes are up-regulated transiently during one hour after 1 cGy X-rays. We determined the nucleotide sequences of 10 up-regulated transcripts with the greatest rate of increase. Three out of these encoded a set of CXC chemokine genes (CXCL1, CXCL2, and CXCL6). The rest included the transcripts of other secretory products and unknown genes. In order to test the involvement of CXC chemokines in cells irradiated with low doses, we irradiated HFL III cells with 1-10 cGy X-rays, and transferred the media from HFL III culture to two melanoma cell lines characteristic of excessive numbers of the CXC chemokine-specific receptors. The growth of these melanoma lines were significantly stimulated after the addition of HFL III medium irradiated at 1-5 cGy (Cancer Res. 65, 10159-10163, 2005). These results suggest an existence of a defense mechanism specific for exposure to low doses of ionizing radiation. |
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| 会議概要（会議名, 開催地, 会期, 主催者等） | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | The sixth Japan-France workshop on radiobiology and isotopic imaging | |||||
| 発表年月日 | ||||||
| 日付 | 2006-06-22 | |||||
| 日付タイプ | Issued | |||||
