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DNAマイクロアレイとLCM法を用いた口腔癌関連遺伝子データベースの構築

利用DNA微阵列和LCM方法构建口腔癌相关基因数据库

基本信息

批准号：
14771159
负责人：
羽鳥仁志
金额：
$ 2.5万
依托单位：
Showa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
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/grant/KAKENHI-PROJECT-14771159/
关键词：
LCM IMS Gene Chip DNAマイクロアレイ口腔癌関連遺伝子

项目摘要

【平成15年度】同意の得られた口腔癌患者10例の手術時検体(原発巣)から、Cell Robotics社製のCRI-337を用いたLaser Captured Microdissection(LCM)法にて約200個の癌細胞のみを切り出し、Total RNAを抽出した。また、同一患者の末梢血5mlを採取し、抗上皮抗体磁気ビーズを用いたImmunomagnetic separation(IMS)法により、末梢血中微小転移癌細胞を補足し、同様にRNAを抽出した。これら2つの方法にて回収したRNAを試料とし、RT-PCR法にて転移関連遺伝子群の発現パターンを解析し、比較検討を行った。その結果、細胞外基質分解酵素群のうちマトリックスメタロプロテアーゼ(MMP)-1,2,7,9は、腫瘍原発巣での発現が多く認められる傾向にあり、細胞接着因子CD44は、そのスプライスバリアント(v6,v9>を含め原発巣ではほとんど発現を認めず、末梢血サンプルで高頻度に認められた。一方、同一患者の腫瘍原発巣から回収した試料を用い、Gene Chip発現解析システム(Affimetrix)による癌関連遺伝子群の包括的発現パターンの解析を行うための予備実験、すなわちマウスTotal RNAを試料とした種特異的遺伝子発現パターンのGene Chip Test Arrayによる解析は終了しており、臨床材料の検討を始めている。現在、これら癌関連遺伝子群の発現パターンと、各症例の臨床・病理分類、リンパ節転移様式を比較検討し、データベースを作成中である。

[Pingcheng 15] agreed that 10 patients with oral cancer underwent manual surgery, and the Cell Robotics community CRI-337 was used to evaluate about 200 cancer cells by the method of Laser Captured Microdissection (LCM), and Total RNA was extracted. 5ml was extracted from the peripheral blood of the same patient, anti-epithelial antibody magnetic antibody was detected by Immunomagnetic separation (IMS) method, micrometastatic cancer cells were extracted from the peripheral blood and extracted from the same patient. In this paper, the RNA method is used to analyze the data, and the RT-PCR method is used to move the link subgroup to analyze and compare the data. The results showed that the extracellular motif decomposing enzyme group (MMP)-1 was found to be highly sensitive (v6 ~ 9), and the cell-linked factor CD44 (v6 ~ (9) g), which contained the original enzyme, was found to be highly sensitive and sensitive. One party, the same patient, did not know that the material was used, and Gene Chip found that the subgroup of the Affimetrix link to cancer, including the subgroup of the link between cancer and cancer, showed that the subgroup of the link between cancer and cancer, including the subgroup of the link between cancer and cancer, showed that the subgroup of the sub-group of the link between cancer and cancer, which was included in the sub-group of the link between cancer and cancer, and that the subgroup of the link, which was included in the subgroup of the link between cancer and cancer, and that the sub-group of the patient and the patient of the same patient, was not in the first place, that the data of the Gene Chip Test Array was analyzed in the first place, and that the material of the bed was analyzed. At present, in the subgroup of cancer patients, we can see that the clinical symptoms, the pathological classification of each case, the transfer type of the disease, and the diagnosis of the disease are in progress.