Department of Pathology, Research Institute and Hospital, National Cancer Center, Goyang, Korea.
Colorectal Cancer Branch, Research Institute and Hospital, National Cancer Center, Goyang, Korea.
Cancer Res Treat. 2016 Oct;48(4):1229-1242. doi: 10.4143/crt.2015.500. Epub 2016 Mar 30.
We developed a new method of detecting circulating tumor cells (CTCs) in liver cancer patients by constructing cell blocks from peripheral blood cells, including CTCs, followed by multiple immunohistochemical analysis.
Cell blockswere constructed from the nucleated cell pellets of peripheral blood afterremoval of red blood cells. The blood cell blocks were obtained from 29 patients with liver cancer, and from healthy donor blood spikedwith seven cell lines. The cell blocks and corresponding tumor tissues were immunostained with antibodies to seven markers: cytokeratin (CK), epithelial cell adhesion molecule (EpCAM), epithelial membrane antigen (EMA), CK18, α-fetoprotein (AFP), Glypican 3, and HepPar1.
The average recovery rate of spiked SW620 cells from blood cell blocks was 91%. CTCs were detected in 14 out of 29 patients (48.3%); 11/23 hepatocellular carcinomas (HCC), 1/2 cholangiocarcinomas (CC), 1/1 combined HCC-CC, and 1/3 metastatic cancers. CTCs from 14 patients were positive for EpCAM (57.1%), EMA (42.9%), AFP (21.4%), CK18 (14.3%), Gypican3 and CK (7.1%, each), and HepPar1 (0%). Patients with HCC expressed EpCAM, EMA, CK18, and AFP in tissue and/or CTCs, whereas CK, HepPar1, and Glypican3 were expressed only in tissue. Only EMA was significantly associated with the expressions in CTC and tissue. CTC detection was associated with higher T stage and portal vein invasion in HCC patients.
This cell block method allows cytologic detection and multiple immunohistochemical analysis of CTCs. Our results show that tissue biomarkers of HCC may not be useful for the detection of CTC. EpCAM could be a candidate marker for CTCs in patients with HCC.
我们通过构建包含循环肿瘤细胞（CTC）的外周血细胞细胞块，并进行多次免疫组织化学分析，开发了一种新的肝癌患者 CTC 检测方法。
去除红细胞后，从外周血有核细胞沉淀中构建细胞块。从 29 名肝癌患者和健康供体血液中加入七种细胞系的血液细胞块中获得血液细胞块。用七种标志物的抗体对细胞块和相应的肿瘤组织进行免疫染色：细胞角蛋白（CK）、上皮细胞黏附分子（EpCAM）、上皮膜抗原（EMA）、CK18、α-胎蛋白（AFP）、Glypican3 和 HepPar1。
从血细胞块中回收的 SW620 细胞的平均回收率为 91%。在 29 名患者中有 14 名（48.3%）检测到 CTC；11/23 例肝细胞癌（HCC）、1/2 胆管癌（CC）、1/1 合并 HCC-CC 和 1/3 转移性癌症。14 名患者的 CTC 对 EpCAM（57.1%）、EMA（42.9%）、AFP（21.4%）、CK18（14.3%）、Gypican3 和 CK（7.1%，各）、HepPar1（0%）呈阳性。HCC 患者在组织和/或 CTC 中表达 EpCAM、EMA、CK18 和 AFP，而 CK、HepPar1 和 Glypican3 仅在组织中表达。仅 EMA 与 CTC 和组织中的表达显著相关。在 HCC 患者中，CTC 检测与较高的 T 分期和门静脉侵犯相关。
这种细胞块方法允许对 CTC 进行细胞学检测和多次免疫组织化学分析。我们的结果表明，HCC 的组织生物标志物可能对 CTC 的检测没有用。EpCAM 可能是 HCC 患者 CTC 的候选标志物。
