| Objective: We evaluate the fluorescence in situ hybridization(FISH) assay for the diagnosis of bladder cancer in patients with hematuria through comparing the sensitivities and specificities of FISH and cytology for the detection of urinary bladder cancer.Methods: A total of 100 patients with gross or microscopic hematuria with suspected transitional cell cancer of bladder,collected from urology department of the second hospital of Hebei Medical University from April 2008 to December 2008,were included in the study, including 24 with a history of urinary bladder cancer and 76 without a history of urinary bladder cancer. Also we collected 20 healthy controls. Fluorescence in situ hybridization (FISH) is an application of non-radioactive fluorescent material which rely on the principle of nucleic acid probe hybridization in the chromosomal to show the location of DNA sequence, this method is rapid, non-invasive, high sensitive, which can detect DNA sequence and its changes in the medium term and interphase cells. This technology is widely used in tumor biology,gene mapping,gene amplification and other fields.FISH can detect various types of cytogenetic alterations including chromosome duplication, amplification, deletion.The FISH probe used for this study include one combination: GLP P16 probe/CSP 17 probe, P16 probe called the dependent kinase inhibitors 2A of cell cycle protein; CSP 17 probe called 17 chromosome centromere.The probe P16 hybridizate to human chromosome 9 long arm (9q21), covering the whole P16 genes, the fluorescent signal is red. For comparison, the probe CSP 17 hybridizate to human 17 chromosome centromere (17p11.1-q11.1), covering the entire centromere area, the fluorescent signal is green. A total of 120 urine specimens from 100 cases of patients with gross or microscopic hematuria with suspected transitional cell cancer of bladder and 20 healthy controls of different ages were analyzed in this research, A mixture of fluorescent labled probes to the centromeres of chromosome 17 and band 9q21 was used to detect exfoliated epithelial urinary cells for chromosomal abnormalities. FISH analysis was performed on 20 healthy controls of different ages. From counting 100 cells to chromosome 9q21 and 17 of 20 healthy controls and doing statistics to the number of different types of abnormal cells, We definited the threshold of chromosomal abnormalities. Meanwhile, FISH analysis was also performed on 100 patients ,applying the same method to count the number of different types of abnormal cells, The chromosomal results of 100 patients were defined through comparing with the threshold. A positive result was defined when 9 and 17 chromosomal abnormalities exist simultaneously or amplification and deletion occur in the same chromosome. Also Urine specimens from 100 suspected patients with BTCC were analyzed by means of cytology.The specimens for the cytology should be taken in the same day with FISH, and sent three times constantly. We divided the cytological results into positive , negative and suspicious results, then attributed the suspicious results which were difficult to diagnosis for bladder cancer to the positive in calculation. According to the different stages of bladder cancer we counted the number of positive cases including cytology and FISH , also compared them for their sensitivity and specificity. Voided urine was sent to a central laboratory for each study before cystoscopy, Suspicious lesions on cystoscopy were biopsied or resected. A centrally reviewed histopathological interpretation was used to confirm cancer and assign grade and stage.Results: A total of 93 patients was diagnosed histologically with transitional cell cancer of bladder or upper urinary tract.The sensitivity of FISH for pTis/pTa ( 22 cases) , pT1(52 cases),pT2(14 cases) and pT3/pT4(5 cases) tumors were 54.55%,88.46% ,92.86% and 100%,respectively.The sensitivity of urine cytology for pTis/pTa ( 22 cases) , pT1(52 cases),pT2(14 cases) and pT3/pT4(5 cases) tumors were 22.72 %, 38.46 % ,78.57% and 100%, respectively. For pTis/pTa tumors (P=0.030), For pT1tumors (P=0.001), For pT2 tumors (P=0.596).Overall sensitivity for FISH was significantly higher than the corresponding value for urine cytology (81.72% vs44.09%, respectively,p = 0.001). Specificities for FISH and cytology were 85.71% and 100%,respectively ( p = 0.299).Conclusion: The fluorescence in situ hybridization (FISH) assay is significantly more sensitive than voided cytology for detecting bladder cancer in patients evaluated for gross or microscopic hematuria for all stages while maintaining a similar specificity.FISH can improve significantly the diagnosis of urinary bladder cancer and will be significant for screening of urinary bladder cancer.This study provides theoretical basis for the establishment of early diagnostic methods to bladder cancer for Chinese people, which has nontraumatic, high specificity, high sensitivity.
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