Fuja Tannin J, Lin Fritz, Osann Kathryn E, Bryant Peter J
Developmental Biology Center and Department of Medicine, University of California-Irvine, Irvine, California, USA.
Cancer Res. 2004 Feb 1;64(3):942-51. doi: 10.1158/0008-5472.can-03-2100.
We report somatic mutations in three genes (CSNK1 epsilon, encoding the Ser/Thr kinase casein kinase I epsilon; DLG1, encoding a membrane-associated putative scaffolding protein; and EDD/hHYD, encoding a progestin induced putative ubiquitin-protein ligase) in mammary ductal carcinoma. These genes were suspected of playing a role in cancer because loss-of-function mutations in their Drosophila homologues cause excess tissue growth. Using DNA from 82 laser-microdissected tumor samples, followed by microsatellite analysis, denaturing HPLC and direct sequencing, we found multiple somatic point mutations in all three genes, and these mutations showed significant association with loss of heterozygosity of closely linked polymorphic microsatellite markers. For CSNK1 epsilon and DLG1, most of the mutations affected highly conserved residues, some were found repetitively in different patients, and no synonymous mutations were found, indicating that the observed mutations were selected in tumors and may be functionally significant. Immunohistochemical reactivity of each protein was reduced in poorly differentiated tumors, and there was a positive association between altered protein reactivity, loss of heterozygosity, and somatic mutations. There was a statistically significant association of hDlg staining with p53 and Ki67 reactivity, whereas CSK1 epsilon and EDD/hHYD staining levels were associated with progesterone receptor status. The results provide strong indications for a role of all three genes in mammary ductal carcinoma. They also justify additional studies of the functional significance of the changes, as well as a search for additional changes in these and other genes identified from studies on model systems.
我们报告了乳腺导管癌中三个基因(编码丝氨酸/苏氨酸激酶酪蛋白激酶Iε的CSNK1ε、编码膜相关假定支架蛋白的DLG1以及编码孕激素诱导假定泛素蛋白连接酶的EDD/hHYD)的体细胞突变。这些基因被怀疑在癌症中发挥作用,因为其果蝇同源物中的功能丧失突变会导致组织过度生长。我们使用来自82个经激光显微切割的肿瘤样本的DNA,随后进行微卫星分析、变性高效液相色谱和直接测序,发现这三个基因均存在多个体细胞点突变,并且这些突变与紧密连锁的多态性微卫星标记的杂合性缺失显著相关。对于CSNK1ε和DLG1,大多数突变影响高度保守的残基,一些在不同患者中重复出现,且未发现同义突变,这表明观察到的突变在肿瘤中被选择,可能具有功能意义。在低分化肿瘤中,每种蛋白质的免疫组化反应性降低,并且蛋白质反应性改变、杂合性缺失和体细胞突变之间存在正相关。hDlg染色与p53和Ki67反应性之间存在统计学上的显著关联,而CSK1ε和EDD/hHYD染色水平与孕激素受体状态相关。这些结果强烈表明这三个基因在乳腺导管癌中均发挥作用。它们还证明有必要进一步研究这些变化的功能意义,以及寻找从模型系统研究中确定的这些基因和其他基因的其他变化。