多探针荧光原位杂交技术在膀胱尿路上皮癌诊断中的意义

摘要

目的：探讨多探针荧光原位杂交技术（fluorescence in situ hybridization，FISH）在膀胱尿路上皮癌诊断的可行性，并探讨其与肿瘤临床分期和病理分级的关系。方法收集经膀胱镜检查诊断为膀胱移行细胞癌35例患者，及10例非肿瘤病人，10例正常人群的新鲜尿液标本，应用 FISH检测膀胱尿路上皮癌患者尿液脱落细胞中的3、7、17、9号染色体异常，并进行尿细胞学检测（Urinary cytological examination，UCE）。统计学分析 FISH 和 UCE诊断膀胱癌的灵敏性和特异性，并分析 FISH检测染色体突变与膀胱肿瘤分期及分级的相关性。结果多探针联合 FISH 技术与UCE诊断膀胱癌的灵敏度分别为82．86％和37．14％（P＜0．05），特异度分别为95．0％和100％（P＞0．05）。3、7及17号染色体着丝粒特异性探针及9号染色体长臂2区1带（9p21）的 p16位点特异性探针联合应用诊断膀胱癌的灵敏度为82．86％，而单一探针诊断膀胱癌的灵敏度最高仅为65．71％。膀胱癌患者中发生3、7、17号染色体畸变及9号染色体 p16基因畸变均与不同病理分级（G1-G3）有关（P＜0．05）；3、7号染色体畸变与临床分期（T0-T4）有相关性（P＜0．05），而17号染色体及9号染色体 p16基因畸变与临床分期无相关性（P＞0．05）。结论多荧光探针 FISH诊断膀胱尿路上皮癌敏感性高、特异性强、无创，且与肿瘤分期和分级有一定相关性，临床应用价值高。%Objective To evaluate the fasibility of multiprobe fluorescence in situ hybridization (FISH)assay in diag-nosing urine bladder cancer,and to discuss the relationship of FISH and clinical stages and pathological grades.Methods Fresh urinary specimen was recruited from 35 bladder cancer patients who were diagnosed by biopsies,10 non-tumor patients,and 10 healthy individuals.Exfoliated cells from fresh urinary specimen was collected and analyzed by FISH u-sing centromeric probes of chromosome 3,7,17 and region probe of 9p21.Urinary cytological examination (UCE)was applied at the same time.The sensitivity and specificity of each method in the diagnosis of bladder cancer were deter-mined and various chromosomal aberrations and malignant degree of correlation by comparative analysis.Results The sensitivity of mutiprobe FISH and UCE in diagnosing bladder cancer were 82.86% and 37.14% respectively (P<0.05),the specificity of mutiprobe FISH and UCE were 95.0% and 100% respectively (P>0.05).The sensitivity of combine centromeric probes of chromosome 3,7,17 and region probe of 9p21 together was 82.86%,while the highest sensitivity using one of them was just 65.71%.The aberration frequency of chromosomes 3,7,17 and 9p21 were posi-tively associated with the pathological grades(G1-G3 )(P<0.05).Meanwhile,the abnormality of chromosomes 3 and 7 were positively associated with clinical stages (T0-T4 )(P<0.05),however,the aberration frequency of chromosomes 17 and 9p21 were proved not to be correlated with clinical stages of bladder cancer(P>0.05).Conclusion Mutiprobe FISH has high value for diagnosing bladder cancer because of its high sensitivity,accurate,non-invasive,and relation-ship with clinical stages and pathological grades.