Expression of CCN3 protein in human Wilms' tumors: immunohistochemical detection of CCN3 variants using domain-specific antibodies
Expression of CCN3 protein in human Wilms' tumors: immunohistochemical detection of CCN3 variants using domain-specific antibodies
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DOI：
10.1007/s00428-007-0523-3
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发表时间：
2008-01-01
期刊：
影响因子：
3.5
通讯作者：
Llombart-Bosch, Antonio
中科院分区：
文献类型：
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作者：
Subramaniam, Manish Mani;Lazar, Noureddine;Llombart-Bosch, Antonio
我们的目的是用抗K19M抗体和针对C端(CT)结构域和CCN3蛋白模块1、2、3和4的新型特异性抗体抗NH2、抗NH3、抗NH4和抗Nh5来检测8例Wilms瘤福尔马林固定石蜡包埋标本中截短的CCN3蛋白变体。在肾母细胞瘤中，除抗NH4抗体外，所有区域抗体在胚泡和原始小管中均可见胞核和胞浆染色。NH4仅在肿瘤细胞胞浆中检测到。正常胎肾除K19和NH5外，肾小管和肾小球中的抗体主要呈胞浆免疫反应，并有一定的核染色。我们的数据表明，在Wilms‘s肿瘤细胞中表达了一个截短的核CCN3变异体，缺乏凝血酶反应蛋白-1样结构域和细胞质全长CCN3蛋白。此外，正常胎儿肾脏主要表达全长蛋白，主要定位于细胞质。Wilms瘤细胞中截短的CCN3蛋白可能为其在这些肿瘤中的致瘤作用提供证据。与K19M的可变表达相比，NH5染色是一种更好的检测档案标本中CCN3蛋白CT域的方法。因此，区域特异性抗体是检测正常肾脏和肿瘤肾脏中CCN3蛋白变异的有价值的工具。
We aimed to detect truncated CCN3 protein variants in formalin-fixed paraffin-embedded samples of eight Wilms' tumors using anti-K19M and novel domain-specific antibodies, anti-NH2, anti-NH3, anti-NH4, and anti-NH5 raised against C-terminal (CT) domain and modules 1, 2, 3, and 4 of the CCN3 protein, respectively. In Wilms' tumors, all the domain antibodies except anti-NH4 exhibited both nuclear and cytoplasmic staining in blastema as well as primitive tubules. NH4 was detected only in the cytoplasm of tumor cells. Normal fetal kidneys revealed mainly cytoplasmic immunoreactivity for all antibodies in tubules and glomeruli, except for K19 and NH5, which showed some nuclear staining. Our data suggest expression of a truncated nuclear CCN3 variant lacking the thrombospondin type-1-like domain and cytoplasmic full-length CCN3 protein in Wilms' tumor cells. In addition, normal fetal kidneys express mainly full-length protein mostly localized to cytoplasm. Truncated CCN3 protein in Wilms' tumor cells may provide evidence for its tumorigenic role in these tumors. Uniform NH5 staining compared to variable expression of K19M indicates that using NH5 is a better approach for detecting the CT domain of CCN3 protein in archival specimens. Thus, the domain-specific antibodies represent valuable tools for detecting CCN3 protein variants in normal and neoplastic kidneys.