Polarity of prostate specific membrane antigen, prostate stem cell antigen, and prostate specific antigen in prostate tissue and in a cultured epithelial cell line
Polarity of prostate specific membrane antigen, prostate stem cell antigen, and prostate specific antigen in prostate tissue and in a cultured epithelial cell line
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DOI：
10.1002/pros.10203
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发表时间：
2003-04-01
期刊：
影响因子：
2.8
通讯作者：
Rajasekaran, AK
中科院分区：
文献类型：
--
作者：
Christiansen, JJ;Rajasekaran, SA;Rajasekaran, AK
背景资料。Madin-Darby犬肾(MDCK)细胞是一种永生化上皮细胞，已被广泛用作研究各种上皮细胞内分子转运、极化表达和蛋白质分泌的模型系统。为了确定MDCK细胞是否可以作为研究前列腺上皮细胞内分子事件的模型，我们在原位和MDCK细胞中检测了三种前列腺限制蛋白PSMA、PSCA和PSA的极化分布。方法采用免疫荧光、共聚焦显微镜、细胞表面生物素化、抗体内化和生化分析等方法，研究了三种前列腺限制蛋白在MDCK细胞中的表达和分泌。我们将这些表达模式与在前列腺上皮中观察到的结果进行了比较。结果：我们证明PSMA主要定位于前列腺上皮和转基因MDCK细胞的顶端质膜，而PSCA以非极化方式表达。我们还发现，PSA主要从转基因MDCK细胞的顶面分泌，这与体内观察一致。结论：MDCK和前列腺上皮细胞之间的定位模式相似，表明这些细胞类型中的极化分选机制是保守的。因此，MDCK细胞提供了一个有用的模型系统来研究这些蛋白质在前列腺内的靶向机制。(C)2003年Wiley-Liss，Inc.
BACKGROUND. Madin-Darby canine kidney (MDCK) cells are immortalized epithelial cells that have been used extensively as a model system to study intracellular molecular trafficking, polarized expression, and secretion of proteins in various epithelia. In order to determine if MDCK cells might serve as a model to study molecular events within prostate epithelial cells, we have evaluated the polarized distribution of three prostate restricted proteins, PSMA, PSCA, and PSA, in situ, and in MDCK cells.METHODS. Using immunofluorescence, confocal microscopy, cell surface biotinylation, antibody internalization, and biochemical assays we evaluated surface expression and secretion of three prostate restricted proteins expressed in MDCK cells. We compared these patterns of expression to results observed within prostatic epithelium.RESULTS. We demonstrate that PSMA is localized primarily to the apical plasma membrane in both the prostatic epithelium and transfected MDCK cells, whereas PSCA is expressed in a non-polarized fashion. We also show that PSA is secreted predominantly from the apical surface of transfected MDCK cells, consistent with in vivo observations.CONCLUSIONS. Similar patterns of localization among MDCK and prostatic epithelial cells suggest that the mechanisms of polarized sorting within these cell types are conserved. Thus, MDCK cells offer a useful model system to study mechanisms of targeting of these proteins within the prostate. (C) 2003 Wiley-Liss, Inc.