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N41 Expression levels of P53 and CDKN2a/ARF messenger RNA detected by Real-Time PCR in tumor tissue in bladder cancer-clinicopathological applications

      Introduction and Objectives

      Cancer of the urinary bladder is a common malignant disease in the western countries.The majority of patients presents with superficial tumors with a high recurrence frequency, a minor fraction of these patients experience disease progression to muscle invasive stage. The present study aimed to asses wether patients with bladder urothelial tumors can be more objectively stratified into lowor high-risk groups for recurrence or progression.

      Material and Methods

      43 lessions were determind to be superficial papillary tumors (pTa), werehas 37 tumors invaded the lamina propria (pT1). Tumor grade was noted low (G1) in 46 cases and high (G2–3) in 34 cases. Reverse transcriptase Real Time PCR was performed in triplicate using iCycler iQ5 System (Bio-Rad).The rate of accumulation of amplified DNA was measured by continous monitoring of SBER Green fluorescence for P53 and GAPDH (reference gene), and by monitoring the level of fluorescence of Taqman probe for CDKN2a/ARF.Melt-curve analysis was performed immediately following amplifications.Tumor samples were evaluated for P53 and CDKN2a/ARF mutations using SSCP and sequencing methods.

      Results

      A total 80 patients (69 males) and 4 controls were enrolled into the study. Mean patients age was 68 years.Increase in expression level for P53 were observed in 5/43 pTa (11.6%) and 20/37 >pT1 (54%), for CDKN2a/ARF in 10/43 pTa (23%) and 2/37 >pT1 (5.4%). After a median follow-up 20 months (ranged from 6 to 45) 23/69 (33%) patients developed tumor recurrence and 11 died (2-^ months after first cystoscopy). An abnormal expression level were observed in 8/15 pTa (53%) cases and in 2/8 pT1 (25%) cases.There were no correlations between expression and mutations status.

      Conclusions

      We present data on the clinical usefulness of expression analysis in bladder carcinoma. Our data confirm that expression analysis is a promissing tool for bladder cancer diagnosis and prognosis.