Establishment of quantitative RT-PCR method to defect drug-resistant-related genes

耐药相关基因缺陷定量RT-PCR方法的建立

• 批准号: 09672348
• 负责人: FUNATO Tadao
• 金额: $ 2.37万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672348/
• 关键词: drug resistance; leukemia; real-time RT-PCR; Multidrug reistance; cytosine arabinoside; 多剤耐性遺伝子(MDR); 薬剤耐性遺伝子; 遺伝子診断

The failure of chemotherapy to eradicate human leukemic cells is often due to the development of drug resistance. MDR (multidrug resistance) whose one form of drug resistance results from a decreased intracellular accumulation of the drugs, most often mediated by the overexpression of P-glycoprotein. Multidrug-related protein (MRP) also related to pump function of cell membrane in acute leukemia. Decreased deoxycytidine kinase (dCK) gene was observed in cytosine arabinoside resistant leukemia cells (ara-C) which we have established. We have developed the new quantitative assay based on real-time reverse transcriptase polymerase chain reaction (RT-PCR) to mesure expression of drug-resistance related genes such as MDR-1, MRP, and dCK in clinical samples. These results indicates that real-time RT-PCR system is a releable method to quantitatively determine drug resistant genes expression, it may be to predict responsiveness to chemotherapy by using this technique.

化疗未能根除人类白血病细胞通常是由于耐药性的产生。 MDR（多药耐药性），其一种形式的耐药性是由于细胞内药物积累减少造成的，通常是由 P-糖蛋白的过度表达介导的。多药相关蛋白（MRP）也与急性白血病细胞膜的泵功能有关。在我们建立的胞嘧啶阿糖苷抗性白血病细胞（ara-C）中观察到脱氧胞苷激酶（dCK）基因减少。我们开发了基于实时逆转录聚合酶链式反应 (RT-PCR) 的新定量检测方法，用于测量临床样本中 MDR-1、MRP 和 dCK 等耐药相关基因的表达。这些结果表明实时RT-PCR系统是定量测定耐药基因表达的可靠方法，利用该技术可以预测对化疗的反应。

项目成果

舩渡 忠男,他: "がんの薬剤耐性の遺伝子診断"臨床病理. 48. 162-166 (2000)

Tadao Funato 等：“癌症耐药性的基因诊断”《临床病理学》48. 162-166 (2000)。

Funato T.et al.: "In vitro leukemia cell models of ara-C resistance"Leukemia Research. (in press).

Funato T.等人：“ara-C 耐药性的体外白血病细胞模型”白血病研究。

舩渡 忠男、他: "定量的PCRシステムの臨床応用" 臨床病理. 46・5. 339-405 (1998)

Tadao Funato等：“定量PCR系统的临床应用”临床病理学46・5（1998）。

籐巻 慎一,他: "簡便な定量的PCRシステムによる白血病MRDの遺伝子診断"International J Haemtol. 69suppl. 67 (1999)

Shinichi Satomaki 等人：“使用简单定量 PCR 系统进行白血病 MRD 的基因诊断”International J Haemtol 69 (1999)。

籐巻 慎一,他: "定量RT-PCRにおけるRNA基準量の設定に関する検査"臨床病理. 48. 54-59 (2000)

Shinichi Satomaki 等人：“定量 RT-PCR 中设置 RNA 参考量的测试”《临床病理学》48. 54-59 (2000)。