Uroplakins in the Lower Urinary Tract / 대한배뇨장애요실금학회지

The apical surface of mammalian urinary epithelium is covered by numerous scallop-shaped membrane plaques. This plaque consists of four different uroplakins (UPs) and integral membrane proteins. UPs, which are a member of the tetraspanin superfamily, are assembled into plaques that act as an exceptional barrier to water and toxic materials in urine. Within the plaques, the four UPs are organized into two heterodimers consisting of UP Ia/UP II and UP Ib/UP III in the endoplasmic reticulum. The two heterodimers bind to a heterotetramer, and then assemble into 16-nm particles in the Golgi apparatus. The aggregated UP complex ultimately covers almost all the mature fusiform vesicles in cytoplasm. These organelles migrate towards the apical urothelial cells, where they can fuse with the apical plasma membrane. As a result, the UPs are synthesized in large quantities only by terminally differentiated urothelial cells. For this reason, the UPs can be regarded as a major urothelial differentiation marker. In UP knockout (KO) mice, the incorporation of fully assembled UP plaques in cytoplasm into the apical surface is not functional. The mice with UP III-deficient urothelium show a significantly reduced number of UPs, whereas those with UP II-deficient urothelium have nearly undetectable levels of UPs. This finding strongly suggests that UP II ablation completely abolishes plaque formation. In addition, UP II KO mice contain abnormal epithelial polyps or complete epithelial occlusion in their ureters. UP IIIa KO mice are also associated with impairment of the urothelial permeability barrier and development of vesicoureteral reflux as well as a decrease in urothelial plaque size. In this review, I summarize recently published studies about UPs and attempt to explain the clinical significance of our laboratory results.

Expressions of Uroplakin gene family in interstitial cystitis / 中华泌尿外科杂志

Objective To detect gene expressions of Uroplakin (UP) family in interstitial cystitis (IC). Methods Gene expression of UP Ia, Ib, II, III, and III-δ4 was quantitatively measured in bladder biopsy samples from patients with IC (n=29) and control subjects (n=16) using real-time RT-PCR. Results UP III-delat4, a splicing variant of UP III, was significantly up-regulated to 4.80 in IC samples (P<0.001). When IC patients were divided into two groups, one with and one without ulcerative changes, the UP Ia, UP Ib, UP II, UP III and III-δ4 genes were significantly up-regulated to 4.99, 3.31, 2.75, 15.38, 11.48 in nonulcerative IC patients, but decreased to 0.04, 0.29, 0.09, 0.11, 0.34 in ulcerative IC patients (P<0.01). The up-regulation of UP III-δ4 was more prominent than that of UP III: 26.5-fold versus 5.6-fold compared to the median values of normal subjects. Conclusion Although the clinical implications of the over-expression of UP III and III-δ4 in non-ulcerative IC bladders remain to be clarified, from the diagnostic viewpoint, UP III-δ4 is a potential marker for identifying nonulcerative IC.

Uroplakin mRNA Expression in the Micro-dissected Mouse Prostate / 대한남성과학회지

PURPOSE: Prostatitis is a common condition with a significant effect on quality of life. Even though the etiology of chronic prostatitis remains unclear, certain bacterial infections may play a major role. In recent studies, E. coli, one important etiology of urinary tract infection, was found to mediate invasion into the bladder epithelium after binding uroplakin Ia in the apical membrane of the urinary bladder. Because E. coli is also an important pathogen for bacterial prostatitis, we investigated the uroplakin mRNA expression in micro-dissected mouse prostates. MATERIALS AND METHODS: We harvested the urinary bladder, ventral prostate, dorso-lateral prostate, and coagulating gland from 3 male imprinting control region (ICR) mice. The total RNA was extracted, cDNA was prepared, and finally the five target genes--uroplakin Ia, Ib, II, III, and beta-actin were amplified. We also examined the expressed sequence tags (EST) about above four uroplakin genes from mouse EST data. RESULTS: Uroplakin Ia, Ib, II, and III were expressed in the urinary bladder. However, only uroplakin Ia was definitively expressed in the ventral prostate. Uroplakin Ib and II were weakly expressed in the ventral, dorso-lateral, and coagulating prostate. Uroplakin III was not expressed in the prostate tissue. The mouse RNA transcripts in the EST data also showed similar results to uroplakin expression in the prostate. CONCLUSIONS: These results suggest that the mouse ventral prostate may be an adequate locus for acute or chronic bacterial prostatitis study. Further in-vitro bacteriologic studies of the ventral prostate will help reveal the mechanisms of chronic bacterial prostatitis.

Expressions of Uroplakins in the Mouse Urinary Bladder with Cyclophosphamide-Induced Cystitis

Even though uroplakins (UPs) are believed to serve a strong protective barrier against toxic materials, cyclophosphamide (CP) causes extensive cystitis. We investigated the expression of UPs in the urothelium in CP induced mouse cystitis. A total of 27 ICR female mice received a single intraperitoneal injection of 200 mg CP/kg. Nine CP-treated mice and 6 controls were sequentially killed at 12, 24, and 72 hr post injection. Extensive cystitis and an increased vesical weight were seen. These all peaked within 12 hr post injection and they tended to decrease thereafter. The level of all the UPs mRNA, the protein expressions of UP II and III on immunoblotting study, and the expression of UP III on immunolocalization study were maximally suppressed within 12 hr; this partially recovered at 24 hr, and this completely recovered at 72 hr post CP injection. In conclusion, CP reduced the expression of UPs. The reduction of the UPs mRNA and protein was time dependent, and this peaked within 12 hr after CP injection. However, the damage was rapidly repaired within 24 hr. This study demonstrates a dynamic process, an extensive reduction and rapid recovery, for the UPs expression of the mouse urinary bladder after CP injection.

Selection of Optimal Antisense Accessible Sites of Uroplakin Ⅱ mRNA for Bladder Urothelium / 华中科技大学学报（医学）（英德文版）

ysis, which had important reference values for further studying biological functions of UP Ⅱ gene and targeted therapeutic strategy for TCC of bladder.

Targeting gene therapy for bladder cancer using human UroplakinⅡ promoter / 中华泌尿外科杂志

Objective To investigate the feasibility of tissue-specific gene therapy for bladder cancer using human UroplakinⅡ(UPⅡ) promoter. Methods The recombinant adenoviruses,Ad-hUPⅡ-TNF carrying tumor necrosis factor (TNF-?) under control of the hUPⅡ,were generated.ELISA showed the production and secretion of TNF by bladder cancer cells infected with Ad-hUPⅡ-TNF.The level of TNF in urine was identified with ELISA. Results ELISA showed that production and secretion of TNF by bladder cancer cells infected with Ad-hUPⅡ-TNF was distinctly higher than by non-urothelium cells infected with Ad-hUPⅡ-TNF and MTT showed that proliferation of bladder cancer cells was obviously inhibited.Conditioned medium from bladder cancer cells apparently inhibited cells of L929 proliferation,compared with conditioned medium from non-bladder cells by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.Intravesical inoculation of Ad-hUPⅡ-TNF also caused decreased tumor growth in orthotopic human bladder cancer model.The sustained high level of TNF in urine could be identified with ELISA. Conclusions TNF driven by hUPⅡ promoter has effective active in the inhibition of bladder cancer growth both in vivo and in vitro.These will undoubtedly yield a new approach of therapy for bladder cancer.

Cloning of Human Uroplakin Ⅱ Gene from Chinese Transitional Cell Carcinoma of Bladder and Construction of Its Eukaryotic Expression Vector / 华中科技大学学报（医学）（英德文版）

To clone Uroplakin Ⅱ gene from Chinese transitional cell carcinoma (TCC) of bladder and construct its eukaryotic expression vector, the molecular cloning method was used to extract total RNA from a GⅢ/ T3N0M0 tissue sample of the bladder TCC patients. The primers were designed by Primer 5.0 software. Full length cDNA of Uroplakin Ⅱ gene was amplified by reverse transcription polymerase chain reaction (RT-PCR), assayed by nucleic acid sequencing and then inserted between Xba Ⅰ and HindⅢ restrictive sites of eukaryotic expression vector pcDNA3.0. The recombinant was assayed by restricted enzyme digestion. Under the induction of Lipofectamine 2000, the recombinant was transfected into Uroplakin Ⅱ negative bladder cancer cell line EJ. Cellular expression levels of Uroplakin Ⅱ were detected by RT-PCR. The nucleic acid sequencing results indicated that Chinese Uroplakin Ⅱ cDNA (555 bp) was successfully cloned. The BLAST analysis demonstrated that the cloned sequence is 100 % homologous with sequences reported overseas. The GenBank accession number AY455312 was also registered. The results of restricted enzyme digestion indicated that eukaryotic vector pcDNA-UP Ⅱ for Uroplakin Ⅱ was successfully constructed.After being transferred with pcDNA-UPⅡ for 72 h, cellular Uroplakin Ⅱ mRNA levels were significantly improved (P＜0.01). It is concluded that human Uroplakin Ⅱ gene was successfully cloned from Chinese TCC tissues, which provided a basis for further exploration of the roles of Uroplakin Ⅱ gene in TCC biological behaviors and potential strategies for targeted biological therapy of TCC.

Uroplakin Ⅱ gene in transitional cell carcinoma of the bladder / 中华泌尿外科杂志

Objective To investigate the expression of Uroplakin Ⅱ (UPⅡ) gene together with other conventional parameters in the patients with transitional cell carcinoma(TCC). Methods Total RNAs extracted from solid tumor tissues and blood samples of patients with TCC were reverse transcribed and subjected to polymerase chain reaction amplification (RT PCR).The products of RT PCR were analyzed by agarose gel electrophoresis. The expressions in solid tissues of other kinds of tumors and blood samples of non neoplasm patients served as controls. Results UPⅡ gene were expressed in all the solid tumor tissues of TCC(100%) and in the blood samples of 5 patients of T 3～T 4 neogrowth(39%), 2 of the 5 cases had hematogenous metastasis which became nondetectable after chemotherapy; UPⅡ gene has not been detected in the blood samples of T 1～T 2 TCC patients, the non neoplasm patients and other kinds of solid tumor tissues. Conclusions UPⅡ is one of the specific markers of urothelium.It can be found in TCC patients of stage T 3～T 4 and in patients with metastasis. The detection of UPⅡ in the blood sample might serve as a marker for chemotherapeutic efficacy.