应用荧光原位杂交技术检测膀胱尿路上皮癌的研究

The use of fluorescence in situ hybridization in the detection of urothelial bladder cancer

目的比较荧光原位杂交技术(FISH)与尿液脱落细胞学在膀胱尿路上皮癌检测中的灵敏性与特异性,评估FISH在中国人群中筛查膀胱尿路上皮癌的作用。方法 2007年12月至2008年6月,收取20例健康志愿者的新鲜尿液,应用荧光标记的3号、7号、17号染色体着丝粒探针及p16位点探针,在尿液标本中的脱落细胞上进行FISH检测,建立正常人群的阈值。其后留取无膀胱癌病史的119例血尿患者的尿液标本,膀胱镜检查前同期行尿液脱落细胞学检查及FISH检测。对比尿液脱落细胞学检查及FISH检测结果,对FISH、尿液脱落细胞学检查检测膀胱尿路上皮癌的敏感性和特异性采用McNemar配对卡方检验进行统计学分析。结果 3号、7号和17号染色体非整倍性改变及p16位点异常的正常阈值分别为1.0%、2.0%、1.0%和2.0%。对血尿患者进行的FISH、尿液脱落细胞学检测膀胱尿路上皮癌总的敏感性分别为83.1%(54/65)和52.3%(34/65)(P<0.001)。对于pTa～1期、pT2～4期肿瘤,FISH检测的敏感性分别为76.1%(35/46)和100.0%(19/19),而尿液脱落细胞学检查的敏感性分别为43.5%(20/46)和73.7%(14/19);对于低级别尿路上皮癌,FISH和尿液脱落细胞学的敏感性分别为72.2%(13/18)和5.6%(1/18),而对于高级别尿路上皮癌的敏感性分别为87.2%(41/47)和70.2%(33/47)。FISH检测、脱落细胞学检查的特异性分别为92.6%(50/54)、94.4%(51/54)(P=1.0)。FISH检测膀胱尿路上皮癌的敏感性明显优于尿液脱落细胞学检查,而两者的特异性并无统计学意义。结论 FISH技术能明显提高膀胱尿路上皮癌的筛查率,尤其是早期和低级别病变,可以成为筛查膀胱尿路上皮癌的一种新的无创性检查方法。

Objective We determine the relative sensitivities and specificities of cytology and fluorescence in situ hybridization(FISH) for the detection of urothelial bladder cancer in order to evaluate the clinical utility of FISH in the screening of urothelial bladder cancer.Methods From December 2007 to June 2008,the fresh urine voided by 20 healthy volunteers was obtained.A mixture of fluorescently labeled Probes were targeted to the chromosomes 3,7and 17 and p16.FISH was performed on exfoliated urothelial cells in the urine to get the threshold value of healthy adult.Urine voided by 119 patient with hematuria was collected and examined by cytology and FISH simultaneously before cystoscopy.No history of bladder cancer is required.The McNemar test was used to determine the statistical significance between the yields of the different tests.Results The threshold value of chromosomes 3,7 and 17 and p16 were 1.0%,2.0%,1.0% and 2.0% respectively.FISH affords an overall sensitivity of 83.1%(54 /65),the figure being 76.1%(35/46) for pTa ～ 1,100%(19 /19) for pT2 ～ 4 tumors.The sensitivities of cytology were 43.5%(20 /46) for pTa ～ 1,73.7%(14 /19) for pT2 ～ 4 tumors,with an overall sensitivity of 52.3%(34 /65) .The sensitivitiesof the two methods for the low grade tumors were 72.2%(13 /18) and 5.6%(1 /18),and for high grade tumors were 87.2%(41 /47) and 70.2%(33 /47),respectively.The specificities of FISH and cytology were 92.6%(50 /54) and 94.4%(51 /54),respectively(P = 1.0) .Conclusions According to the results,the sensitivity of FISH for the detection of urothelial bladder cancer is superior to that of cytology,and the specificities of FISH and cytology are not significantly different.FISH can improve markedly the screening rate of urothelial bladder cancer,especially for the low stage and low grade tumors,and will be a new noninvasive screening method for urothelial bladder cancer.