細胞障害により惹起される分子指標を用いた新らしい抗癌剤感受性試験

利用细胞损伤诱导的分子指标进行新的抗癌药物敏感性测试

• 批准号: 15790703
• 负责人: 赤津 友佳子
• 金额: $ 1.34万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15790703/
• 关键词: chemosensitivity test; gastric cancer; 5-fluorouracil; Cisplatin; gadd15; p21; c-Jun; 抗癌剤感受性試験; 遺伝子発現; 早期判定

【目的】近年,進行胃癌に対し新規抗癌剤の効果が期待される一方,これらの抗癌剤に対するnon-responderも存在し,こうした症例の早期発見が重要である。我々は、癌化学療法の効果予測を治療早期に予測しうるassayの開発を目的として検討を行った。【方法と対象】化学療法開始早期に細胞死に先立ち変化を来たす遺伝子の候補としてgadd153、p21,c-junを採用した。(in vitro)ヒト胃癌細胞株TMK-1,MKN-45,MKN-74を,低〜高濃度5-FU,CDDPにそれぞれ曝露後,経時的に各遺伝子のmRNAの発現を定量した。抗腫瘍効果はMTT assayを用いて判定し、無治療群を対照群とした.(in vivo)ヒト胃癌細胞株TMK-1をnude mouseに皮下移植し腫瘍を作成した後,低〜高濃度5-FU,CDDPを腹腔内投与した。薬剤投与後48時間に皮下腫瘍を回収しm-RNAを同様に定量した。また、それぞれの薬剤濃度による抗腫瘍効果は薬剤投与後21日目に評価した。(臨床例)高度進行胃癌患者11人に対し化学療法施行前と、施行後1週間以内に胃内視鏡下生検にて癌組織を採取、m-RNAを定量し、治療効果との関係を検討した【結果】(in vitro)全ての細胞株において,用量,曝露時間に比例し癌細胞増殖阻害を認め、遺伝子発現を強く認めた。細胞間、薬剤間にmRNA発現量に差を認めず、抗腫瘍効果を予測するcut off値をROC曲線からgaddl53 1.3、p21 1.8、c-Jun2.1と算定しえた。 (in vivo)in vitroの実験と同様に、腫瘍増殖抑制を認めた高濃度5-FU,CDDPで、薬剤投与後48時間での各mRNA発現増強を認め、同様にcut off値をROC曲線からgadd153 1.8、p21 1.9、c-Jun2.2と算定しえた。(臨床例)治療開始前後のmRNA発現量は、PRの症例がPDの症例より増加傾向にあった。【結語】化学療法開始後早期の遺伝子発現量は抗腫瘍効果を反映する可能性が示唆された。

[Objective] In recent years, new regulations for gastric cancer have been carried out, and the results of anti-cancer agents are expected. However, the existence of non-responders for anti-cancer agents is important, and early detection of cases is important. We are looking forward to early detection of cancer chemotherapy outcomes and development of assay objectives. [Methods] Early chemotherapy begins, cell death begins, and the candidate for the gene is gadd153, p21,c-jun. (in After exposure to low to high concentrations of 5-FU and CDDP, mRNA expression of each gene was quantified in gastric cancer cell lines TMK-1,MKN-45,MKN-74. Anti-tumor effect MTT assay is used to determine whether there is a treatment group or not. (in After subcutaneous transplantation of TMK-1 gastric cancer cell line into nude mice, low to high concentrations of 5-FU and CDDP were administered intraperitoneally. Subcutaneous lesions were recovered 48 hours after administration and m-RNA was quantified simultaneously. The anti-tumor effect of the drug was evaluated 21 days after administration. 11 highly advanced gastric cancer patients were examined under gastroscopy before and within 1 week after chemotherapy for the relationship between tissue sampling, m-RNA quantification, and therapeutic effects.[Results] All cell lines were identified in vitro for inhibition of cancer cell proliferation, and for the proportion of exposure time. The ROC curve for the difference in mRNA expression between cells and tissues was calculated from gaddl53 1.3, p21 1.8 and c-Jun2.1. (in In vivo)in vitro, the expression of each mRNA increased at 48 hours after 5-FU,CDDP, and drug administration, and ROC curves were determined by gadd153 1.8, p21 1.9, and c-Jun2.2. (Clinical cases) mRNA expression before and after treatment, PR cases and PD cases increased. Conclusion: Early gene expression after initiation of chemotherapy may reflect the likelihood of anti-tumor effects.