Expression of mal is associated with urothelial differentiation in vitro: identification by differential display reverse-transcriptase polymerase chain reaction.
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.
Liebert
M
M
Department of Urology, University of Texas, Houston 77030, USA.
Hubbel
A
A
Chung
M
M
Wedemeyer
G
G
Lomax
M I
MI
Hegeman
A
A
Yuan
T Y
TY
Brozovich
M
M
Wheelock
M J
MJ
Grossman
H B
HB
D001483:Base Sequence; D015820:Cadherins / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism; D051180:Desmoplakins; D006801:Humans; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D009185:Myelin Proteins; D062806:Myelin and Lymphocyte-Associated Proteolipid Proteins; D016133:Polymerase Chain Reaction / Q000379:methods*; D011510:Proteolipids / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics; D019459:Urothelium / Q000166:cytology / Q000502:physiology*
D015820:Cadherins; D003598:Cytoskeletal Proteins; D051180:Desmoplakins; C569026:MAL protein, human; D026901:Membrane Transport Proteins; D009185:Myelin Proteins; D062806:Myelin and Lymphocyte-Associated Proteolipid Proteins; D011510:Proteolipids; D012194:RNA-Directed DNA Polymerase