Kim Il-Jin, Kang Hio Chung, Park Jae-Hyun, Ku Ja-Lok, Lee Jong-Soo, Kwon Hyuk-Joon, Yoon Kyong-Ah, Heo Seung Chul, Yang Hee-Young, Cho Bo Youn, Kim Seong Yeon, Oh Seung Keun, Youn Yeo-Kyu, Park Do-Jun, Lee Myung-Shik, Lee Kwang-Woo, Park Jae-Gahb
Familial Cancer Clinic, National Cancer Center, 809 Madu-dong, Ilsan-gu, Goyang, Gyeonggi 411-764, Korea.
Clin Cancer Res. 2002 Feb;8(2):457-63.
Multiple endocrine neoplasia type 2 (MEN2) syndromes are inherited in an autosomal dominant fashion with high penetrance. There are three subtypes, namely, MEN2A (multiple endocrine neoplasia type 2A), MEN2B (multiple endocrine neoplasia type 2B), and familial medullary thyroid carcinoma. The variations in the RET gene play an important role in the MEN2 syndromes. In this work, we have developed a RET oligonucleotide microarray of 67 oligonucleotides to quickly detect RET mutations in MEN2 syndromes. The predominant RET mutations are missense mutations and are restricted to nine codons (codons 609, 611, 618, 620, 630, 634, 768, 804, and 918) in MEN2 syndromes. Missense mutations at codons 609, 611, 618, 620, and 634 have been identified in 98% of MEN2A families and in 85% of familial medullary thyroid carcinoma families. More than 95% of MEN2B patients also had a predominant mutation type at codon 918 (Met-->Thr). RET oligonucleotide microarray can detect RET missense mutations at these nine codons. Theoretically, a total of 55 missense mutation types can occur at eight codons (codons 609, 611, 618, 620, 630, 634, 768, and 804). RET oligonucleotide microarray is designed to detect all of these 55 missense mutation types at these eight codons and one predominant type at codon 918. Fifty-six oligonucleotides were designed for the 56 mutation types at nine codons, and 11 oligonucleotides were designed for the wild types and positive controls. We found RET mutations in all eight of the Korean MEN2A families (a total of 75 members; 27 affected members, 19 gene carriers, and 29 unaffected members) using the developed RET oligonucleotide microarray and an automatic sequencing. Because we found only five mutation types from eight MEN2A families, the international collaborations are required to see whether the RET oligonucleotide microarray may be used as a genetic diagnostic tool. Taken together, the RET oligonucleotide microarray can function as a fast and reliable genetic diagnostic device, which simplifies the process of detecting RET mutations.
2型多发性内分泌腺瘤病（MEN2）综合征以常染色体显性方式遗传，具有高外显率。它有三个亚型，即MEN2A（2A型多发性内分泌腺瘤病）、MEN2B（2B型多发性内分泌腺瘤病）和家族性甲状腺髓样癌。RET基因的变异在MEN2综合征中起重要作用。在本研究中，我们开发了一种包含67个寡核苷酸的RET寡核苷酸微阵列，以快速检测MEN2综合征中的RET突变。MEN2综合征中主要的RET突变是错义突变，且局限于九个密码子（密码子609、611、618、620、630、634、768、804和918）。在98%的MEN2A家族和85%的家族性甲状腺髓样癌家族中已鉴定出密码子609、611、618、620和634处的错义突变。超过95%的MEN2B患者在密码子918处也有主要的突变类型（甲硫氨酸→苏氨酸）。RET寡核苷酸微阵列可以检测这九个密码子处的RET错义突变。理论上，八个密码子（密码子609、611、618、620、630、634、768和804）总共可出现55种错义突变类型。RET寡核苷酸微阵列旨在检测这八个密码子处的所有55种错义突变类型以及密码子918处的一种主要突变类型。针对九个密码子处的56种突变类型设计了56个寡核苷酸，针对野生型和阳性对照设计了11个寡核苷酸。我们使用开发的RET寡核苷酸微阵列和自动测序在所有八个韩国MEN2A家族（共75名成员；27名患病成员、19名基因携带者和29名未患病成员）中发现了RET突变。由于我们在八个MEN2A家族中仅发现了五种突变类型，因此需要开展国际合作以确定RET寡核苷酸微阵列是否可作为一种基因诊断工具。综上所述，RET寡核苷酸微阵列可作为一种快速且可靠的基因诊断设备，简化了检测RET突变的过程。
