BETA(2)-MICROGLOBULIN GENE-MUTATIONS - A STUDY OF ESTABLISHED COLORECTAL CELL-LINES AND FRESH TUMORS
BETA(2)-MICROGLOBULIN GENE-MUTATIONS - A STUDY OF ESTABLISHED COLORECTAL CELL-LINES AND FRESH TUMORS
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DOI：
10.1073/pnas.91.11.4751
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发表时间：
1994-05-24
影响因子：
11.1
通讯作者：
BODMER, WF
中科院分区：
文献类型：
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作者：
BICKNELL, DC;ROWAN, A;BODMER, WF
使用单链构象多态性 (SSCP) 技术筛选了一系列 37 个已建立的结直肠细胞系、22 个新鲜肿瘤样本和 22 个正常 DNA 样本中的 β(2)-微球蛋白基因突变。外显子1（包括前导肽序列）和外显子2分别进行筛选。 37 个结直肠细胞系中的 6 个和 22 个新鲜肿瘤中的 1 个被证明含有突变，而在正常 DNA 样本中未检测到突变。这些突变的测序表明，前导肽序列中的 8 bp CT 重复特别可变，因为 3 个细胞系和一个新鲜肿瘤样本在该区域存在缺失。在相关细胞系 DLD-1 和 HCT-15 中，鉴定出两个相似的突变，即密码子 10 中的 C --> A 替换和内含子 1 剪接序列中的 G --> T 突变。在 ELISA 系统中使用一系列单克隆抗体检查了 β(2)-微球蛋白的表达。表达减少与β(2)-微球蛋白的一个等位基因的突变相关，而在一个品系对于一个突变是纯合子或对于两个突变是杂合子的情况下，可以看到表达丧失。
The technique of single-strand conformation polymorphism (SSCP) was used to screen a series of 37 established colorectal cell lines, 22 fresh tumor samples, and 22 normal DNA samples for mutations in the beta(2)-microglobulin gene. Exon 1 (including the leader peptide sequence) and exon 2 were screened separately. Six of 37 colorectal cell lines and 1 of 22 fresh tumors were shown to contain mutations, whereas no mutations were detected in the normal DNA samples. Sequencing of these mutations showed that an 8-bp CT repeat in the leader peptide sequence was particularly variable, since 3 of the cell lines and one fresh tumor sample have deletions in this region. In the related cell lines, DLD-1 and HCT-15, two similar mutations were identified, a C --> A substitution in codon 10 and a G --> T mutation in the splice sequence of intron 1. Expression of beta(2)-microglobulin was examined using a series of monoclonal antibodies in an ELISA system. Reduced expression correlated with a mutation in one allele of beta(2)-microglobulin, whereas loss of expression was seen in instances where a line was homozygous for a mutation or heterozygous for two mutations.