Hepatocellular carcinoma (HCC) is one of the most common types of cancer. Despite decades of research efforts, the search for novel biomarkers is still urgently needed for the diagnosis of HCC and the improvement of clinical outcomes. Previous studies of HCC clinical biomarkers have usually focused on serum and urine samples (e.g., serum Alpha-fetoprotein (AFP). However, cellular membrane proteins in lesion tissues are less used in HCC diagnosis. The abnormal expression of membrane glycoproteins in tumor lesions are considered as potential targets for tumor diagnosis and tumor therapies. Here, a lectin array has been employed to screen and identify abnormal glycopatterns and cellular membrane glycans in HCC lesion tissues compared with adjacent non-tumor tissues. We found that there was significantly less expression of Erythrina cristagalli (ECA) lectin binding (Galβ1-3/β1-4) glycans on the cellular membrane of HCC lesion tissues compared with those of adjacent non-tumor tissues. Immunohistochemistry analysis further showed that ECA-binding ability on the membrane proteins of HCC tissues progressively decreased in different tumor-node-metastasis (TNM) stages (stage I to stage III) as the malignancy of liver cancer increased. Receiver operating curve (ROC) analysis showed ECA-binding ability yielding a sensitivity of 85% and specificity of 75%, and a combination of ECA and AFP has better clinical diagnostic efficiency, yielding a sensitivity of 90% and specificity of 85%, than ECA or AFP assay alone. ECA pull-down followed by mass spectrometry further showed that there was significantly less expression of ECA binding membrane catalase (CAT) and prolyl 4-hydroxylase beta polypeptide (P4HB) in HCC tissues compared with the adjacent non-tumor tissues. The abnormally increased expression of total CAT and P4HB and decreased expression of galactosylated membrane CAT and P4HB in HCC cell lines were correlated with an HCC metastasis status. Our findings suggest that abnormal declined ECA-binding galatosylated membrane glycans and two galactosylated-CAT and P4HB glycoproteins in lesion tissues are potential biomarkers in the diagnosis and/or metastasis prediction for HCC.

肝细胞癌（HCC）是最常见的癌症类型之一。尽管进行了数十年的研究努力，但仍迫切需要寻找新的生物标志物来诊断 HCC 和改善临床结果。以往HCC临床生物标志物的研究通常集中在血清和尿液样本（如血清甲胎蛋白（AFP））。然而，病变组织中的细胞膜蛋白在HCC诊断中的应用较少。肿瘤病变中膜糖蛋白的异常表达是被认为是肿瘤诊断和肿瘤治疗的潜在靶点。在这里，凝集素阵列已被用于筛选和识别与邻近非肿瘤组织相比，HCC 病变组织中的异常糖模式和细胞膜聚糖。我们发现刺桐(ECA) 与相邻非肿瘤组织相比，HCC 病变组织细胞膜上的凝集素结合 (Galβ1-3/β1-4) 聚糖。免疫组织化学分析进一步表明，随着肝癌恶性程度的增加，ECA 对 HCC 组织膜蛋白的结合能力在不同的肿瘤淋巴结转移 (TNM) 阶段（I 期至 III 期）中逐渐降低。接受者操作曲线 (ROC) 分析显示 ECA 结合能力产生 85% 的敏感性和 75% 的特异性，ECA 和 AFP 的组合具有更好的临床诊断效率，产生 90% 的敏感性和 85% 的特异性，比单独进行 ECA 或 AFP 检测。ECA pull-down 随后质谱进一步表明，与邻近的非肿瘤组织相比，HCC 组织中 ECA 结合膜过氧化氢酶 (CAT) 和脯氨酰 4-羟化酶 β 多肽 (P4HB) 的表达明显减少。HCC 细胞系中总 CAT 和 P4HB 表达异常增加以及半乳糖基化膜 CAT 和 P4HB 表达减少与 HCC 转移状态相关。我们的研究结果表明，病变组织中异常下降的 ECA 结合半乳糖基化膜聚糖和两种半乳糖基化 CAT 和 P4HB 糖蛋白是 HCC 诊断和/或转移预测的潜在生物标志物。HCC 细胞系中总 CAT 和 P4HB 表达异常增加以及半乳糖基化膜 CAT 和 P4HB 表达减少与 HCC 转移状态相关。我们的研究结果表明，病变组织中异常下降的 ECA 结合半乳糖基化膜聚糖和两种半乳糖基化 CAT 和 P4HB 糖蛋白是 HCC 诊断和/或转移预测的潜在生物标志物。HCC 细胞系中总 CAT 和 P4HB 表达异常增加以及半乳糖基化膜 CAT 和 P4HB 表达减少与 HCC 转移状态相关。我们的研究结果表明，病变组织中异常下降的 ECA 结合半乳糖基化膜聚糖和两种半乳糖基化 CAT 和 P4HB 糖蛋白是 HCC 诊断和/或转移预测的潜在生物标志物。