WRN protein as a novel erythroblast immunohistochemical marker with applications for the diagnosis of Werner syndrome
WRN protein as a novel erythroblast immunohistochemical marker with applications for the diagnosis of Werner syndrome
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WRN 蛋白作为一种新型红细胞免疫组织化学标记物，在维尔纳综合征的诊断中具有应用价值
DOI：
10.1007/s00428-014-1703-6
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发表时间：
2014
期刊：
影响因子：
3.5
通讯作者：
Goto M.
中科院分区：
文献类型：
--
作者：
Sadahira Y;Sugihara T;Fujiwara H;Nishimura H;Suetsugu Y;Takeshita M;Okamura S;Goto M.
WRN基因突变的基因检测对于Werner综合征(WS)的诊断至关重要；然而，这些检测不能在临床环境中进行。几乎所有的WRN突变都会导致缺失C末端核定位信号的截短WRN蛋白的表达。我们通过石蜡包埋的骨髓切片评价了WRN蛋白免疫组织化学在诊断WS中的应用。使用一种针对WRN C末端的明确定义的商用多克隆抗体，我们发现在所有被测试的细胞类型中，骨髓红系前体细胞显示出最强的WRN核表达。对120例非WS血液病患者(年龄7天~90岁)的骨髓标本进行免疫组织化学分析，发现CD71阳性的早期和晚期红系前体细胞的细胞核均有WRN染色。WRN免疫染色阴性的红细胞仅见于2例，均诊断为WS：1例伴发骨髓增生异常综合征，1例红白血病伴TP53高表达。免疫细胞化学和Western印迹分析表明，WRN定位于4个阳性对照细胞系的细胞核中，而不定位于WS患者的5个阳性对照细胞系的细胞核中，WS患者的5个细胞系存在3种不同类型的WRN突变。因此，免疫组织化学检测骨髓石蜡切片中的WRN有助于WS的筛查，值得进一步的分子生物学证实。
Genetic testing for mutations in theWRNgene is critical for the diagnosis of Werner syndrome (WS); however, these tests cannot be performed in a clinical setting. Nearly all of theWRNmutations result in expression of truncated WRN proteins that are missing the C-terminal nuclear localization signal. We evaluated the use of WRN protein immunohistochemistry for diagnosing WS using paraffin-embedded bone marrow sections. Using a well-defined commercially available polyclonal antibody against the C terminus of WRN, we found that of all the cell types tested, bone marrow erythroid precursors showed the strongest nuclear expression of WRN. Immunohistochemical analysis of bone marrow samples from 120 patients with non-WS hematological disorders (age range, 7 days–90 years) revealed WRN staining of the nuclei of CD71-positive early and late erythroid precursors. Erythroblasts negative for WRN immunostaining were only observed in two patients, both of whom were diagnosed with WS: one with concomitant myelodysplastic syndrome and the other with erythroleukemia with overexpression of TP53. Western blot analysis and immunocytochemistry indicated WRN was localized in the nuclei of the four positive control cell lines from non-WS patients but not in the five cell lines from WS patients, who had three different types ofWRNmutations. Thus, immunohistochemical detection of WRN in erythroblasts from bone marrow paraffin sections could be useful in screening of WS cases and worthy of further molecular confirmation.