Nucleolin Staining May Aid in the Identification of Circulating Prostate Cancer Cells
Nucleolin Staining May Aid in the Identification of Circulating Prostate Cancer Cells
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DOI：
10.1016/j.clgc.2016.12.004
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发表时间：
2017-06-01
影响因子：
3.2
通讯作者：
Pienta, Kenneth J.
中科院分区：
文献类型：
--
作者：
Chalfin, Heather J.;Verdone, James E.;Pienta, Kenneth J.
在晚期前列腺癌中，需要生物标志物来监测治疗反应并确定预后。目前循环肿瘤细胞 (CTC) 的检测依赖于敏感性和特异性有限的上皮标记物。我们发现核仁素（一种与增殖细胞相关的蛋白质）的染色模式有助于前列腺癌 CTC 的分类。简介：循环肿瘤细胞 (CTC) 作为实体恶性肿瘤的循环生物标志物具有巨大的潜力。目前可用的 CTC 检测方法依赖于敏感性和特异性有限的上皮标记物。我们发现核仁素（增殖细胞中常见的核仁蛋白）的染色模式可将患有转移性前列腺癌的男性中的 CTC 与白细胞 (WBC) 分开。患者和方法：使用 AccuCyte CTC 系统（RareCyte，西雅图，华盛顿州）处理 3 名患有转移性前列腺癌的男性的全外周血。使用 4',6-二脒基-2-苯基吲哚 (DAPI)、抗细胞角蛋白、抗 CD45/CD66b/CD11b/CD14/CD34 和抗核仁素抗体对载玻片进行免疫染色，并使用 CyteFinder 系统进行检测。使用新型图像分析软件测量 DAPI 核仁素共定位和染色模式小波熵。结果：使用新型成像软件对总共 33,718 个 DAPI 阳性细胞进行了分析，其中根据既定的 AccuCyte 系统标准，其中 45 个（0.13%）是已知的 CTC。可分段 CTC 的核仁素染色模式显示比 WBC 具有更大的小波熵（中位小波熵分别为 6.86 x 10(7) 和 3.03 x 10(6)；P = 2.92 x 10(-22)；近似 z 统计量 = 9.63）。此外，CTC 的总核仁素染色大于 WBC（中位总像素强度，分别为 1.20 x 10(5) 和 2.55 x 10(4) 积分像素单位；P = 2.40 x 10(-21)；近似 z 统计量 = 9.41）。结论：与 WBC 相比，前列腺癌 CTC 表现出独特的核仁素表达和定位。这一发现有可能成为灵敏且特异的 CTC 检测方法的基础。 (C) 2016 Elsevier Inc. 保留所有权利。
In advanced prostate cancer, there is a need for biomarkers to monitor response to therapy and determine prognosis. Current tests for circulating tumor cells (CTCs) rely on epithelial markers with limited sensitivity and specificity. We showed that the staining pattern of nucleolin, a protein associated with proliferative cells, aids in the classification of prostate cancer CTCs.Introduction: Circulating tumor cells (CTCs) have great potential as circulating biomarkers for solid malignancies. Currently available assays for CTC detection rely on epithelial markers with somewhat limited sensitivity and specificity. We found that the staining pattern of nucleolin, a common nucleolar protein in proliferative cells, separates CTCs from white blood cells (WBCs) in men with metastatic prostate cancer. Patients and Methods: Whole peripheral blood from 3 men with metastatic prostate cancer was processed with the AccuCyte CTC system (RareCyte, Seattle, WA). Slides were immunostained with 4',6-diamidino-2-phenylindole (DAPI), antiepan-cytokeratin, anti-CD45/CD66b/CD11b/CD14/CD34, and anti-nucleolin antibodies and detected using the CyteFinder system. DAPI nucleolin colocalization and staining pattern wavelet entropy were measured with novel image analysis software. Results: A total of 33,718 DAPI-positive cells were analyzed with the novel imaging software, of which 45 (0.13%) were known CTCs based on the established AccuCyte system criteria. Nucleolin staining pattern for segmentable CTCs demonstrated greater wavelet entropy than that of WBCs (median wavelet entropy, 6.86 x 10(7) and 3.03 x 10(6), respectively; P = 2.92 x 10(-22); approximated z statistic = 9.63). Additionally, the total nucleolin staining of CTCs was greater than that of WBCs (median total pixel intensity, 1.20 x 10(5) and 2.55 x 10(4) integrated pixel units, respectively; P = 2.40 x 10(-21); approximated z statistic = 9.41). Conclusion: Prostate cancer CTCs displayed unique nucleolin expression and localization compared to WBCs. This finding has the potential to serve as the basis for a sensitive and specific CTC detection method. (C) 2016 Elsevier Inc. All rights reserved.