荧光原位杂交(FISH)技术在尿路上皮肿瘤诊断中的应用研究

Study on diagnosis of urothelial carcinoma by fluorescence in situ hybridization(FISH)

目的应用荧光原位杂交(FISH)技术探索中国人群中3、7和17号染色体及9p21位点异常与尿路上皮肿瘤发生、发展的关系,评估其用于中国人群尿路上皮肿瘤诊断的可行性及临床应用前景。方法收集102例以血尿症状就诊,证实为尿路上皮肿瘤患者的新鲜晨尿,连续留取3d,将标本分别以FISH技术和尿脱落细胞学方法加以检测。收集20名正常成人新鲜晨尿,同法操作并计算正常成人对照组参考值。收集临床资料,明确肿瘤临床分期和病理分级。以正常成人对照组参考值判定患者FISH结果,比较其与尿脱落细胞学检测方法诊断的阳性检出率。统计分析3、7和17号染色体及9p21位点畸变率与性别、年龄、肿瘤初发、复发、单发、多发、临床分期及病理分级的相关性。结果①各染色体及区带均呈现较高的畸变发生率。多体畸变发生率由高到低为98.0%(7号)、97.1%(3号)、96.1%(17号)和83.3%(9p21位点);单体畸变发生率由高到低为95.1%(9p21位点)、68.6%(17号)、67.4%(7号)和66.7%(3号);缺失畸变则仅见于9p21位点(31.4%)。②尿脱落细胞学检查的肿瘤阳性检出率为64.7%,FISH则为98.0%,两者差异有统计学意义(χ2=30.25,P<0.01)。③各染色体畸变与患者的性别、年龄、肿瘤初发、复发、单发、多发及肿瘤临床分期均未见显著相关性。④3号染色体单体畸变与肿瘤病理分级间呈正相关(r=0.198,P=0.046),其余染色体不同类型的畸变情况均未显示出与不同病理分级间存在显著相关性。⑤3、7及17号染色体主要以多体畸变为主,而9p21位点主要以单体畸变为主(χ2=166.426,P<0.001),尚无证据表明各染色体与缺失畸变间的倍性关系。结论①FISH技术是通过尿液样本检测尿路上皮肿瘤染色体畸变的有效方法,可作为早期诊断尿路上皮肿瘤的重要辅助手段。②尿路上皮肿瘤在3、7和17号染色体及9p21位点同时存在多种畸变类型,且各畸变类型均具有较高的畸变率。③3号染色体的缺失或单体畸变可能对尿路上皮肿瘤的进展具有一定预测意义。

Objective To study the relations between aberrations of chromosome 3,7,17,9p21 and urothelial carcinoma,especially the bladder transitional cell carcinoma（BTCC）by the fluorescence in situ hybridization（FISH）.To evaluate the feasibility and the perspective of clinical applications of FISH in the diagnosis of urothelial carcinoma in Chinese group.Methods We collected the morning urine specimens of 102 patients who were diagnosed as urothelial carcinoma by biopsies or postoperative pathologic examinations,and then made tests on these specimens by FISH and uroexfoliative cytology respectively.The control group was set by detecting the morning urine specimens of 20 healthy persons using the technique of FISH in order to confirm the normal threshold value.The patients＇ positive results of FISH were defined according to the criteria made on the basis of the normal threshold value.Finally,we compared the positive detective rates between FISH and uroexfoliative cytology and analyzed whether the rates of abnormalities in chromosome 3,7,17,9p21 have significant associations with sex,ages,recurrence,multi-foci,clinical and pathologic tumor stages statistically.Results ①The polysomic aberration rates were 98.0%（7）,97.1%（3）,96.1%（17）,83.3%（9p21）;The monosomic aberration rates were 95.1%（9p21）,68.6%（17）,67.4%（7）,66.7%（3）;②The positive detective rates of FISH was 98.0%,while the uroexfoliative cytology was 64.7%.There was significant difference between these two techniques（χ2=30.25,P〈0.01）;③There were no correlation between abnormalities of chromosomes and sex,ages,recurrence,multi-foci and clinical stages significantly;④The polysomic aberration rates of chromosome 3 were associated with pathologic stage significantly（r=0.198,P=0.046）;⑤The main aberrative pattern of chromosome 3,7,17 was polysomy,chromosome 9p21 was monosomy（χ^2=166.426,P〈0.001）.Conclusions ①FISH is an effective method to detect chromosomal aberrations of urothelial carcinoma through the urine samples,which could also be ued as a good auxiliary technique in the diagnosis of urothelial carcinoma;②There exist both polysomic and monosomic aberrations in chromosome 3,7,17,9p21 and the high aberration frequency could be detected in each type of chromosomal abnormality;③The deletion and monosomic aberrations of chromosome 3 could be a indicative sign of tumor＇s progression.