title: Value of reverse transcription polymerase chain assay in peripheral blood of patients with urothelial cancer.
abstract: OBJECTIVE
  The nested reverse transcription polymerase chain reaction (RT-PCR) method was used to determine expression of uroplakin II (UP II) or cytokeratin 20 (CK-20) in cells separated from the peripheral blood of patients with urothelial cancer. We examine whether UP II or CK-20 expression can be used as a urothelial cancer marker for urothelial cancer in cells isolated from peripheral blood.
  
  
  METHODS
  Peripheral blood was taken from 20 healthy volunteers without a history of urothelial cancer, from 10 patients with a negative bladder biopsy for urothelial cancer and from 108 patients with urothelial cancer. Results of a nested RT-PCR assay were compared with pathological stage and recurrence.
  
  
  RESULTS
  None of the peripheral blood samples from the control subjects revealed a positive polymerase chain reaction result. Among 108 patients with transitional cell carcinoma of the bladder nested RT-PCR for UP II was positive in 25 (23%) versus 31 (29%) for CK-20 (p >0.05). Nested RT-PCR for UP II was positive in 5 (8%) patients with superficial stage disease (pTa and pT1) versus 8 (11%) positive for CK-20. Nested RT-PCR for UP II was positive in 15 (58%) patients with a stage of pT2 or advanced stages versus 17 (65%) positive for CK-20. Nested RT-PCR for UP II was positive in 13 (20%) and 10 (56%) patients with grades 2 and 3, respectively, versus 17 (27%) and 12 (67%) nested RT-PCR positive for CK-20. A significant difference in the Kaplan-Meier recurrence-free actuarial curve was noted among patients with superficial stage who were positive and negative on nested RT-PCR for UP II and CK-20 in peripheral blood, respectively, but not in the invasive stage. On multivariate analysis nested RT-PCR for UP II and CK-20 in peripheral blood were independent prognostic factors in patients with superficial stage disease but not with invasive stage disease. Lung and/or liver metastasis developed in 5 (80%) of 6 patients whose results after chemotherapy (consisting of cisplatin, doxorubicin hydrochloride, vinblastine sulfate and methotrexate) for nested RT-PCR for UP II and CK-20 remained positive.
  
  
  CONCLUSIONS
  These results seem to indicate that UP II and CK-20 mRNA in the blood may be useful tumor markers for predicting patient survival and the extent of urothelial cancer.
authors.affiliation: Department of Urology, Kyorin University School of Medicine, Mitaka, Tokyo, Japan. toke@kyorin-u.ac.jp
authors.forename: Takatsuga
authors.initials: T
authors.lastname: Okegawa
authors.forename: Manami
authors.initials: M
authors.lastname: Kinjo
authors.forename: Kikuo
authors.initials: K
authors.lastname: Nutahara
authors.forename: Eiji
authors.initials: E
authors.lastname: Higashihara
mesh_terms: D000328:Adult; D001773:Blood Cells / Q000737:chemistry; D002295:Carcinoma, Transitional Cell / Q000097:blood* / Q000473:pathology; D005260:Female; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis*; D053554:Keratin-20; D008297:Male; D008565:Membrane Proteins / Q000032:analysis*; D008875:Middle Aged; D009367:Neoplasm Staging; D011237:Predictive Value of Tests; D011379:Prognosis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D001749:Urinary Bladder Neoplasms / Q000097:blood* / Q000473:pathology; D060170:Uroplakin II
chemical_list: D007381:Intermediate Filament Proteins; C507499:KRT20 protein, human; D053554:Keratin-20; D008565:Membrane Proteins; C554658:UPK2 protein, human; D060170:Uroplakin II