Molecular characterization of the loss of p75(NTR) expression in human prostate tumor cells.
Molecular characterization of the loss of p75(NTR) expression in human prostate tumor cells.
复制标题
人前列腺肿瘤细胞中 p75(NTR) 表达缺失的分子特征。
DOI：
10.1002/mc.1038
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发表时间：
2001
影响因子：
4.6
通讯作者：
Djakiew,D
中科院分区：
文献类型：
--
作者：
Krygier,S;Djakiew,D
低亲和力神经生长因子受体p75NTR是一种75 kDa的糖蛋白，属于肿瘤坏死因子受体超家族，参与诱导多种组织和细胞系的细胞凋亡。前列腺癌根治术组织切片免疫组织化学染色显示，p75NTR仅表达于上皮细胞。Western印迹和免疫组织化学分析也表明，在器官受限腺癌的恶性进展过程中，p75NTR在前列腺上皮细胞中的表达逐渐丧失，在来自转移的自然发生的前列腺癌细胞系DU-145、PC-3、LNCaP和TSU-PR1中完全丧失表达。将p75NTR表达重新导入TSU-PR1肿瘤细胞系，可以重新建立这些细胞经历p75NTR介导的凋亡的能力。目前尚不清楚这种表达缺失是由于p75NTR基因的部分或全部缺失还是其他因素。通过Southern杂交和聚合酶链式反应，我们发现p75NTR蛋白表达的缺失不是由于该基因的缺失或丢失。此外，通过逆转录-聚合酶链式反应、核糖核酸酶保护和染色质免疫沉淀实验，我们发现p75NTR基因在这些前列腺癌细胞系中发生了转录。最后，通过使用两个p75NTR的构建体瞬时转染，一个包含完整的2-kb 3‘非翻译区，另一个只包含几百个碱基的3’非翻译区，我们证明了3‘非翻译区可能在前列腺癌中p75NTR表达的缺失中起作用。©2001 Wiley-Liss公司
The low‐affinity nerve growth factor receptor p75NTRis a 75‐kDa glycoprotein that belongs to the tumor necrosis factor receptor superfamily and has been implicated in the induction of apoptosis in various tissues and cell lines. Immunohistochemistry on tissue sections from radical prostatectomies has shown that expression of p75NTRis limited to the epithelial cells. Western blot and immunohistochemical analyses have also shown a progressive loss of p75NTRexpression in prostate epithelial cells during the malignant progression of organ‐confined adenocarcinomas, with complete loss of expression in the naturally occurring prostate tumor cell lines DU‐145, PC‐3, LNCaP, and TSU‐pr1, which were derived from metastases. Reintroduction of p75NTRexpression into the TSU‐pr1 tumor cell line was shown to reestablish the ability of these cells to undergo p75NTR‐mediated apoptosis. It is not known whether this loss of expression is due to deletion of part or the entire p75NTRgene or to other factors. Through the use of southern blotting and polymerase chain reaction (PCR), we showed that loss of p75NTRprotein expression was not due to deletion or loss of the gene. Furthermore, through reverse transcription–PCR, RNase protection, and the chromatin immunoprecipitation assay, we showed that transcription of the p75NTRgene occurred in these prostate tumor cell lines. Finally, through transient transfection using two constructs of p75NTR, one containing the full 2‐kb 3′ untranslated region and one that contains only a few hundred bases of the 3′ untranslated region (UTR), we showed that the 3′ UTR may have a role in the loss of p75NTRexpression in prostate cancer. © 2001 Wiley‐Liss, Inc.