Clinical and Experimental Pharmacology Group, Cancer Research UK Manchester Institute, University of Manchester, Manchester, UK.
Cancer Chemother Pharmacol. 2014 Aug;74(2):323-32. doi: 10.1007/s00280-014-2508-7. Epub 2014 Jun 13.
Our aims are to determine levels of circulating cellular and protein biomarkers in hepatocellular carcinoma (HCC) patients and to analyse any relationships with clinical parameters.
Fifty-four consenting patients were recruited. Circulating tumour cells (CTCs) were enumerated (by CellSearch) and characterised via filtration [by isolation by size of epithelial tumour cells (ISET)] with downstream immunohistochemistry (IHC). Glypican-3 (GPC3) expression in tumour biopsies and CTCs (by IHC) was compared, and levels of circulating caspase-cleaved and full-length cytokeratin 18 (CK18, measured using M30 and M65 ELISAs) were examined as a putative prognostic factor and marker of tumour burden.
CTCs were identified in 14 out of 50 (28%) patients by CellSearch and in 19 out of 19 (100%) patients by ISET. The presence of GPC3-positive CTCs by ISET was 100% concordant with the presence of GPC3-positive cells in the original tumour (n = 5). No statistically significant correlations were observed between CTC number and clinical characteristics, although trends were noted between CTC subtypes, Child-Pugh score and tumour node metastasis stage. Serum M30 and M65 levels (as continuous variables) significantly correlated with overall survival (OS) in a univariate analysis (p = 0.003 and p < 0.001, respectively); M65 levels remained statistically significant in a multivariate analysis (p = 0.029).
This is the first study to detect GPC3-positive CTCs in HCC, important for drug development with this target. The significant association of circulating CK18 with OS in HCC further exemplifies the utility of circulating biomarkers in cancer.
本研究旨在测定肝细胞癌（HCC）患者循环细胞和蛋白生物标志物的水平，并分析其与临床参数的关系。
共招募了 54 名同意参与的患者。通过 CellSearch 对循环肿瘤细胞（CTC）进行计数，并通过过滤[通过大小分离上皮肿瘤细胞（ISET）]对其进行特征分析，同时进行下游免疫组化（IHC）分析。比较肿瘤活检和 CTC 中的磷脂酰聚糖-3（GPC3）表达（通过 IHC），并检查循环半胱氨酸蛋白酶切割和全长细胞角蛋白 18（CK18 的水平，使用 M30 和 M65 ELISA 进行测量），作为一种潜在的预后因素和肿瘤负荷的标志物。
CellSearch 在 50 名患者中的 14 名（28%）患者中检测到 CTC，ISET 在 19 名患者中（100%）检测到 CTC。通过 ISET 检测到的 GPC3 阳性 CTC 与原始肿瘤中存在的 GPC3 阳性细胞（n = 5）完全一致。虽然在 CTC 亚型、Child-Pugh 评分和肿瘤淋巴结转移分期之间存在趋势，但未观察到 CTC 数量与临床特征之间存在统计学显著相关性。在单因素分析中，血清 M30 和 M65 水平（作为连续变量）与总生存期（OS）显著相关（p = 0.003 和 p < 0.001）；在多因素分析中，M65 水平仍然具有统计学意义（p = 0.029）。
这是首次在 HCC 中检测到 GPC3 阳性 CTC 的研究，这对于该靶点的药物开发很重要。循环 CK18 与 HCC 患者 OS 的显著相关性进一步证明了循环生物标志物在癌症中的应用价值。
