Liebert M, Hubbel A, Chung M, Wedemeyer G, Lomax M I, Hegeman A, Yuan T Y, Brozovich M, Wheelock M J, Grossman H B
Department of Urology, University of Texas, Houston 77030, USA.
Differentiation. 1997 Feb;61(3):177-85. doi: 10.1046/j.1432-0436.1997.6130177.x.
We have developed an in vitro urothelial differentiation model. In this model, differentiated urothelial cells assemble desmosomes and E-cadherin at cell-cell junctions and stratify and show antigenic and functional evidence for tight junctions. Using this urothelial differentiation model with the differential display reverse-transcriptase polymerase chain reaction (ddRT-PCR), we identified two independently isolated gene fragments that showed near identity with the reported sequence for a human cDNA clone named mal. Differential expression of mal mRNA during urothelial differentiation was confirmed by RT-PCR using two other sets of PCR primers. Furthermore, uncultured urothelial cells from tissues also express mal mRNA, as indicated by RT-PCR. Mal was originally identified in a subtracted cDNA library as a human T-cell differentiation-associated gene and was thought to be T-cell specific. Our results identify mal as a gene also expressed in urothelial cells during differentiation and demonstrate the power of ddRT-PCR for analysis of gene expression under these controlled conditions.
我们开发了一种体外尿路上皮分化模型。在该模型中，分化的尿路上皮细胞在细胞间连接处组装桥粒和E-钙黏蛋白，分层排列，并显示紧密连接的抗原性和功能证据。利用这种尿路上皮分化模型和差异显示逆转录-聚合酶链反应（ddRT-PCR），我们鉴定出两个独立分离的基因片段，它们与一个名为mal的人类cDNA克隆的报道序列几乎完全相同。使用另外两组PCR引物通过RT-PCR证实了尿路上皮分化过程中mal mRNA的差异表达。此外，RT-PCR表明，来自组织的未培养尿路上皮细胞也表达mal mRNA。Mal最初在一个消减cDNA文库中被鉴定为人类T细胞分化相关基因，被认为是T细胞特异性的。我们的结果表明mal是一个在尿路上皮细胞分化过程中也表达的基因，并证明了ddRT-PCR在这些可控条件下分析基因表达的能力。
