| Background and Purpose: Bladder cancer is the fifth most common human malignant diseases, and in the male urogenital system tumors, both of the morbidity and mortality account for the first. The main features of it is multi-lesion and a high rate of recurrence, 70% of superficial transitional cell carcinoma will recur after treatment, so patients with bladder cancer need to monitor recurrence. At present, the diagnosis and follow-up of bladder cancer patients mainly rely on the cystoscopy and urine cytology, but because of theirs invasion and low sensitivity , they are unsatisfactory. In recent years, more reports of fluorescence in situ hybridization in place of cystoscopy and cytology, with a non-invasion, high sensitivity and specificity, is a better means for the diagnosis and monitoring of bladder cancer recurrence.Compared with the cytology , FISH and cytology detection have a considerable specificity, but a higher sensitivity. It is reported that the sensitivity of urine cytology is about 63.8%, FISH is 80.4%. The specificity of cytology and FISH were 86.1% and 85.3%, with an insignificant difference. Moreover, the sensitivity of FISH can be increased with the tumor grade increased. So, compared to cytology, both in sensitivity and specificity, or for the lower grade tumors, FISH all have certain advantages.Fluorescence in situ hybridization technology is emerging in recent years, a molecular cytogenetic technique, which is a non-radioactive in situ hybridization developed on the basis of the radioactive in situ hybridization. The basic principle 5is to use known markers for the single-stranded nucleic acid probe, in accordance with the principle of base pair complementarity, With the combination of the unknown heterosexual single-stranded nucleic acid to form the double-stranded nucleic acid which can be detected by fluorescence microscopy . Technology itself is a relatively simple operation, have a good reproducibility, stability, and very effective sensitivity and specificity. Therefore, FISH has become a common means of detection widely used in prenatal and postnatal diagnosis of genetic diseases and hematologic malignancies, solid tumor and so on .Bladder cancer are characteristic with the variations of chromosome number and structure . common chromosomal abnormalities occur in the chromosome 3,7,17 and part of chromosome 9 deletions. Other such as the chromosome 1,4,8,10,11,13,14,20 and X, Y, may also occur the variations of chromosome number and structure .Currently,in Europe countries, the FISH technique with non-invasive, non-suffering has become one of the most important and effective method of the clinical detection for diagnosing bladder cancer or monitoring recurrence,. This experiment we had done aimed at cell morphology and FISH comparative study of technology in the Chinese population, in order to explore in the Chinese population the relationship between the chromosome 3, 7, 17 and p16 with bladder cancers' occurrence and development, establish the characteristics of China's population of bladder cancer molecular cytogenetic data , and provide a theoretical basis for the early diagnosis of urinary exfoliated cells of bladder cancer and monitoring the recurrence of bladder cancer.Method and subjects : According to existing research results, we select the four chromosomal genetic loci ,that is the chromosome 3,7,17 and p16 which related with occurrence and development of the bladder cancer. At the same time,all the subjects were divided into two groups, the experimental group and the control group. The experimental group, including the primary and recurrent bladder transitional cell carcinoma, and suspicion of urinary malignant tumors total 32 cases, all cases were designed in the blind, doing the FISH detection and cytology at the same time. The control group included 20 cases, doing the FISH detection, determinating of threshold.Results: CSP 3 and CSP 17 probe signal coverage throughout the centromere region, show the green fluorescence signal. CSP 7 probe signal coverage throughout the centromere region, show the orange fluorescence signal. And P16 probe signal coverage throughout the P16 gene, show the orange signal. In the fluorescence microscope, the nucleus are blue, for normal cells, there are two orange and two green signals. If not two, is unusual.The sensitivity of FISH detection is 96.8%, specificity is 50%. The sensitivity of cytology is 19.4%, specificity is 100%. In the experimental group , the chromosome aberration rate were significantly higher, the polyploid incidence of chromosome 3,7,9 p21 and 17 were 74.2%, 58.1%, 61.3%, 77.4%; the monomer incidence were 83.9 % , 83.9%, 96.8%, 67.7%; the homozygous deletion rate were 25.8%, 32.3%, 22.6%, 16.1%. In the experimental group, 31 cases of urothelial cell carcinoma, the chromosome aberrations of 3,7,9 p21and 17 had no significant correlation with the sex , age and single or multiple tumor of the patients (p> 0.05). The chromosome aberrations of 3,7and17 had no significant correlation in primary and recurrent tumor (p> 0.05); but the loss of 9p21 had (p = 0.041). Recurrent tumor had higher rate of loss of 9p21. The chromosome aberrations of 3 and 17 had no significant correlation between the stages of T 1 and T 2-4 (p> 0.05); but the chromosome polyploid of 7 and 9p21 had (p = 0.026 and 0.038). With the level increasing, the percentage of polyploid increased. The 3rd chromosome aberrations had no significant correlation between the grades of G 1 and G 2-3 (p> 0.05); but the chromosome aneuploidy of 7 had (p = 0.038). At the same time, the polyploidy of chromosome 7 and 17 had correlation between the grades of G 1 and G 2-3 (p = 0.025 and 0.047), while the polyploid of 9p21 had significant correlation (p = 0.0099). With the level increasing, the rate of aneuploidy increased. Conclusion: 1,FISH is a really effective means for detecting of chromosome variation . 2,Bladder carcinoma of urinary exfoliated cells exist a higher mutation rate of chromosome 3 , 7 , 9 p21 and 17 . 3,The sensitivity of FISH detection was significantly higher than the cytology , then the specificity is relatively poorer . 4,9 p21 aneuploidy was probably related to the recurrence,grades and stages of bladder cancer , chromosome 7 aneuploidy was probably related to the grades and stages of bladder cancer , and chromosome 17 probably only related to the grades . |
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