Flow DNA Ploid Analysis And CYFRA21-1 In The Bladder Carcinoma And Its Clinical Significance

Bladder cancer is the most common malignancy of genitourinary system in China. It is essential to curative effect that early diagnosis and periodic detection after operation is performed. Cystoscopy is the gold standard for the detection of bladder tumors. But the clinical application is restricted by its invasive and some contraindications. Urine cytology is common in diagnosis of bladder cancer. It is sensitivity for bladder cancer was 20% to 70%. But it is influenced with the tumor malignancy grade and experience. In addition, some new tumor markers was used. Though the sensitivity of some tumor markers is better than urine cytology, its sensitivity and specificity is need to be increased. Therefore, it is important to find a useful clinical urine tumor marker with no invasion and high sensitivity and specificity .Though, there is a paper said that the analysis of urine exfoliated cell DNA can provide a basis in diagnosis of bladder carcinoma. The lack of a standardized methodology for quantifying DNA ploid by flow cytometry is hindering routine use of the marker in bladder cancer management. We validated a standardized flow cytometry protocol. The methodology of urinary CYFRA21-1 determination by electrochemiluminescent immunoassay is high sensitivity and specificity. The quantitative method that uses SYBR Green dye and 18SrRNA as internal control is suitable for clinical purpose. There were not reported that using quantitative real time PCR to detecting urinary CYFRA21-1 gene and the relationship between CYFRA21-1 and Flow DNA ploid. On the side, there are no papers reports about the relation between flow DNA ploid and CYFRA21-1. The other innovation is the combination of flow DNA ploid and CYFRA21-1 which is evaluated the diagnostic value of the non-invasive test for bladder transitional cell carcinoma of them.The result as follows:The levels of DI, SPF, PI, CV, G_O/G₁ and CYFRA21-1 in urine were significantly difference among the patients of bladder cancer, acute urethritis and normal control. The analysis of urine exfoliated cell DI, SPF, PI, CV, G_O/G₁ and CYFRA21 -1 can provide a basis in diagnosis of bladder carcinoma.There is no significantly difference in sex on urine exfoliated cell DI, SPF, PI, CV, G_O/G₁ and CYFRA21 -1 in bladder cancer patients. The levels of DI,SPF and CYFRA21-1 in the urine of bladder cancer increased with the development of the tumor grade and stage. Bladder carcinoma grade III and IV showed, in comparison to the grade I and II, significantly higher levels of DI. Bladder carcinoma stages Ta, T1, T2 and T3 is showed, in comparison to stages T4, significantly lower levels of PI and CYFRA21-1 contents. It is proved that urine exfoliated cell DNA ploid and CYFRA21-1 is associated with malignant transformation.An aneuploid content was observed in 20% of bladder cancer stage Ta and 62.5% in stage T1 to T4. The superficial bladder cancer (Ta ) showed, in comparison to the T1, T2, T3, T4 tumors, significantly lower numbers of aneuploid cells. It is showed that the rate of aneuploid in invasive bladder cancer significantly higher than superficial bladder cancer. It is proved that DNA aneuploidy is associated with progress of bladder cancer.There is no difference of the ratio of SPF>15% on pathological stages and grades of bladder cancer, but the ratio of SPF>15% is significantly higher than that of controls. So it is proved that SPF>15% can be one of bladder cancer marker.The mean CV of the G0-G1 peak exceeded 10% as standardized of bladder cancer. There is no difference of the mean CV of the G0-G1 peak on pathological stages and grades of bladder cancer, but the mean CV of the G0-G1 peak for bladder cancer samples is significantly higher than that of controls. So it is proved that the mean CV of the G0-G1 peak can be one of bladder cancer marker.There is no difference about DI, SPF, PI, CV, G_O/G₁ and CYFRA21-1 between the primary bladder cancer and recurrence. So the levels of DI, SPF, PI, CV, G_O/G1 and CYFRA21-1 were of no value in anticipating recrudescent bladder cancer.The cytometric analysis revealed that the DI and CV, G_O/G₁ were highly correlated with CYFRA21-1 in urine exfoliated cell of bladder cancer. The integration of the tests both flow DNA ploid and CYFRA21-1 into clinical practice takes evidently more than just the documentation of its sensitivity and specificity. It has important clinical practice value.Gene transcript level of CYFRA21-1 in Bladder carcinoma tissue and Para-cancerous tissue was detected by fluorescent quantitive real-time RT-PCR assay. CYFRA21-1 mRNA transcript level in cancerous tissues increased significantly compared with the para-cancerous tissues, it is concluded that there is a positive correlation between CYFRA21-1 and the incidence of bladder cancer. CYFRA21-1 can be a novel marker for bladder carcinoma.Gene transcript level of CYFRA21-1 in the urine of bladder cancer was detected by fluorescent quantitive real-time RT-PCR assay. It is concluded that there was significant positive correlation between the CYFRA21-1 protein expression and mRNA transcript level. CYFRA21-1 mRNA level in urine of bladder cancer correlated significantly with the malignant degree of tumors.The expressions of flow DNA ploid and CYFRA21-1 were significantly related to biological behavior of bladder cancer. The combined determination of urinary flow DNA ploid and CYFRA21-1 was more sensitivity and specificity, it maybe have higher value to diagnosis bladder cancer. It is a no invasion and good tumor maker which is valuable to monitor the curative effect.