Borkowska Edyta Marta, Traczyk-Borszyńska Magdalena, Kutwin Piotr, Pietrusiński Michał, Jabłonowski Zbigniew, Borowiec Maciej
Medical University of Lodz, Chair of Laboratoty and Clinical Genetics, Department of Clinical Genetics, Lodz 92-213, Poland.
Medical University of Lodz, Chair of Laboratoty and Clinical Genetics, Department of Clinical Genetics, Lodz 92-213, Poland.
Urol Oncol. 2019 Dec;37(12):907-915. doi: 10.1016/j.urolonc.2019.06.010. Epub 2019 Jul 31.
The purpose of our research was to determine the usefulness of different methods for detecting Y373C mutation of gene FGFR3.
138 primary bladder cancer patients (71cases G1 and 67 cases G2-G3) were included in the study. Tumor tissue and urine samples were collected and kept frozen until the isolation of DNA. Sanger sequencing was applied for detecting mutation in cancer and ddPCR was utilized for urine assessment.
ddPCR appears to be more effective and it identified FGFR3 mutation (Y373C) in urinary sediment in 20.3% of cases whereas Sanger sequencing did in 15.5%. Only in 8/39 (20.5%) cases the mutation was observed both in urine and tissue. In 12/39 (30.8%) cases (5G1 and 7 G2-G3) we did not detect any FGFR3 mutation in urine although it was confirmed by sequencing. We only found mutation in urine in 20/39 cases (15 G1, 5 G2-G3) (51.3%). The correlation between the presence of FGFR3 mutations and better survival was confirmed. The Log-Rank test indicates a significant difference in the likelihood of survival for patients with the FGFR3 mutation but without recurrence (Cox's F-test P = 0.17006; Log-Rank Test P = 0.00059).
ddPCR appeared to be more sensitive method for detection FGFR3 gene mutation particularly for detecting low levels of tumor DNA amongst a large excess of nontumor DNA. It is significant as the implementation of such markers into routine practice could be beneficial. The prospective study in larger cohort is needed.
我们研究的目的是确定检测FGFR3基因Y373C突变的不同方法的实用性。
共138例原发性膀胱癌患者（71例G1期和67例G2 - G3期）纳入本研究。收集肿瘤组织和尿液样本并冷冻保存，直至提取DNA。采用桑格测序法检测肿瘤中的突变，采用数字液滴聚合酶链反应（ddPCR）进行尿液评估。
ddPCR似乎更有效，它在20.3%的病例中检测到尿沉渣中的FGFR3突变（Y373C），而桑格测序法为15.5%。仅在8/39（20.5%）的病例中，在尿液和组织中均观察到该突变。在12/39（30.8%）的病例（5例G1期和7例G2 - G3期）中，尽管测序证实存在FGFR3突变，但我们在尿液中未检测到任何突变。我们仅在20/39例（15例G1期，5例G2 - G3期）（51.3%）中发现尿液中有突变。FGFR3突变的存在与更好的生存率之间的相关性得到证实。对数秩检验表明，FGFR3突变但无复发的患者生存可能性存在显著差异（考克斯F检验P = 0.17006；对数秩检验P = 0.00059）。
ddPCR似乎是检测FGFR3基因突变更敏感的方法，特别是用于在大量非肿瘤DNA中检测低水平的肿瘤DNA。这具有重要意义，因为将此类标志物应用于常规实践可能有益。需要在更大队列中进行前瞻性研究。
