Development of the molecular diagnostic tests for drug-resistant cancer cells based on gene expression profiling using microarray

基于使用微阵列的基因表达谱开发耐药癌细胞的分子诊断测试

• 批准号: 17590499
• 负责人: MIYACHI Hayato
• 金额: $ 2.24万
• 依托单位: Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590499/
• 关键词: anti-cancer agents; DNA maicroarray; gene expression array; leukemia

Drug resistance is a major obstacle in chemotherapy of leukemia patients. Since the treatment responsiveness is different among patients, the therapy should be tailored to specific subtypes of diseases such as defined by drug resistance. We attempted to develop the molecular diagnostic tests for drug-resistant cancer cells based on gene expression profiling using a microarray method. The aim of this study was to select a gene panel for a diagnostic array by analyzing function of candidate genes in the drug resistance. The ATM gene was chosen as a novel and candidate gene. ATM was found overexpressed in trimetrexate-resistant leukemia MOLT-3 cells (MOLT-3/TMQ200) as analyzed by gene expression profiling using a commercially available cDNA microarray (Human Cancer CHIP version 3.0, TaKaRa Biotechnology). It was confirmed by Northern blot analysis. When the plasmid with insertion of the siRNA for ATM was transfected to MOLT-3/TMQ200 cells, ATM expression was decreased as analyzed by RT-PCR. Expression levels of ATM as assessed by real-time PCR was increased in parallel with drug resistance degree in three sublines obtained from MOLT-3/TMQ cells. Resistance to irradiation as assessed by GO values in agarose clonogenic assay was increased in parallel with the ATM levels in three sublines. The novel gene which was demonstrated as functioning in the mechanisms of drug resistance would be a useful marker for molecular diagnostic tests for drug-resistant cancer cells based on gene expression profiling using a microarray method.

耐药是白血病患者化疗的主要障碍。由于不同患者的治疗反应不同，治疗应针对特定的亚型疾病，如耐药定义。我们尝试开发基于微阵列方法的基因表达谱的耐药癌细胞的分子诊断测试。本研究的目的是通过分析候选基因在耐药中的功能来选择用于诊断阵列的基因面板。ATM基因被选为一个新的候选基因。通过使用商用的基因芯片芯片(人类癌症芯片3.0版，Takara Biotech)进行基因表达谱分析，发现ATM在曲美他克星耐药白血病MOLT-3细胞(MOLT-3/TMQ200)中过表达。Northern印迹分析证实了这一点。将插入ATM的siRNA导入MOLT-3/TMQ200细胞后，RT-PCR结果显示ATM的表达明显降低。在MOLT-3/TMQ细胞中，ATM的表达水平随着耐药程度的增加而增加。根据琼脂糖克隆形成实验中的GO值评估的辐射抗性在三个亚系中与ATM水平平行地增加。该新基因被证明在耐药机制中发挥作用，将成为基于微阵列方法基因表达谱的耐药癌细胞分子诊断测试的有用标记。

项目成果

Treatment of multi-drug resistant acute leukemia by cleavage of MDR1 mRNA with DNA enzyme.

通过 DNA 酶切割 MDR1 mRNA 治疗多重耐药急性白血病。

• 发表时间: 2005
• 期刊: Rec.Adv.Clin.Pharmacol 2005
• 作者: Asai S;Kobayashi H;Mivachi H.
• 通讯作者: Mivachi H.

In vivo analysis of the role of aberrant hitone deacetylase recruitment and RAR α blockade in the pathogenesis of acute promyelocytic leukemia.

体内分析异常蛋白脱乙酰酶募集和 RAR α 阻断在急性早幼粒细胞白血病发病机制中的作用。

• 发表时间: 2006
• 期刊: J. Exp. Med. 203:4
• 作者: Matsushita H;Scaglioni PP;Bhaumik M;Rego EM;Lu FC;Majid SM;Miyachi H;Kakizuka A;Miller Jr WH;Pandolfi PP
• 通讯作者: Pandolfi PP

MDR1mRNA特異的切断活性をもつDNAenzymeによる多剤耐性ヒト急性白血病の治療.

使用具有 MDR1 mRNA 特异性切割活性的 DNA 酶治疗多重耐药人类急性白血病。

• 发表时间: 2005
• 期刊: 臨床薬理の進歩 2005
• 作者: 浅井さとみ;小林広幸;宮地勇人
• 通讯作者: 宮地勇人