血清DKK1蛋白对肝细胞癌诊断及预后价值的临床Ⅲ期纵向回顾性试验及转化研究

肝细胞癌是严重危害人类健康的恶性肿瘤之一，早期诊断能有效降低其死亡率。甲胎蛋白是应用最广泛的肝细胞癌血清蛋白标志物，但其检测灵敏度和特异性一直备受争议。本课题组前期研究发现分泌蛋白Dickkopf-1（DKK1）在肝细胞癌患者癌组织和血液中高表达，并通过大规模临床多中心Ⅱ期试验研究，发现血清蛋白DKK1可有效诊断肝细胞癌，尤其是早期肝细胞癌和甲胎蛋白阴性肝细胞癌。基于前期结果，本项目设计了肿瘤标志物临床Ⅲ期纵向回顾性研究，即从一随访16年（1996-2012）的队列中，收集已患肝癌患者自纳入队列至患癌之间每年的血样，以及自治疗至肝细胞癌复发或死亡之间每年的血样，检测血清DKK1蛋白浓度，评估其是否能早于现有诊断手段提示患癌风险，及其对肝细胞癌的预后价值。为使该基础研究成果转化为临床应用，本项目同时拟研发具有自主知识产权的DKK1蛋白酶联免疫检测试剂盒，为肝细胞癌诊断和预后提供新的有效工具。

Hepatocellular carcinoma (HCC) is one of the malignant cancers which threaten the human health. Early diagnosis is an effective approach to decrease the death rate. Alpha-fetoprotein (AFP) is the most widely used serum protein marker for HCC. However, its sensitivity and specificity are unsatisfied so far. We previously found a secreted protein, Dickkopf-1 (DKK1), was overexpressed in the cancerous tissues and sera of the patients with HCC. Using a large-scale multicenter phase Ⅱ study, we found that serum DKK1 had the capability to diagnose HCC, especially early-stage HCC and HCC with negative AFP (≤20 ng/mL). Based on our previous results, we design a phase Ⅲ retrospective longitudinal repository study to further evaluate the significance of DKK1 for the diagnosis and prognosis of HCC. In a cohort which the participants were followed up from 1996 to 2012 (16 years), blood samples of the HCC patients were collected from the year of enrollment to the year of tumor detection, and from the year of therapy to the year of tumor recurrence or death. Serum DKK1 concentrations will be measured by ELISA, in order to assess whether DKK1 can diagnose HCC before the clinical diagnosis and predict HCC recurrence and survival. Furthermore, for translating our basic research to clinical application, we also plan to develop an ELISA kit for DKK1 protein detection with our own intellectual property and it may provide a useful tool for the diagnosis and prognosis of HCC.