Quantitative detection of cytokeratin 20 mRNA in urine samples as diagnostic tools for bladder cancer by real-time PCR

• Veröffentlicht in: Experimental Oncology
• Datum: 2009
• Hauptverfasser: Guo, B., Luo, C., Xun, C., Xie, J., Wu, X., Pu, J.
• Format: Artikel
• Sprache: Englisch
• Veröffentlicht: Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України 2009
• Schlagworte: Original contributions
• Online Zugang: https://nasplib.isofts.kiev.ua/handle/123456789/134932
• Назва журналу: Digital Library of Periodicals of National Academy of Sciences of Ukraine
• Zitieren: Quantitative detection of cytokeratin 20 mRNA in urine samples as diagnostic tools for bladder cancer by real-time PCR / B. Guo1, C. Luo, C. Xun, J. Xie, X. Wu, J. Pu // Experimental Oncology. — 2009. — Т. 31, № 1. — С. 43-47. — Бібліогр.: 21 назв. — англ.

Aim: To determine and compare cytokeratin 20 (CK20) mRNA expression in urine of patients with transitional cell carcinomas
 of bladder (TCCB), urological benign diseases, and healthy volunteers. Methods: Taqman probe was designed according to the
 sequence of CK20 cloned gene. The quantitative PCR reaction system was optimized and evaluated. The CK20 mRNA level was
 screened by real-time polymerase chain reaction (RT-PCR) in 95 urine samples and analyzed according to the following parameters:
 urinary cytology, nuclear matrix protein 22 (NMP22) expression, tumor stage and grade. Results: For 60 TCCB patients urinary
 cytology was positive in 28 (46.7%), control group had no false-positive results (specificity 100%). CK20 expression was positive in
 RT-PCR of 51 cases (85%) of TCCB, but control group was positive in 2 cases (specificity 94.3%) with a cutoff value of crossover
 point (CT) = 30. Two methods have significant variation in sensitivity (p < 0.001), NMP22 expression was positive in 47 cases
 (78.3%), but control group was positive in 9 cases (specificity 85%). In the simultaneous evaluation of CK20 and NMP22 mRNA
 expression, there were 54 positive cases (90%). CK20 mRNA values in TCCB group (mean 27712.57 copies/µl) were significantly
 higher than in non-cancer disease urological group (mean 74.45 copies/µl) and control group (mean 8.47 copies/µl) (p < 0.001,
 p < 0.001, respectively). CK20 mRNA values increased gradually with higher tumor grade and stage: G1 differs significantly from
 G2 (p = 0.016); Tis/Ta
 differs significantly from T1–2 (p = 0.022). Conclusion: Our results indicate that CK20 mRNA expression
 could be regarded as a potential marker for TCCB. We demonstrated correlation between CK20 expression and the clinicopathologic
 features of TCCB (tumor stage and grade); simultaneous use of CK20 and NMP22 markers will elevate the sensitivity of the
 method. CK20 RT-PCR is a sensitive, quantitative, rapid and specific method to detect free cancer cells in the urine, and could be
 recommended for be wide application in the diagnostics of TCCB and evaluation of therapeutic effect.