上海交通大學(xué)覃文新研究組在《柳葉刀·腫瘤學(xué)》發(fā)表腫瘤轉(zhuǎn)化醫(yī)學(xué)最新成果
上海交通大學(xué)醫(yī)學(xué)院附屬仁濟(jì)醫(yī)院上海市腫瘤研究所“癌基因及相關(guān)基因國(guó)家重點(diǎn)實(shí)驗(yàn)室”覃文新研究組，6月26日在國(guó)際著名的醫(yī)學(xué)雜志《柳葉刀·腫瘤學(xué)》（Lancet Oncology）發(fā)表題為“血清DKK1蛋白作為肝癌診斷標(biāo)志物的大規(guī)模臨床多中心研究”的研究論文,，證明DKK1蛋白可作為腫瘤標(biāo)志物用于肝細(xì)胞癌的血清診斷,。這是我國(guó)科學(xué)家首次在國(guó)際一流的臨床腫瘤學(xué)和腫瘤轉(zhuǎn)化醫(yī)學(xué)雜志上發(fā)表具有原始創(chuàng)新和自主知識(shí)產(chǎn)權(quán)的腫瘤血清蛋白標(biāo)志物Ⅱ期試驗(yàn)研究論文。
主持這一研究的是“癌基因及相關(guān)基因國(guó)家重點(diǎn)實(shí)驗(yàn)室”覃文新研究員,。在國(guó)家重點(diǎn)實(shí)驗(yàn)室主任王紅陽院士,、楊勝利院士和顧健人院士的支持下，覃文新研究員及研究團(tuán)隊(duì)于2003年首次發(fā)現(xiàn)并證明分泌蛋白DKK1（Dickkopf-1）在人類多種腫瘤包括肝癌中特異高表達(dá),，并在人類多種腫瘤細(xì)胞的培養(yǎng)上清和肝癌患者血清中檢測(cè)到其分泌性高表達(dá),，可作為腫瘤血清蛋白標(biāo)志物用于肺癌、肝癌,、乳腺癌,、宮頸癌等惡性腫瘤的血清診斷，上述創(chuàng)新發(fā)現(xiàn)已于2005年和2006年申請(qǐng)了中國(guó)發(fā)明專利和國(guó)際專利并在國(guó)內(nèi)外公開,，隨后其研究結(jié)果得到了國(guó)內(nèi)外實(shí)驗(yàn)室的廣泛驗(yàn)證,。在此基礎(chǔ)上，2008年覃文新研究員率領(lǐng)研究團(tuán)隊(duì)設(shè)計(jì)并開展了腫瘤血清蛋白標(biāo)志物DKK1用于肝細(xì)胞癌血清診斷的大規(guī)模臨床多中心試驗(yàn)研究,。該臨床試驗(yàn)由我國(guó)重要的肝癌臨床中心和研究基地復(fù)旦大學(xué)附屬中山醫(yī)院,、第二軍醫(yī)大學(xué)東方肝膽外科醫(yī)院、蘇州大學(xué)附屬第一醫(yī)院和上海交通大學(xué)醫(yī)學(xué)院附屬仁濟(jì)醫(yī)院上海市腫瘤研究所四家單位共同完成,，研究結(jié)果以“快速通道”形式在《柳葉刀·腫瘤學(xué)》發(fā)表,。主要?jiǎng)?chuàng)新發(fā)現(xiàn)如下：作為腫瘤血清蛋白標(biāo)志物，DKK1蛋白對(duì)肝細(xì)胞癌總體診斷的敏感性可達(dá)69.1%,、特異性為90.6%,；特別是對(duì)早期肝細(xì)胞癌（BCLC 0+A）和小肝癌（單個(gè)小于2厘米）的診斷敏感性分別可達(dá)70.9%和58.5%,、特異性分別為90.5%和84.7%；同時(shí),，DKK1蛋白能夠彌補(bǔ)甲胎蛋白（AFP）對(duì)肝細(xì)胞癌診斷能力的不足,，對(duì)甲胎蛋白陰性（低于20納克/毫升）肝細(xì)胞癌的診斷敏感性為70.4%、特異性為90%,，并可從甲胎蛋白陽性（高于20納克/毫升）的慢性乙型肝炎及肝硬化等高?；颊咧需b別診斷肝細(xì)胞癌，鑒別診斷敏感性達(dá)69.1%,、特異性為84.7%,；DKK1蛋白與甲胎蛋白聯(lián)合應(yīng)用，可將肝細(xì)胞癌總體診斷率提高至88%,；此外,，手術(shù)后患者血中的DKK1濃度迅速下降，血清DKK1蛋白亦可作為肝癌療效監(jiān)測(cè)和預(yù)后判斷指標(biāo),。
世界衛(wèi)生組織（WHO）2008年的統(tǒng)計(jì)資料顯示：全球每年肝癌新發(fā)病例約75萬人,、死亡約70萬人。肝細(xì)胞癌是肝癌的主要類型,、約占肝癌的80%以上,，全球每年半數(shù)以上的新發(fā)和死亡病例發(fā)生在中國(guó)，是危害我國(guó)人民生命健康的重大疾病,。由于診斷方法的局限性,，大多數(shù)肝細(xì)胞癌患者就診時(shí)已屬中晚期，失去了最佳治療時(shí)機(jī),。因此,，發(fā)現(xiàn)新的肝細(xì)胞癌診斷標(biāo)志物特別是血清診斷標(biāo)志物，及時(shí)有效的早查,、早診,、早發(fā)現(xiàn)，是提高患者生存率,、降低死亡率的關(guān)鍵之一,。目前臨床常規(guī)診斷和篩查中廣泛使用的腫瘤血清標(biāo)志物基本為血清蛋白標(biāo)志物，主要原因在于腫瘤血清蛋白標(biāo)志物除具有非侵襲性特點(diǎn)外,，還具有所需血清樣本量少（通常少于100微升）,、無需對(duì)血樣進(jìn)行抽提純化等前處理、檢測(cè)方法簡(jiǎn)單易推廣,、可自動(dòng)化快速批量檢測(cè),、重現(xiàn)性高、費(fèi)用低等優(yōu)點(diǎn),。DKK1就屬于腫瘤血清蛋白標(biāo)志物,，具備廣闊的臨床應(yīng)用前景,。
論文通信作者覃文新研究員致力于腫瘤微環(huán)境、腫瘤轉(zhuǎn)移和腫瘤早期診斷研究,。從腫瘤微環(huán)境中發(fā)現(xiàn)分泌蛋白DKK1可作為腫瘤血清蛋白標(biāo)志物到臨床試驗(yàn),，整個(gè)研究工作已歷經(jīng)十年。按《柳葉刀·腫瘤學(xué)》雜志的發(fā)表要求,，論文對(duì)腫瘤血清蛋白標(biāo)志物DKK1研究工作的首創(chuàng)性,、專利申請(qǐng)和發(fā)現(xiàn)歷史進(jìn)行了系統(tǒng)回顧。此外,，論文還對(duì)具有臨床應(yīng)用價(jià)值的腫瘤血清蛋白標(biāo)志物所應(yīng)具備的標(biāo)準(zhǔn)進(jìn)行了定義和討論,。近日從美國(guó)國(guó)家專利商標(biāo)局傳來好消息，所申請(qǐng)的國(guó)際專利“DKK1在癌癥診斷中的應(yīng)用”已獲得美國(guó)專利授權(quán),，為腫瘤血清蛋白標(biāo)志物DKK1診斷產(chǎn)品占領(lǐng)國(guó)際市場(chǎng)提供了知識(shí)產(chǎn)權(quán)保障,。據(jù)論文第一作者、博士研究生沈秋瑾介紹,，現(xiàn)已初步研制出具有自主知識(shí)產(chǎn)權(quán)的血清DKK1蛋白檢測(cè)系統(tǒng)，進(jìn)一步的后續(xù)臨床試驗(yàn)在有關(guān)單位的通力協(xié)作下正在進(jìn)行中,。腫瘤血清蛋白標(biāo)志物DKK1的臨床轉(zhuǎn)化工作,，還需在國(guó)家有關(guān)部門、上海市政府和企業(yè)的共同推進(jìn)下,，進(jìn)行腫瘤血清蛋白標(biāo)志物DKK1的產(chǎn)業(yè)化,，盡早在臨床應(yīng)用，造福廣大癌癥患者,。
該研究得到國(guó)家科技重大專項(xiàng),、國(guó)家973計(jì)劃和國(guó)家自然科學(xué)基金等項(xiàng)目的共同資助。（生物谷Bioon.com）
doi:10.1016/S1470-2045(12)70233-4
Serum DKK1 as a protein biomarker for the diagnosis of hepatocellular carcinoma: a large-scale, multicentre study
Qiujin Shen MSc a ?, Prof Jia Fan MD b c ?, Xin-Rong Yang PhD b ?, Prof Yexiong Tan PhD d ?, Prof Weifeng Zhao MD e ?, Yang Xu MD b, Ning Wang PhD a, Yongdong Niu PhD a, Zheng Wu BSc a, Prof Jian Zhou MD b, Prof Shuang-Jian Qiu MD b, Ying-Hong Shi MD b, Bin Yu PhD a, Ning Tang MSc a, Wei Chu BSc a, Min Wang BSc d, Jinhua Wu MSc e, Prof Zhigang Zhang PhD a, Prof Shengli Yang PhD a, Jianren Gu MD a, Prof Hongyang Wang MD a d, Prof Wenxin Qin PhD a
Summary
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.
