Establishment of a microdetection method for DNA mismatch repair deficiency in colorectal cancers and its clinicopathological study

结直肠癌DNA错配修复缺陷显微检测方法的建立及临床病理研究

• 批准号: 10671222
• 负责人: TSUJITA Kazunori
• 金额: $ 2.37万
• 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: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671222/
• 关键词: colorectal cancer; anti-hMLH1 antibody; anti-hMSH2 antibody; microsatellite instability; gene abnormality; 簡易検索法; 免疫組織化学的染色法; 臨床病理学的要因; hMLH1蛋白; hMSH2蛋白; RT-PCR; 臨床病理学要因

DNA mismatch repair (MMR) genes are responsible to hereditary nonpolyposis colorectal cancer and some of sporadic colorectal cancers. MMR-deficient tumor cells exhibit a low level of resistance to some anti-cancer agents, such as cisplatin, adriamycin, and alkylating agents. To know the MMR status or microsatellite instability (MSI) status of tumor cells is critical for choosing a therapeutical protocol. In this study, we established methods for microdetection of hMSH2 and hMLH1 proteins with monoclonal antibodies and MSI status determination. Further, clinicopathologic features of 99 cases of sporadic colorectal cancer was investigated.(1)MSI statusWe selected 16 microsatellite markers and determined an efficient detection procedure for MSI status by studying in 90 cases of sporadic colorectal cancer.(2)Expression of mismatch repair proteins, hMSH2 and hMLH1Expression of hMSH2 and hMLH1 proteins in 99 samples of sporadic colorectal cancer selected randomly was analyzes by Western blot … More and immunohistochemical staining with monoclonal antibodies developed in our laboratory. Totally 10 tumor samples with no/little expressing hMSH2 (4 samples) and hMLH1(6 samples) proteins were found, indicating approximately 10% of sporadic cases and 67% of the MSI tumor have some defects in the expression of these MMR proteins.(3)Mutations in hMSH2 and hMLH1 genesTo know the molecular mechanism of the suppression of the gene expression, mutation in the exons were studied. Two kinds of mutations, which affect protein function, in a hMSH2-deficient sporadic case and one non-sense mutation in hMSH2-deficient a familial case were found. No mutation was found in hMLH1-deficient cases.(4)Clinicopathologic featuresClinicopathologic features of 17 cases of MSI tumor showed similar characteristics to those frequently found in HNPCC cases such as right-sided location of the primary tumor, a lower frequency of well-differentiated adenocarcinoma, and Duke's staging A or B.Further study is required to clarify the molecular mechanism of suppression of MMR gene expression, especially hMLH1 gene. In addition, since MMR-deficient or MSI tumors show a low level of drug-resistance, the specified protocol for treatment of MSI tumor should be developed. Less

DNA不匹配修复（MMR）基因是遗传性非型结直肠癌和一些零星结直肠癌的原因。缺乏MMR的肿瘤细胞表现出对某些抗癌剂的耐药性，例如顺铂，阿霉素和烷基化剂。要了解肿瘤细胞的MMR状态或微卫星不稳定性（MSI）状态对于选择治疗方案至关重要。在这项研究中，我们建立了具有单克隆抗体和MSI状态测定的HMSH2和HMLH1蛋白进行微调的方法。 Further, clinicopathological features of 99 cases of sporadic colorectal cancer was investigated.(1)MSI statusWe selected 16 microsatellite markers and determined an efficient detection procedure for MSI status by studying in 90 cases of sporadic colorectal cancer.(2)Expression of mismatch repair proteins, hMSH2 and hMLH1Expression of hMSH2 and hMLH1 proteins in 99 samples of通过蛋白质印迹对随机选择的螺旋结直肠癌进行了分析……更多的和免疫组织化学染色，并用我们实验室中开发的单克隆抗体进行了染色。发现完全10个肿瘤样品HMSH2（4个样品）和HMLH1（6个样品）蛋白，表明在这些MMR蛋白的表达中，大约有10％的螺旋病例和67％的MSI肿瘤在HMSH2和HMLH1纯料中的表达中存在一些缺陷。 Studiod。在HMSH2缺乏零星的情况下，发现了两种影响蛋白质功能的突变，在HMSH2缺乏症中发现了一种养殖情况。 No mutation was found in hMLH1-deficiency cases.(4) Clinicopathological features Clinicopathological features of 17 cases of MSI tumor shown similar characteristics to those frequently found in HNPCC cases such as right-sided location of the primary tumor, a lower frequency of well-differentiated adenocarcinoma, and Duke's staging A or B.Further study is required to clarify the molecular mechanism of suppression of MMR gene表达，尤其是HMLH1基因。此外，由于含MMR或MSI肿瘤的药物耐药水平较低，因此应制定用于治疗MSI肿瘤的指定方案。较少的

项目成果

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船橋 公彦: "大腸癌先進部における組織型とki-67標識率の検討" 日本消化器外科学会雑誌. 58. 846-852 (1998)

Kimihiko Funabashi：“结直肠癌晚期区域的组织类型和 KI-67 标记率检查”日本胃肠外科学会杂志 58. 846-852 (1998)。