液滴式数字PCR检测尿路上皮癌患者尿cfDNA中<i>FGFR3</i>基因S249C突变的临床应用价值

癌变·畸变·突变 ›› 2023, Vol. 35 ›› Issue (3): 165-171.doi: 10.3969/j.issn.1004-616x.2023.03.001

• 论著 •

液滴式数字PCR检测尿路上皮癌患者尿cfDNA中FGFR3基因S249C突变的临床应用价值

祖丽胡马尔·艾孜木阿吉¹, 赵焕², 马晟¹, 高苒³, 王雅茹¹, 肖汀¹

1. 国家癌症中心/国家肿瘤临床医学研究中心/中国医学科学院北京协和医学院肿瘤医院, 分子肿瘤学国家重点实验室, 癌发生及预防分子机理北京市重点实验室, 北京 100021;
2. 国家癌症中心/国家肿瘤临床医学研究中心/中国医学科学院北京协和医学院肿瘤医院, 病理科, 北京 100021;
3. 中国医学科学院医学实验动物研究所, 北京 100021

收稿日期:2023-03-12 修回日期:2023-04-28 发布日期:2023-06-03
通讯作者: 肖汀
作者简介:祖丽胡马尔·艾孜木阿吉,E-mail:zulhumar0102@163.com。
基金资助:
中国癌症基金会北京希望马拉松专项基金(LC2020A31)

Clinical value in using droplet digital PCR to detect urinary cfDNA FGFR3 S249C mutation in patients with urothelial carcinoma

ZULIHUMAER Aizimuaji¹, ZHAO Huan², MA Sheng¹, GAO Ran³, WANG Yaru¹, XIAO Ting¹

1. State Key Laboratory of Molecular Oncology, Beijing Key Laboratory for Carcinogenesis and Cancer Prevention, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021;
2. Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021;
3. Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China

Received:2023-03-12 Revised:2023-04-28 Published:2023-06-03

摘要/Abstract

摘要： 目的：通过非侵入性方式提取尿路上皮癌(UC)患者尿游离DNA(cfDNA)，使用液滴数字PCR(ddPCR)检测尿cfDNA中成纤维细胞生长因子受体3(FGFR3)基因S249C位点突变，辅助尿脱落细胞学检查提高UC的诊断效能。方法：收集27例UC患者尿液并提取尿cfDNA，采用ddPCR检测尿cfDNA中FGFR3 S249C突变情况。应用受试者工作特征曲线(ROC)评价尿cfDNA中FGFR3 S249C突变对于尿脱落细胞学诊断不明确UC患者的诊断效能。结果：UC患者尿cfDNA中FGFR3 S249C突变的检出率为25.9%(7/27)。按尿液细胞学巴黎报告系统(TPS)分类，FGFR3 S249C突变在高级别尿路上皮癌(HGUC)中检出率为18.8%(3/16)，可疑高级别尿路上皮癌(SHGUC)中为50%(4/8)，未见高级别尿路上皮癌(NHGUC)中为0(0/3)。FGFR3 S249C在非浸润性UC中的突变率为25.0%(1/4)，在浸润性UC中为31.6%(6/19)。在尿脱落细胞学诊断为SHGUC的样本中使用ddPCR检测出的FGFR3 S249C突变对于UC有50%(4/8)的检出率，且ROC曲线下面积为0.781，95% CI(0.407，1.000)，在UC诊断中起到良好的辅助作用。结论：使用ddPCR检测尿cfDNA中FGFR3 S249C突变作为一种非侵入性的检测方式可以辅助提高尿脱落细胞学诊断不明确样本的诊断效能。

关键词: 尿路上皮癌, 尿游离DNA, 液滴数字PCR, FGFR3 S249C突变, 尿脱落细胞学

Abstract: OBJECTIVE：The purpose of this study was to improve diagnostic efficacy of urothelial carcinoma (UC) by using the droplet digital PCR (ddPCR) to detect FGFR3 S249C (fibroblast growth factor receptor 3，FGFR3)mutation in cfDNAs which were extracted from urine of UC patients and in combination with urinary cytology. METHODS：Urine samples were collected from 27 UC patients and DNA samples were extracted. The FGFR3 S249C mutation in urinary cfDNA was detected by ddPCR. Receiver operating characteristic curves (ROC) were applied to evaluate the diagnostic efficacy of urinary cfDNA for patients with unclear urinary cytology but a clear pathological diagnosis of UC. RESULTS：The detection rate of the FGFR3 S249C mutation for UC was 25.9% (7/27). The detection rate of the mutation classified by TPS (The Paris system for reporting urinary cytology) was 18.8% (3/16) in HGUC (high-grade urothelial carcinoma)，50% (4/8) in SHGUC (suspicious for high-grade urothelial carcinoma)，and 0 (0/3) in NHGUC (negative for high-grade urothelial carcinoma). The mutation rate was 25.0% (1/4) in non-invasive UC and 31.6%(6/19)in invasive UC. The detection rate of positive mutation by ddPCR in SHGUC was 50.0% (4/8)，and the area under the ROC for the diagnosis of UC by the mutation was 0.781 with 95% CI (0.407，1.000)，which showed good accuracy for its diagnosis. CONCLUSION：Measurement of the FGFR3 S249C mutation in urinary cfDNA using ddPCR as a non-invasive assay could improve the diagnostic efficacy for UC with unclear urinary cytology diagnosis.

Key words: urothelial carcinoma, cfDNA, ddPCR, FGFR3 S249C mutation, urine cytology

中图分类号:

R730.4

祖丽胡马尔·艾孜木阿吉, 赵焕, 马晟, 高苒, 王雅茹, 肖汀. 液滴式数字PCR检测尿路上皮癌患者尿cfDNA中FGFR3基因S249C突变的临床应用价值[J]. 癌变·畸变·突变, 2023, 35(3): 165-171.

ZULIHUMAER Aizimuaji, ZHAO Huan, MA Sheng, GAO Ran, WANG Yaru, XIAO Ting. Clinical value in using droplet digital PCR to detect urinary cfDNA FGFR3 S249C mutation in patients with urothelial carcinoma[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2023, 35(3): 165-171.

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