p57KIP2 基因异常甲基化是检测弥漫性大 B 细胞淋巴瘤微小残留病的敏感生物标志物。

Aberrant DNA methylation of the p57KIP2 gene is a sensitive biomarker for detecting minimal residual disease in diffuse large B cell lymphoma.

机构信息

Clinical Research Centre, National Hospital Organization, Nagoya Medical Center, Naka-ku, Nagoya, Japan.

出版信息

Leuk Res. 2010 Jan;34(1):50-4. doi: 10.1016/j.leukres.2009.06.028. Epub 2009 Jul 18.

DOI:10.1016/j.leukres.2009.06.028

PMID:19616848

Abstract

The detection of minimal residual disease (MRD) in bone marrow is very important in the clinical management of malignant lymphoma. So far, the assessment of MRD in cases of diffuse large B cell lymphoma (DLBCL) has had some technical limitations, such as requiring patient-specific primers and complicated experimental steps. To resolve these problems, we applied a tumor-specific epigenetic alteration of the p57KIP2 gene as a biomarker for detecting MRD in DLBCL. The methylation of the p57KIP2 gene was analyzed in 63 cases of DLBCL by methylation-specific real-time quantitative PCR. Methylation of the p57KIP2 gene was detected in 53 (84.1%) of these 63 cases of DLBCL. We could detect one p57KIP2 gene-methylated cell among 10,000 unmethylated cells by the serial dilution experiment. This sensitivity is proved to be equivalent to that of detection of bcl2/IgH rearrangement by real-time quantitative PCR. This sensitivity could be converted to the detection of two methylated genomes per reaction. Using clinical material, the same results were confirmed. In this study, we established a convenient and universal method for detecting MRD in DLBCL. This technique is applicable for over 80% of patients with DLBCL. This could promote systemic MRD studies in the area of DLBCL.

摘要

骨髓微小残留病（MRD）的检测在恶性淋巴瘤的临床管理中非常重要。到目前为止，弥漫性大 B 细胞淋巴瘤（DLBCL）的 MRD 评估存在一些技术限制，例如需要患者特异性引物和复杂的实验步骤。为了解决这些问题，我们应用了 p57KIP2 基因的肿瘤特异性表观遗传改变作为检测 DLBCL 中 MRD 的生物标志物。通过甲基化特异性实时定量 PCR 分析了 63 例 DLBCL 中 p57KIP2 基因的甲基化。我们在 63 例 DLBCL 中检测到 53 例（84.1%）p57KIP2 基因甲基化。通过系列稀释实验，我们可以在 10000 个未甲基化细胞中检测到一个 p57KIP2 基因甲基化细胞。这种敏感性被证明与实时定量 PCR 检测 bcl2/IgH 重排相当。这种敏感性可以转化为每个反应检测两个甲基化基因组。使用临床材料，也证实了相同的结果。在这项研究中，我们建立了一种方便和通用的方法来检测 DLBCL 中的 MRD。该技术适用于 80%以上的 DLBCL 患者。这将促进 DLBCL 领域的系统性 MRD 研究。