Zhang Lu, Hu Jin, Ali Zirakzadeh A, Rosvall Jesper, Hedlund Mats, Hu Ping Sheng, P A Wallin Robert, Sherif Amir, Winqvist Ola
Department of Medicine, Immunology and Allergy Unit, Karolinska University Hospital, SE-171 76, Stockholm, Sweden.
Department of Surgical and Perioperative Sciences, Urology and Andrology, Umeå University, Umeå, Sweden.
BMC Urol. 2018 Oct 5;18(1):86. doi: 10.1186/s12894-018-0399-3.
The tumor draining lymph node concept was first described in penile cancer for staging. Immunohistochemistry and histopathology evaluations are routinely used in clinical practice to examine lymph nodes for metastasis. However, these methods are time-consuming with low diagnostic accuracy and micro-metastases might be missed. In this study, we aim to evaluate detection of metastatic cells in draining lymph nodes by flow cytometry.
To assess the sensitivity of micro-metastasis detection by FACS (Fluorescence-activated cell sorting), HeLa cells were titrated into Peripheral blood mononuclear cells (PBMCs) and expression of pan-cytokeratin AE1/AE3 was analyzed. Single cell suspensions were separately prepared from 10 regional lymph nodes obtained from 5 patients with invasive penile cancer undergoing radical surgery and lymph node dissection. Lymph node dereived cells were examined for cell surface expression of EpCAM, E-cadherin and intracellular expression of pan-cytokeratin AE1/AE3 by FACS.
Ten lymph nodes from 5 penile cancer patients were investigated in a head-to-head comparison between FACS and pathology examination of sections. All metastatic lymph nodes verified by pathology examination were also identified by FACS. Two additional lymph nodes with micro-metastases were diagnosed by FACS only.
FACS analyses of pan-cytokeratin AE1/AE3 stained single cells from tumor draining lymph nodes can be used to detect micro-metastases in patients with penile cancer patients.
肿瘤引流淋巴结的概念最初是在阴茎癌分期中描述的。免疫组织化学和组织病理学评估在临床实践中常用于检查淋巴结是否转移。然而，这些方法耗时且诊断准确性低，可能会遗漏微转移。在本研究中，我们旨在评估通过流式细胞术检测引流淋巴结中的转移细胞。
为了评估荧光激活细胞分选术（FACS）检测微转移的敏感性，将HeLa细胞滴定到外周血单个核细胞（PBMC）中，并分析泛细胞角蛋白AE1/AE3的表达。从5例接受根治性手术和淋巴结清扫的浸润性阴茎癌患者获得的10个区域淋巴结中分别制备单细胞悬液。通过FACS检测淋巴结来源细胞的上皮细胞黏附分子（EpCAM）、E-钙黏蛋白的细胞表面表达以及泛细胞角蛋白AE1/AE3的细胞内表达。
在FACS与切片病理检查的直接比较中，对5例阴茎癌患者的10个淋巴结进行了研究。所有经病理检查证实的转移淋巴结也通过FACS得以识别。另外两个有微转移的淋巴结仅通过FACS诊断出来。
对肿瘤引流淋巴结中泛细胞角蛋白AE1/AE3染色的单细胞进行FACS分析可用于检测阴茎癌患者的微转移。
