Basic study for establishment of chemotherapy of DNA mismatch repair-deficient colorectal cancers : Specificity of DNA damage and drug sensitivity

DNA错配修复缺陷型结直肠癌化疗方案建立的基础研究：DNA损伤的特异性和药物敏感性

• 批准号: 12671273
• 负责人: FUNAHASHI Kimihiko
• 金额: $ 2.24万
• 依托单位: TOHO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671273/
• 关键词: colorectal cancer; DNA mismatch repair genes; hMLH1 gene; hMSH2 gene; drug-resistance; chemotherapeutic agents; cell lines; ミスマッチ修復遺伝子

Accumulation of mutations in oncogenes and tumor suppressor genes is esential for generation of colorectal cancer. Defect of a DNA mismatch repair (MMR) gene is known to be responding to one of the accumulated mutations in these cancer-related genes. Seven human MMR genes including hMLH1 and hMSH2 have been identified. MMR-deficient tumor cells exhibit a drug-resistance to some anticancer agents such as cisplatin (CDDP). However, sensitivities of MMR-deficient cells to many of anticancer drugs, which are utilizing in chemotherapy for colorectal cancers, and agents that induce MMR-tumor cells are not clear. We investigated here sensitivity of MMR-deficient colorectal cancer cell lines to the anticancer agents and a physiological role of the MMR system as follows :1) Drug-sensitivity of MMR-deficient cellsCytotoxicities of 6 anticancer agents in the MMR-deficient HCT116 and LoVo cell lines were compared to those of paired MMR-proficient cells, HCT116+ch3 and LoVo+ch2. HCT116 exhibited re … More sistance to 5-FU, tegafur, and irinotecan, while cytotoxicity of gemicitabine in HCT116 and LoVo was greater than that of MMR-proficient cells.2) Mechanism of high sensitivity of MMR-deficient cellsTo explore why MMR-deficient cells show high sensitivity, we had several experiments with aphidicolin and hydroxurea. The results strongly indicate that MMR system is closely involved in the DNA replication completion checkpoint, though further investigation is required.3) Establishment of a rapid and easy method for cytotoxic assay.To estimate how different cytotoxicity of anticancer agents between MMR-deficient and -proficient cells, we developed a rapid assay system using E. coli with expression plasmids.4) Presence of MMR-protein in tumor tissues of patients.To know the contribution of chemotherapeutic agents to appearance of MMR-deficient cells, we screened more than 100 specimens by immunohistochemical staining. Several samples was negative for the staining, though all cases were primary tumor. Less

癌基因和抑癌基因突变的积累是大肠癌发生的必要条件。已知DNA错配修复（MMR）基因的缺陷是对这些癌症相关基因中累积突变之一的反应。已鉴定出包括hMLH 1和hMSH 2在内的7个人类MMR基因。MMR缺陷型肿瘤细胞对一些抗癌药物如顺铂（CDDP）表现出耐药性。然而，MMR缺陷细胞对许多用于结肠直肠癌化疗的抗癌药物以及诱导MMR肿瘤细胞的药物的敏感性尚不清楚。我们研究了MMR缺陷型结直肠癌细胞系对抗癌药物的敏感性以及MMR系统的生理作用，具体如下：1）MMR缺陷型细胞的药物敏感性比较了6种抗癌药物对MMR缺陷型HCT 116和LoVo细胞系以及成对的MMR活性细胞HCT 116 + ch 3和LoVo+ ch 2的细胞毒性。HCT 116展示了re ...更多信息 对5-FU、替加氟和伊立替康的耐受性，而吉西他滨在HCT 116和LoVo中的细胞毒性大于MMR-熟练细胞。2）MMR-缺陷细胞的高敏感性的机制为了探索MMR-缺陷细胞显示高敏感性的原因，我们用阿非迪霉素和羟基脲进行了几个实验。结果表明，MMR系统与DNA复制完成检查点密切相关，但仍需进一步研究。3）建立一种快速、简便的细胞毒检测方法。4）患者肿瘤组织中MMR蛋白的存在为了解化疗药物对MMR缺陷细胞的影响，我们对100多例患者肿瘤组织进行了免疫组化筛选。尽管所有病例均为原发性肿瘤，但几个样本的染色呈阴性。少

项目成果

Zhong X.: "A single nucleotide polymorphism in the promoter regin of the hMLH1 gene : Effect on transcriptional activity and application for a marker of detecting allelic loss"J Med Soc Toho. 48・2. 114-124 (2001)

钟X.：“hMLH1基因启动子区的单核苷酸多态性：对转录活性的影响以及检测等位基因丢失的标记的应用”J Med Soc Toho 48·2（2001）。

Yoshikawa K: "Abnormal expression of BRCA1 and BRCA1-interactive DNA-repair proteins in breast carcinomas."Int J Cancer. 88(1). 28-36 (2000)

Yoshikawa K：“乳腺癌中 BRCA1 和 BRCA1 相互作用 DNA 修复蛋白的异常表达。”Int J Cancer。

Watanabe Y: "Complementation of an hMSH2-defect in human colorectal carcinoma cells by human chromosome 2 transfer."Mol Carcinog. 29(1). 37-49 (2000)

Watanabe Y：“通过人类 2 号染色体转移来补充人类结直肠癌细胞中的 hMSH2 缺陷。”Mol Carcinog。

Iwasaki I: "Establishment of cisplatin-resistant variants of human neuroblastoma cell lines, TGW and GOTO, and their drug cross-resistance profiles"Cancer Chemother Pharmacol. (in press). (2002)

Iwasaki I：“人神经母细胞瘤细胞系 TGW 和 GOTO 的顺铂耐药变体的建立及其药物交叉耐药性概况”Cancer Chemother Pharmacol。

鈴木康司: "大腸癌肝転移切除後予防的肝動注療法の臨床的意義"日本消化器外科学雑誌. 33. 169-175 (2000)

Koji Suzuki：“结直肠癌肝转移切除术后预防性肝动脉输注治疗的临床意义”日本胃肠外科杂志 33. 169-175 (2000)。