尿液DNA甲基化在上尿路尿路上皮癌无创诊断中的应用价值

Thevalueofurine DNA methylation in non-invasive diagnosisof upper urinary tract epithelial carcinoma

目的探讨尿液DNA甲基化技术用于上尿路尿路上皮癌(UTUC)无创诊断的性能和价值。方法回顾性分析2019年8月至2021年3月中山大学孙逸仙纪念医院和北京大学第一医院收治的107例UTUC患者(UTUC组)和121例上尿路良性疾病患者(对照组)的病例资料。UTUC组男64例(59.8%),女43例(40.2%);年龄(67±10)岁;输尿管癌50例(46.7%),肾盂癌42例(39.3%),肾盂癌合并输尿管癌15例(14.0%);非肌层浸润性UTUC45例(42.0%),肌层浸润性UTUC51例(47.7%),分期不明确11例(10.3%);肿瘤分级低级别14例(13.1%),高级别93例(86.9%)。对照组男81例(66.9%),女40例(33.1%);年龄(52±14)岁;泌尿系结石104例(86.0%),输尿管狭窄或息肉13例(10.7%),单纯肾积水4例(3.3%)。UTUC组60例(56.1%)和对照组11例(9.1%)行尿脱落细胞学检查。UTUC组72例(67.3%)和对照组9例(7.4%)行尿液原位荧光杂交法(FISH)检查。对两组的尿沉渣样本行基于ONECUT2和VIM基因甲基化的定量PCR检测技术,≥1个基因阳性判定为阳性,2个均为阴性判定为阴性。以病理或临床诊断为标准,评估尿脱落细胞学、FISH、尿液DNA甲基化检查的敏感性、特异性、准确性、阳性预测值、阴性预测值。结果尿液DNA甲基化检查的总体敏感性为84.1%,特异性为90.9%,准确性为87.7%,阳性预测值为89.1%,阴性预测值为86.6%;敏感性显著高于尿脱落细胞学(43.3%,P<0.05)和FISH(69.4%,P<0.05);准确性明显高于尿脱落细胞学(52.1%,P<0.05)和FISH(71.6%,P<0.05);特异性与尿脱落细胞学(100.0%)和FISH(88.9%)差异均无统计学意义(P>0.05)。亚组分析结果显示,尿液DNA甲基化检查对不同部位肿瘤的诊断敏感性分别为输尿管癌74.0%(37/50例),肾盂癌90.5%(38/42例),肾盂癌合并输尿管癌100.0%(15/15例);对不同良性疾病的诊断特异性分别为泌尿系结石92.3%(96/104例),输尿管狭窄和息肉84.6%(11/13例),单纯肾积水75.0%(3/4例)。尿液DNA甲基化检查对低级别UTUC的诊断敏感性为64.3%,明显高于尿脱落细胞学的20.0%和FISH的36.4%(P<0.05);对高级别UTUC的诊断敏感性为87.1%,高于尿脱落细胞学的48.0%和FISH的75.4%(P<0.05);对非肌层浸润性UTUC的诊断敏感性为77.8%,明显高于尿脱落细胞学的29.2%和FISH的64.5%(P<0.05);对肌层浸润性UTUC的诊断敏感性为88.2%,优于尿脱落细胞学的53.6%和FISH的72.7%(P<0.05)。尿液DNA甲基化检查在中山大学孙逸仙纪念医院应用的敏感性和特异性分别为83.0%和92.0%,北京大学第一医院应用的敏感性和特异性分别为85.2%和85.7%。结论基于尿液ONECUT2和VIM基因甲基化技术用于UTUC诊断,具有较高的敏感性和准确性,优于尿脱落细胞学和FISH。

Objective To explore the performance and value of urine DNA methylation technology in non-invasive diagnosis of upper tract urothelial carcinoma(UTUC).Methods A retrospective analysis was conducted on the of 107 patients with UTUC(UTUC group)and 121 patients with benign upper urinary tract diseases(control group)admitted to Sun Yat sen Memorial Hospital and Peking University First Hospital from August 2019 to March 2021.There were 64 males(59.8%)and 43 females(40.2%)in the UTUC group,with the age of(67±10)years old.There were 50 cases(46.7%)of ureteral cancer,42 cases(39.3%)of renal pelvis cancer,and 15 cases(14.0%)of renal pelvis cancer combined with ureteral cancer.45 cases(42.0%)of non muscular infiltrative UTUC,51 cases(47.7%)of muscular infiltrative UTUC,and 11 cases(10.3%)of unclear stage.There were 14 cases(13.1%)with low tumor grade and 93 cases(86.9%)with high grade.There were 81 males(66.9%)and 40 females(33.1%)in the control group,with the age of(52±14)years old.There were 104 cases(86.0%)of urinary stones,13 cases(10.7%)of ureteral stenosis or polyps,and 4 cases(3.3%)of simple hydronephrosis.Sixty cases(56.1%)in the UTUC group and 11 cases(9.1%)in the control group underwent urine cytology examination.Seventy-two cases(67.3%)in the UTUC group and 9 cases(7.4%)in the control group underwent urine in situ fluorescence hybridization(FISH)examination.Quantitative PCR detection based on ONECUT2 and VIM gene methylation was performed on urine sediment samples from both groups.If≥1 gene was positive,it was determined as positive,and if both genes were negative,it was determined as negative.The sensitivity,specificity,accuracy,positive predictive value,and negative predictive value of urine cytology,FISH,and urine DNA methylation tests were evaluated based on pathological or clinical diagnosis criteria.Results The overall sensitivity of urine DNA methylation test was 84.1%,specificity was 90.9%,accuracy was 87.7%,positive predictive value was 89.1%,and negative predictive value was 86.6%,respectively.It's sensitivity and accuracy were significantly higher than that of urine cytology(43.3%,P<0.05)(52.1%,P<0.05)and FISH(69.4%,P<0.05)(71.6%,P<0.05).The accuracy was significantly higher than that of urine cytology(52.1%,P<0.05)and FISH(71.6%,P<0.05);There was no statistically significant difference in specificity between urine cytology(100.0%)and FISH(88.9%)(P>0.05).The results of subgroup analysis showed that the diagnostic sensitivity of urine DNA methylation test for tumors in different locations was 74.0%(37/50 cases)for ureteral cancer,90.5%(38/42 cases)for renal pelvis cancer,and 100.0%(15/15 cases)for renal pelvis cancer combined with ureteral cancer.The diagnostic specificity for different benign diseases was 92.3%(96/104 cases)for urinary stones,84.6%(11/13 cases)for ureteral stenosis and polyps,and 75.0%(3/4 cases)for simple hydronephrosis.The diagnostic sensitivity of urine DNA methylation test for low-grade UTUC was 64.3%,significantly higher than that of urine cytology(20.0%)and FISH(36.4%)(P<0.05).The diagnostic sensitivity for high-level UTUC was 87.1%,which was higher than that of urine cytology(48.0%)and FISH(75.4%)(P<0.05).The diagnostic sensitivity for non muscular invasive UTUC was 77.8%,which was significantly higher than that of urine cytology(29.2%)and FISH(64.5%)(P<0.05).The diagnostic sensitivity for myometrial infltrative UTUC was 88.2%,which was superior to urine cytology(53.6%)and FISH(72.7%)(P<0.05).The sensitivity and specificity of urine DNA methylation test in Sun Yat-sen Memorial Hospital of Sun Yat-sen University were 83.0%and 92.0%,respectively.It's sensitivity and specificity in Peking University First Hospital were 85.2%and 85.7%,respectively.Conclusions The urine ONECUT2 and VIM gene methylation assay have high sensitivity and accuracy in upper urinary tract urothelial carcinoma,which are superior to urine cytology as well as FISH.