IF 3 3区 医学 Q1 MEDICINE, GENERAL & INTERNAL Diagnostics Pub Date : 2025-03-03 DOI:10.3390/diagnostics15050607
Patrick T Gomella, Joon Yau Leong, Leonard G Gomella, Vivek S Tomar, Hector Teran, Edouard J Trabulsi, Madhukar L Thakur
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引用次数: 0

摘要

背景:诊断前列腺癌(PCa)的标准方法包括血清前列腺特异性抗原(PSA)检测、数字直肠检查(DRE)和图像引导下的靶向活检。鉴于这些技术的侵入性、潜在不良反应和成本,人们开始研究替代方法,特别是血清和尿液检测。本文介绍的工作旨在进一步验证一种用于 PCa 检测的新型非侵入性光学技术,该技术以前列腺癌恶性细胞(MC)上过度表达的 VPAC 基因组受体为目标,检测非 DRE 排出的尿液。检测方法在图像引导下进行活检并经组织学证实患有局部 PCa、计划进行前列腺癌根治术的患者(62 人)在手术前提供一份非 DRE 排空尿液样本。尿液经细胞离心后,将细胞固定在玻璃载玻片上,与 0.5 μg TP4303(本实验室开发的受体特异性荧光团,对 VPAC 具有高亲和力)一起孵育,过量清洗并用 4,6-二脒基-2-苯基吲哚(DAPI)进行核染色处理。使用蔡司 AX10 观察显微镜(20×)分析每张载玻片上的细胞视野。然后计算细胞和 MC 的总数,并使用 Zeiss 软件测量每个 MC 周围的荧光强度。此外,还对从临床确定的良性前列腺增生症患者(97 人)处收集的非 DRE 排空尿液样本进行了类似分析。结果对 62 名患者的尿液样本进行了处理和分析。按格里森等级(GG)分组的平均 PSA 水平分别为:GG1(10 人)6.5 ± 4.1 纳克/毫升,GG2(31 人)7.2 ± 3.8 纳克/毫升,GG3(13 人)13.2 ± 14.6 纳克/毫升,GG4(2 人)6.2 ± 2.2 纳克/毫升,GG5(6 人)50.2 ± 104.9 纳克/毫升。与 PSA 一样,GG5 型前列腺癌患者排出的尿液中 MC 的脱落率(66.7 ± 27.7)和荧光强度(35.8 ± 5.7)也最高。GG1至GG5的所有前列腺癌患者在排出的尿液中均有MC脱落,且MC脱落率和荧光强度不断增加,这与前列腺癌GG的增加有关。该检测方法对良性前列腺增生症的特异性为 89.6%(95% CI:81.9-94.9%),PPV 为 0.0%,NPV 为 100%(95.9% CI:95.9-100%)。结论这些数据表明(i)可以通过靶向 VPAC 受体检测非 DRE 排出的尿液中脱落的 PCa MC;(ii)非 DRE 排出的尿液中脱落的 MC 数量会随着疾病的严重程度而增加;(iii)这种非 DRE 排出的尿液光学检测法为初步检测 PCa 提供了一种简单、无创、可靠的方法,其成本可能低于目前可用的活检前检测技术。
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Non-DRE Voided Urine Test to Diagnose Prostate Cancer: Updated Results.

Background: The standard diagnostic approach for prostate cancer (PCa) diagnosis consists of serum prostate-specific antigen (PSA) testing, digital rectal examination (DRE) and image-guided targeted biopsies. Given the invasive nature, potential adverse events and costs associated with these techniques, alternative approaches have been investigated, specifically with serum and urine assays. The work presented here is intended to further validate a novel noninvasive optical technique for PCa detection, targeting the VPAC genomic receptors that are overexpressed on prostate cancer's malignant cells (MC), in non-DRE voided urine. Methods: Patients (N = 62) who had image-guided biopsy and histologically confirmed localized PCa, and who were scheduled for radical prostatectomy, provided a non-DRE voided urine sample prior to surgery. Urine was cytocentrifuged and cells fixed on a glass slide, incubated with 0.5 μg TP4303 (a receptor-specific fluorophore developed in our laboratory with high affinity for VPAC), excess washed and treated with 4,6-diamidodino-2-phenylindole (DAPI) for nuclear staining. The field of cells on each slide was analyzed using a Zeiss AX10 Observer microscope (20×). The total number of cells and MC were then counted, and the florescent intensity around each MC was measured using Zeiss software. Additionally, non-DRE voided urine samples collected from clinically determined BPH patients (N = 97), were also analyzed similarly. Results: Urine samples from 62 patients were processed and analyzed. Mean PSA levels by Gleason grade (GG) group were 6.5 ± 4.1 ng/mL for GG1 (N = 10), 7.2 ± 3.8 for GG2 (N = 31), 13.2 ± 14.6 for GG3 (N = 13), 6.2 ± 2.2 for GG4 (N = 2) and 50.2 ± 104.9 for GG5 (N = 6). Like the PSA, % MC shed (66.7 ± 27.7) in voided urine and the fluorescent intensity (35.8 ± 5.7) were highest in patients with GG5 prostate cancer. All PCa patients in GG1 to GG5 shed MC in voided urine with increasing % of MC and increasing fluorescence intensity which correlated with the increasing GG for PCa. For BPH, the specificity for the assay was 89.6% (95% CI:81.9-94.9%), PPV was 0.0% and NPV was 100% (95.9% CI, 95.9-100%). Conclusions: These data indicate the following: (i) PCa MC shed in non-DRE voided urine can be detected by targeting VPAC receptors, (ii) MC are shed in non-DRE voided urine with increasing quantity, corresponding to the severity of the disease, and (iii) this non-DRE voided urine optical assay provides a simple, noninvasive, and reliable method for the preliminary detection of PCa with potentially a lower cost than the currently available pre-biopsy detection technologies.

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来源期刊
Diagnostics
Diagnostics Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry
CiteScore
4.70
自引率
8.30%
发文量
2699
审稿时长
19.64 days
期刊介绍: Diagnostics (ISSN 2075-4418) is an international scholarly open access journal on medical diagnostics. It publishes original research articles, reviews, communications and short notes on the research and development of medical diagnostics. There is no restriction on the length of the papers. Our aim is to encourage scientists to publish their experimental and theoretical research in as much detail as possible. Full experimental and/or methodological details must be provided for research articles.
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