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Objective:To develop a novel real-time fluorescence PCR method using Taqman probe for convenient,fast,sensitive and specific detection of urinary CK20 and use in early diagnosis and follow-up of transitional cell carcinoma of the bladder(TCCB).Methods:CK20 gene from cultured T24 cells was amplified by conventional RT-PCR,the standard quantitative plasmid was constructed by T-Aclone method.Taqman probe and primers was designed according to the sequence of CK20 cloned gene,the real-time PCR method for determination of CK20 mRNA was established and evaluated.Results:The developed real-time PCR method showed high sensitivity(10 2copies /?l) and good specificity;the linear range was 102 ~109 copies /?l,the coefficient variation(CV) was 0.89 % in intra-assay and 2.45% in day to day.Conclusion:A novel real-time fluorescence quantitative PCR method for detection of CK20 was established.It provided a basis for developing new diagnostic kits using the Taqman probe for clinical application.
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OBJECTIVE: To analyse some epidemiological data of patients with transitional cell carcinoma of the bladder (TCC) and the predictive value of conventional risk factors. METHODS: From 1980 to 2002, 125 patients were treated at our hospital and selected retrospectively for this study. The series mean age was 63,6±11,3 years, being 77.6% males and 22.4% females. The proportions of G!, G2 and G3 TCC were: 48%, 35.2% and 16.8%. The T stage was: pTa-1 - 85 (68%) and pT2-4 - 40 (32%). Superficial tumors were treated by TUR alone or plus BCG (>G1), and the invasives ones by radical cystectomy. The vesical reccurence rate of superficial TCC was 48.2%. RESULTS: TCC grade showed a positive association with T stage (p 0.0001) and patient survival (p 0.0001), but not with local bladder reccurence of superficial tumors (p=0.72). The T stage also showed a strong correlation with patient survival (p 0.0001). CONCLUSION: The TCC is more common in male than in female with mean age of 63.2 years. The tumor grading and staging exhibited as strong predictive value for disease progression and patient survival.
OBJETIVO: Investigar alguns dados epidemiológicos do câncer vesical bem como fatores de risco. MÉTODOS: Foram selecionados 125 pacientes atendidos no período 1980-2002, com idade média de 63,6±11,3 anos, sendo 97 (77,6%) homens e 28 (22,4%) mulheres. As proporções de tumores G1, G2 e G3 foram: 48%, 35,2% e 16,8%. As proporções dos estádios da lesão primária foram: pTa-1 - 85 ( 68%) e pT2-4 - 40 ( 32%). Entre os pacientes com tumores superficiais houve recidiva em 41 (48,2%). RESULTADOS: O grau da lesão primária mostrou relação com o estádio T (p 0,0001) e sobrevida (p 0,0001), mas não com a taxa de recidiva vesical dos tumores superficiais (p=0,72). O estádio T também mostrou forte correlação com a sobrevida (p 0,0001). CONCLUSÕES: O CCT é mais comum em homens que mulheres e acomete pacientes com idade média de 63,9 anos. O grau e o estádio T mostram forte correlação com a progressão da doença e a sobrevida.
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OBJECTIVE: To investigate the immunoexpression of p53 protein as a risk factor in transitional cell carcinoma of the bladder (TCC). METHODS: We analyzed retrospectively 90 patients with TCC and mean age of 71 years: G1 - 45, G1 - 29, G3 - 16, pTa-1 - 62 and pT2-4 - 28. The superficial TCC were treated TUR plus intravesical BCG (>G1), and the invasives by radical cystectomy and urinary diversion. The mean time of followup was 55 months and 25 patients died of the disease. The rate or reccurence in superficial tumors was 55.5%. The p53 immunoexpression was determined in formalin fixed paraffin embedded tumors sections by the avidin-biotin-immunoperoxidase method (DO - 7, Dako). We considered p53 positive TCC when the labeling nuclear index was >10%. RESULTS: P53 immunoexpression exhibited a positive association with tumor grade and stage (p=0.01), but not with the size of primary lesion (p=0.25) or rate of reccurence of pTa-1 tumors (p=0.81). A strong correlation was seen with metastases (p=0.002) and survival (p=0.003). CONCLUSION: A positive reaction for p53 showed correlation with tumor grade, T stage, metastases rate and parient survival, but had no predictive value for local reccurence rate of superficial tumors.
OBJETIVO: Investigar a expressão imuno-histoquímica da p53 com fator de risco em carcinoma de células transicionais da bexiga (CCT). MÉTODOS: Foram estudados restrospectivamente 90 pacientes com CCT com idade média de 71 anos: G1 - 45, G2 - 29, G3 - 16, pTa-1 - 62 e pT2-4 - 28. Entre os pacientes com tumores não invasivos houve recidiva vesical em 35 casos (55,5%). Os tumores superficiais foram tratados por ressecção trans-uretral associados ao BCG (>G1), e os invasivos por cistectomia radical. O tempo médio de seguimento dos pacientes foi de 55 meses e 25 deles faleceram da doença. A expressão imuno-histoquímica foi estudada em peças preservadas em formol 10% e blocos de parafina pelo método da avidina-biotina-imunoperoxidase. Considerou-se p53 positivo o tumor com índice de marcação nuclear superior a 10%. RESULTADOS: A expressão da p53 mostrou associação com o grau do tumor e com o estádio da lesão primária (p=0,01), mas não com o tamanho do tumor vesical (p=0,25) ou com a taxa de recidiva dos tumores superficiais (p=0,81). Houve forte correlação entre o padrão de marcação da p53 com metástases (p=0,002) e com a sobrevida dos pacientes (p=0,003). CONCLUSÃO: A expressão da p53 mostrou valor preditivo para grau tumoral, estádio, incidência de metástases e sobrevida dos pacientes, mas não para recidiva vesical dos tumores superficiais.
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Objective To investigate the expressions of survivin and bcl-2 and their relationships in transitional cell carcinoma of the bladder.Methods The expressions of survivin and bcl-2 in 40 cases of transitional cell carcinoma of the bladder were detected with streptavidin-biotin peroxidase(SP) method,with 15 cases of normal bladder specimens as controls.Results Survivin was expressed in 25 of 40(58.149%) cases of bladder carcinoma tissues,and normal bladder specimens did not express survivin(P
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The expression of proliferating cell nuclear antigen (PCNA) was assessed immunohistochemically in 47 cases of transitional cell carcinoma of the bladder and 3 cases of normal bladder with anti- PCNA/cyclin monoclonal antibody, using routinely processed tissue sections without interferring with histopathological diagnosis. The PCNA expression rates were compared with Ash histologic grade and clinical stage. In bladder cancer, the PCNA expression rate ranged from 3.8% to 32.7 % (mean value 11.2 %). Bladder cancer with Ash grade IV showed the highest PCNA expression rate (mean value 15.8 % ) and cancer with Ash grade I showed the lowest PCNA expression rate (mean value 8.3%). There were statistically significant differences of PCNA expression rates according to Ash grades (P=0.02. Kruskal-Wellis test). When clinical stage was analyzed to assess the relationship to PCNA expression rate invasive bladder cancers were associated ith higher PCNA expression rate then superficial bladder cancer (mean value of stage A; 8.7 %, stage B and C; 16.5 %). and the difference was statistically significant (P=0.003. Kruskal-Wallis test). Also, there was positive linear relationship between PCNA expression rate and Ash grade with regression analysis (r=0.573, P<0.0001, Y=4.41X +0.79). These results suggest that PCNA is useful as a unclear antigenic marker of cellular proliferation and offers an opportunity for analyzing cell kinetics successfully in formalin-fixed, paraffin-embeded tissue sections of transitional cell carcinoma of the bladder. It will be merited as a simple and powerful method to detect transitional cell carcinoma of the bladder with high potential of invasion, metastasis and clinical progression.