在肿瘤血清蛋白标志物DKK1用于肝细胞癌血清诊断的大规模临床多中心试验研究中，癌基因及相关基因国家重点实验室覃文新研究组证明该蛋白可作为肿瘤标志物用于肝细胞癌的血清诊断。今天，他们的研究成果经“快速通道”，在线发表在《柳叶刀—肿瘤学》上。
据介绍，这是我国科学家首次在国际一流临床肿瘤学和肿瘤转化医学杂志上发表具有原始创新和自主知识产权的肿瘤血清蛋白标志物Ⅱ期试验研究论文。
通讯作者覃文新介绍说：DKK1蛋白对肝细胞癌总体诊断的敏感性为69.1%，特异性为90.6%；特别是对早期肝细胞癌和小肝癌的诊断敏感性为70.9%和58.5%，特异性为90.5%和84.7%；同时，DKK1蛋白能够弥补甲胎蛋白对肝细胞癌诊断能力的不足，对甲胎蛋白阴性肝细胞癌的诊断敏感性为70.4%、特异性为90%，并可从甲胎蛋白阳性的慢性乙型肝炎及肝硬化等高危患者中鉴别诊断肝细胞癌，鉴别诊断敏感性达69.1%、特异性为84.7%；DKK1蛋白与甲胎蛋白联合应用，可将肝细胞癌总体诊断率提高至88%；手术后患者血中的DKK1浓度迅速下降，血清DKK1蛋白亦可作为肝癌疗效监测和预后判断指标。
从肿瘤微环境中发现分泌蛋白DKK1到临床试验，整个研究工作已历经10年。按《柳叶刀—肿瘤学》杂志的要求，论文对DKK1研究工作的首创性、专利申请和发现历史进行了系统回顾。此外，论文对能在临床应用的肿瘤血清蛋白标志物所应具备的标准进行了定义和讨论。
此外，研究人员所申请的国际专利“DKK1在癌症诊断中的应用”已于近日获美国专利授权。
论文第一作者、博士研究生沈秋瑾说：“现已初步研制出具有自主知识产权的血清DKK1蛋白检测系统，后续工作正在进行中。” 覃文新则表示，该标志物的临床转化工作需要在政府和企业的共同推进下进行，以便尽早在临床应用。
该临床试验由复旦大学附属中山医院、第二军医大学东方肝胆外科医院、苏州大学附属第一医院和上海交通大学医学院附属仁济医院上海市肿瘤研究所等共同完成。
Serum DKK1 as a protein biomarker for the diagnosis of hepatocellular carcinoma: a large-scale, multicentre study
Shen Q, Fan J, Yang XR, Tan Y, Zhao W, Xu Y, Wang N, Niu Y, Wu Z, Zhou J, Qiu SJ, Shi YH, Yu B, Tang N, Chu W, Wang M, Wu J, Zhang Z, Yang S, Gu J, Wang H, Qin W.
BACKGROUND:
Hepatocellular carcinoma (HCC) is prevalent worldwide and improvements in timely and effective diagnosis are needed. We assessed whether measurement of Dickkopf-1 (DKK1) in serum could improve diagnostic accuracy for HCC.
METHODS:
We analysed data for patients with HCC, chronic hepatitis B virus (HBV) infection, liver cirrhosis, and healthy controls, recruited from two Chinese centres between December, 2008, and July, 2009. A validation cohort matched for age and sex was recruited from another centre in China between July, 2010, and June, 2011. DKK1 was measured in serum by ELISA by independent researchers who had no access to patients' clinical information. We used receiver operating characteristics (ROC) to calculate diagnostic accuracy.
FINDINGS:
We assessed serum DKK1 in 831 participants: 424 with HCC, 98 with chronic HBV infection, 96 with cirrhosis, and 213 healthy controls. The validation cohort comprised 453 participants: 209 with HCC, 73 with chronic HBV infection, 72 with cirrhosis, and 99 healthy controls. Levels of DKK1 in serum were significantly higher in patients with HCC than in all controls. ROC curves showed the optimum diagnostic cutoff was 2•153 ng/mL (area under curve [AUC] 0•848 [95% CI 0•820-0•875], sensitivity 69•1%, and specificity 90•6% in the test cohort; 0•862 [0•825-0•899], 71•3%, and 87•2% in the validation cohort). Similar results were noted for early-stage HCC (0•865 [0•835-0•895], 70•9%, and 90•5% in the test cohort; 0•896 [0•846-0•947], 73•8%, and 87•2% in the validation cohort). Furthermore, DKK1 maintained diagnostic accuracy for patients with HCC who were α-fetoprotein (AFP) negative (0•841 [0•801-0•882], 70•4%, and 90•0% in the test cohort; 0•869 [0•815-0•923], 66•7%, and 87•2% in the validation cohort), including for patients with early-stage HCC (0•870 [0•829-0•911], 73•1%, and 90•0% in the test cohort; 0•893 [0•804-0•983], 72•2%, and 87•2% in the validation cohort), compared with all controls. Raised concentrations of DKK1 in serum could differentiate HCC from chronic HBV infection and cirrhosis (0•834 [0•798-0•871], 69•1%, and 84•7% in the test cohort; 0•873 [0•832-0•913], 71•3%, and 90•6% in the validation cohort). Moreover, measurement of DKK1 and AFP together improved diagnostic accuracy for HCC versus all controls compared with either test alone (0•889 [0•866-0•913], 73•3%, and 93•4% in the test cohort; 0•888 [0•856-0•920], 78•5%, and 87•2% in the validation cohort).
INTERPRETATION:
DKK1 could complement measurement of AFP in the diagnosis of HCC and improve identification of patients with AFP-negative HCC and distinguish HCC from non-malignant chronic liver diseases.
FUNDING:
National Key Basic Research Programme of China, National Key Sci-Tech Special Projects of Infectious Diseases, National Natural Science Foundation of China, Research Fund for the Doctoral Programme of Higher Education of China.
文献链接： Serum DKK1 as a protein biomarker for the diagnosis of hepatocellular carcinoma: a large-scale, multicentre study
