Thymidylate Synthetase遺伝子発現より見た腫瘍の5-FU感受性の評価に関する研究

基于胸苷酸合成酶基因表达量评价肿瘤5-FU敏感性的研究

• 批准号: 08770971
• 负责人: 山口 由美
• 金额: $ 0.7万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-08770971/
• 关键词: thymidylate synthase; 5-FU; 化学療法感受性

腫瘍の5-FU感受性とthymidylate synthase(TS)遺伝子発現との関連を知るため以下のような実験を行った。1.胃癌細胞株MKN45,MKN74,MKN1,MKN28,KATOIIIの培養液中に5-FUを20mM,10mM,5mM,1mM,0.5mM.0.25mM,0.1mM加え、それぞれ6時間、24時間、4日間作用させ4日後の生細胞の率を測定した。その結果、(1)いずれの細胞株においても、5mM以上では、生細胞率は高濃度、長時間接触になるほど減少していた。(2)MKN45では0.5mM,MKN28では1mM,KATOIIIでは0.25mM,MKN74では0.1mM以下の濃度では、生細胞率は5-FUの濃度とは関係なく接触時間のみに依存してた。(3)比較的高い濃度で、生細胞率が接触時間依存性になる細胞株(MKN28,MKN45)では低濃度5-FUを作用させた際、5-FUに対する感受性が低い傾向にあった。2.上記の培養細胞株に5-FUを作用させ、20mM,10mM,5mMでは6時間後、1mM,0.5mM,0.25mM,0.1mMでは6時間、24時間、4日後にRNAを抽出しTS遺伝子の発現をPCRを用いて検討した。その結果、(1)5-FU投与前で、MKN28のTS遺伝子の発現が最も高く、次いで、MKN1,MKN45で発現が高くMKN74,KATOIIIではTS遺伝子の発現は低かった。5-FUを低濃度で用いた場合、TS遺伝子の発現が低い細胞株ほど感受性が高かった。しかし高濃度で用いた場合の感受性は、TS遺伝子発現との関連は認められなかった。(2)5-FU投与後のTS遺伝子の変化については現在検討中である。3.当科で胃癌、食道癌の手術を受ける患者に術前5-FUで化学療法を行い臨床効果、術前stageとTS遺伝子のレベルとの関連を検討中である。

5-FU susceptibility and thymidylate synthase(TS) gene expression in tumors 1. Gastric cancer cell lines MKN45, MKN74, MKN1, MKN28,KATOIII were cultured with 5-FU at concentrations of 20mM, 10mM, 5mM, 1mM, 0.5mM, 0.25mM, 0.1mM for 6 hours, 24 hours, 4 days and 4 days. The results are as follows: (1) The cell growth rate decreases when the concentration is higher than 5mM, and the cell growth rate decreases when the concentration is higher and the contact time is longer. (2) MKN45 <0.5mM, MKN28 <0.1mM,KATOIII <0.25mM, MKN74 <0.1mM, the cell growth rate <0.1mM and the concentration dependence of 5-FU. (3)Compared with high concentrations, cell growth rate was dependent on exposure time. Cell lines (MKN28, MKN45) tended to have low sensitivity to 5-FU when 5-FU acted on them at low concentrations. 2. RNA was extracted from the cultured cell lines after 6 hours, 24 hours and 4 days of 5-FU treatment at 20mM,10mM,5mM, 1 mM, 0.5 mM, 0.25 mM, 0.1 mM, and the expression of TS gene was detected by PCR. The results are as follows: (1)5-FU was injected before, MKN28 was injected before, MKN1 was injected after, MKN45 was injected after, MKN74 was injected after, KATOIII was injected after, MKN28 was injected before, MKN28 was injected after, MKN1 was injected after, MKN45 was injected after, MKN74 was injected after,KATOIII was injected after, MKN28 was injected after, MKN28 was injected after, MKN1 was injected after, MKN45 was injected after, MKN74 was injected after,KATOIII was injected after. 5-FU was used at low concentrations, and the expression of TS gene was low and the sensitivity of 5-FU was high. The sensitivity and correlation of TS gene expression in high concentration cases were analyzed. (2)5-FU and TS gene transformation after injection. 3. The relationship between preoperative 5-FU and chemotherapy in patients undergoing surgery for gastric cancer and esophageal cancer is discussed.