血清ULBP-2、MIC-1聯(lián)合檢測(cè)診斷胰腺癌的價(jià)值

周郁芬 黃李雅 徐凌霄 張帆 國(guó)芳 姚瑋艷 袁耀宗

·論著·

血清ULBP-2、MIC-1聯(lián)合檢測(cè)診斷胰腺癌的價(jià)值

周郁芬 黃李雅 徐凌霄 張帆 國(guó)芳 姚瑋艷 袁耀宗

目的探討UL16結(jié)合蛋白2(ULBP-2)、巨噬細(xì)胞抑制因子-1(MIC-1)聯(lián)合檢測(cè)對(duì)胰腺癌診斷的價(jià)值。方法收集152例胰腺癌、20例胰腺癌前病變、91例慢性胰腺炎患者及96例健康對(duì)照者血清，應(yīng)用酶聯(lián)免疫吸附測(cè)定(ELISA)方法檢測(cè)ULBP-2、MIC-1水平，并和CA19-9水平進(jìn)行比較。采用受試者工作特征(ROC)曲線評(píng)估它們對(duì)胰腺癌的診斷價(jià)值。結(jié)果胰腺癌、胰腺癌前病變、慢性胰腺炎及健康對(duì)照者血清ULBP-2水平分別為(219.9±182.5)、(62.6±11.4)、(68.4±36.8)、(76.5±40.9)μg/L；MIC-1水平分別為(3521.3±3903.4)、(973.6±589.0)、(959.6±879.0)、(427.6±317.0)μg/L；CA19-9水平分別為(1448.8±3707.0)、(12.0±9.3)、(38.2±139.0)、(7.7±5.0)kU/L。胰腺癌患者均顯著高于癌前病變、慢性胰腺炎患者及健康對(duì)照者(χ2值分別為40.628、71.662、45.505，15.827、36.433、63.494，26.264、73.427、49.088，P值均<0.01)。ULBP-2、MIC-1、CA19-9診斷胰腺癌的曲線下面積(AUC)分別為0.909、0.818、0.864，三者聯(lián)合診斷的AUC為0.982，單指標(biāo)診斷以ULBP-2為佳，三者聯(lián)合診斷效能最高。對(duì)胰腺癌早期病變(胰腺癌前病變+胰腺癌ⅠA期病變)的診斷，ULBP-2、MIC-1、CA19-9的AUC分別為0.506、0.837、0.684，單指標(biāo)以MIC-1為佳，而MIC-1聯(lián)合CA19-9的診斷效能最高(AUC為0.897)。結(jié)論ULBP-2、MIC-1在胰腺癌患者血清中含量升高，二者聯(lián)合CA19-9檢測(cè)可提高對(duì)胰腺癌的診斷價(jià)值。

胰腺腫瘤； UL16結(jié)合蛋白2； 巨噬細(xì)胞抑制因子-1； 生物學(xué)標(biāo)記

雖然臨床上常用糖鏈抗原19-9(CA19-9)診斷和指導(dǎo)治療胰腺癌，但其敏感性和特異性均不高[1]。UL16結(jié)合蛋白2(UL-16 binding proteins 2,ULBP-2)屬于經(jīng)典MHC Ⅰ類相關(guān)分子，可與NK細(xì)胞激活受體NKG2D/DAP10結(jié)合，使NK細(xì)胞啟動(dòng)對(duì)靶細(xì)胞(腫瘤細(xì)胞、感染細(xì)胞)的殺傷作用，通過(guò)基質(zhì)金屬蛋白酶類(MMPs)的作用，成為血清中的游離分子[2]。巨噬細(xì)胞抑制因子-1(macrophage inhibitory cytokine-1，MIC-1)屬于轉(zhuǎn)化生長(zhǎng)因子β(TGF-β)超家族成員，在生理狀態(tài)下僅胎盤組織有高表達(dá)，但在炎癥、腫瘤組織中可以出現(xiàn)高表達(dá)[3]。ULBP-2、MIC-1與腫瘤的發(fā)生、發(fā)展密切相關(guān)。本研究旨在探討ULBP-2、MIC-1聯(lián)合檢測(cè)對(duì)胰腺癌的早期診斷價(jià)值。

材料與方法

一、病例資料

收集瑞金醫(yī)院消化科、外科2008年12月至2012年1月期間收治的152例經(jīng)手術(shù)病理、內(nèi)鏡超聲(EUS)細(xì)針穿刺活檢、ERCP胰管刷檢證實(shí)的胰腺癌患者。另選取胰腺癌前病變(手術(shù)病理確診為不典型增生者)20例、慢性胰腺炎患者91例、健康志愿者96例作為對(duì)照。收集患者一般情況、臨床表現(xiàn)及實(shí)驗(yàn)室檢查結(jié)果等資料。

二、方法

收集各組患者及健康對(duì)照者的血清，采用酶聯(lián)免疫吸附測(cè)定(ELISA)方法檢測(cè)血清ULBP-2、MIC-1及CA19-9的水平。ELISA試劑盒購(gòu)于R&D公司，按試劑盒說(shuō)明書操作[4-5]。

三、統(tǒng)計(jì)學(xué)處理

結(jié) 果

一、各組血清ULBP-2、MIC-1、CA19-9水平

胰腺癌、胰腺癌前病變、慢性胰腺炎患者及健康對(duì)照者血清ULBP-2水平分別為(219.9±182.5)、(62.6±11.4)、(68.4±36.8)、(76.5±40.9)μg/L；MIC-1水平分別為(3521.3±3903.4)、(973.6±589.0)、(959.6±879.0)、(427.6±317.0)μg/L；CA19-9水平分別為(1448.8±3707.0)、(12.0±9.3)、(38.2±139.0)、(7.7±5.0)kU/L。胰腺癌患者均較癌前病變、慢性胰腺炎患者及健康對(duì)照者顯著升高(χ2值分別為40.628、71.662、45.505，15.827、36.433、63.494，26.264、73.427、49.088，P值均<0.01)。胰腺癌前病變患者僅血清MIC-1水平較健康對(duì)照者顯著升高(χ2=12.274，P<0.01)。慢性胰腺炎患者的血清MCI-1及CA19-9水平均較健康對(duì)照者升高。

二、胰腺癌患者血清ULBP-2、MIC-1水平與腫瘤臨床病理特征間的關(guān)系

胰腺癌患者血清ULBP-2水平與患者的性別、年齡及腫瘤部位、T分期、TNM分期均無(wú)關(guān)；MIC-1水平與腫瘤的部位、T分期有關(guān)，而與患者的性別、年齡及腫瘤TNM分期無(wú)關(guān)(表1)。

三、ULBP-2、MIC-1、CA19-9診斷胰腺癌的效能

ULBP-2、MIC-1、CA19-9診斷胰腺癌的AUC分別為0.909、0.818、0.864(圖1)，ULBP-2>CA19-9>MIC-1。ULBP-2與MIC-1、ULBP-2與CA19-9、MIC-1與CA19-9聯(lián)合診斷的AUC分別為0.953、0.977、0.932。三者聯(lián)合診斷的AUC為0.982。以三者聯(lián)合診斷的效能最佳。

參數(shù)例數(shù)ULBP-2(μg/L)P值MIC-1(μg/L)P值年齡(歲) ≤6064152.8±188.60.2553197.0±3869.00.056 >6088229.0±178.43843.6±4052.0性別 男101211.3±177.80.5333545.7±3659.00.158 女51237.9±192.73470.4±4415.0腫瘤位置 胰頭99229.0±192.30.5034280.6±4485.70.025 胰體13178.9±135.61727.65±1017.3 胰尾36222.5±180.92278.5±2255.9 全胰腺4129.4±63.02711.7±1820.9T分期 T111325.8±392.10.8671648.3±1710.60.014 T222219.4±158.93942.6±3817.6 T363210.6±164.94124.1±4734.7 T455211.3±162.63499.9±3365.4TNM分期 ⅠA5105.0±43.40.3602227.4±292.80.166 ⅡB12206.3±143.51576.7±1820.8 ⅡA36222.5±175.14463.7±5047.2 ⅡB20242.4±161.33669.3±3980.1 Ⅲ40179.6±125.33190.2±3361.5 Ⅳ39264.1±257.23656.8±3673.7

ULBP-2、MIC-1、CA19-9診斷胰腺癌早期病變(胰腺癌前病變+胰腺癌ⅠA期病變)的AUC分別為0.506、0.837、0.684，MIC-1>CA19-9> ULBP-2。ULBP-2與MIC-1、ULBP-2與CA19-9、MIC-1與CA19-9聯(lián)合診斷的AUC分別為0.861、0.689、0.897。三者聯(lián)合診斷的AUC為0.894。以MIC-1和CA19-9聯(lián)合診斷的效能最佳(圖2)。

圖1ULBP-2、MIC-1、CA19-9診斷胰腺癌的ROC曲線圖2MIC-1、CA19-9聯(lián)合診斷胰腺癌早期病變的ROC曲線

討 論

Li等[6]報(bào)道，ULBP-2基因中有p53的反應(yīng)單元，這一單元的去甲基化使得p53與之結(jié)合，導(dǎo)致ULBP-2表達(dá)上調(diào)，從而激發(fā)抗腫瘤的固有免疫機(jī)制。臺(tái)灣學(xué)者Chang等[4]檢測(cè)了胰腺癌患者、健康對(duì)照者的血清ULBP-2水平，發(fā)現(xiàn)胰腺癌患者的血清ULBP-2在癌早期即明顯升高，對(duì)胰腺癌的診斷價(jià)值優(yōu)于CA19-9，若兩者聯(lián)合診斷可提高診斷的敏感性和特異性。但ULBP-2在其他消化系腫瘤中只有低水平表達(dá)，在其他系統(tǒng)腫瘤的血清中尚未發(fā)現(xiàn)表達(dá)增高。Yang等[7]報(bào)道，MIC-1是p53活化的新的生物學(xué)標(biāo)記物，MIC-1在野生型p53活化時(shí)表達(dá)增高，其他轉(zhuǎn)錄因子如Egr-1、NF-kB均可促進(jìn)MIC-1的促凋亡作用[8]。Koopmann等[5]檢測(cè)胰腺癌、慢性胰腺炎及健康對(duì)照者血清MIC-1水平，發(fā)現(xiàn)MIC-1對(duì)胰腺癌和正常人鑒別診斷較CA19-9更有效。但對(duì)胰腺癌和慢性胰腺炎的鑒別并未表現(xiàn)出優(yōu)勢(shì)。

本研究結(jié)果顯示，胰腺癌患者血清ULBP-2、MIC-1水平顯著高于胰腺癌前病變、慢性胰腺炎患者及健康對(duì)照者。MIC-1水平隨T分期增大而增高。因此MIC-1可能和胰腺癌的進(jìn)展相關(guān)。在胰腺癌診斷方面，單獨(dú)應(yīng)用的效能是ULBP-2>CA19-9>MIC-1，聯(lián)合應(yīng)用可以提高胰腺癌的診斷效能。在診斷胰腺癌早期病變時(shí)，單個(gè)指標(biāo)MIC-1的診斷效能最佳，聯(lián)合應(yīng)用中以MIC-1聯(lián)合CA19-9的診斷效能為最佳。

[1] Goggins M, Canto M, Hruban R. Can we screen high-risk individuals to detect early pancreatic carcinoma? J Surg Oncol, 2000, 74:243-248.

[2] Cosman D, Müllberg J, Sutherland CL, et al. ULBPs, novel MHC class I-related molecules, bind to CMV glycoprotein UL16 and stimulate NK cytotoxicity through the NKG2D receptor. Immunity, 2001, 14: 123-133.

[3] Bootcov MR,Bauskin AR,Valenzuela SM,et al.MIC-1,a novel macrophage inhibitory cytokine,is a divergent member of the TGF-beta superfamily.Proc Natl Acad Sci USA,1997,94:11514-11519.

[4] Chang YT, Wu CC, Shyr YM, et al. Secretome-based identification of ULBP2 as a novel serum marker for pancreatic cancer detection. PLoS One, 2011, 6: e20029.

[5] Koopmann J,Rosenzweig CN,Zhang Z,et al.Serum markers in patients with resectable pancreatic adenocarcinoma:macrophage inhibitory cytokine 1 versus CA19-9.Clin Cancer Res,2006,12:442-446.

[6] Li H, Lakshmikanth T, Garofalo C, et al. Pharmacological activation of p53 triggers anticancer innate immune response through induction of ULBP2. Cell Cycle, 2011, 10: 3346-3358.

[7] Yang H, Filipovic Z, Brown D, et al. Macrophage inhibitory cytokine-1: a novel biomarker for p53 pathway activation. Mol Cancer Ther, 2003, 2: 1023-1029.

[8] Baek SJ, Kim JS, Moore SM, et al. Cyclooxygenase inhibitors induce the expression of the tumor suppressor gene EGR-1, which results in the up-regulation of NAG-1, an antitumorigenic protein. Mol Pharmacol, 2005, 67: 356-364.

DiagnosticvalueofcombinedmeasurementofserumULBP-2andMIC-1forpancreaticcancer

ZHOUYu-fen,HUANGLi-ya,XULing-xiao,ZHANGFan,GUOFang,YAOWei-yan,YUANYao-zong.

DepartmentofGastroenterology,RuijinHospital,ShanghaiJiaotongUniversitySchoolofMedicine,Shanghai200025,China

Correspondingauthor:YUANYao-zong,Email:yyz28@medmail.com.cn

ObjectiveTo investigate the diagnostic value of UL-16 binding protein 2(ULBP-2), macrophage inhibitory cytokine-1(MIC-1) for pancreatic cancer.MethodsThe serum samples of 152 pancreatic cancer patients, 20 precursors of pancreatic cancer, 91 chronic pancreatitis patients and 96 age/sex-matched healthy persons were collected. The serum ULBP-2 and MIC-1 levels were determined by using the ELISA kit and were compared with level of CA19-9. A receiver operating characteristic (ROC) curve was constructed to evaluate their diagnostic values for pancreatic cancer.ResultsThe serum levels of ULBP-2 in patients with pancreatic cancer, precursors of pancreatic cancer, chronic pancreatitis and healthy persons were (219.9±182.5), (62.6±11.4), (68.4±36.8), (76.5±40.9)μg/L, the corresponding values of MIC 1 were (3521.3±3903.4), (973.6±589.0), (959.6±879.0), (427.6±317.0) μg/L, while the corresponding values of CA19-9 were (1448.8±3707.0), (12.0±9.3), (38.2±139.0), (7.7±5.0)kU/L. The parameters in pancreatic cancer patients were significantly higher than those in control group (χ2=40.628,71.662,45.505,15.827,36.433,63.494,26.264,73.427,49.088,P<0.01). The area under ROC curves(AUC) of ULBP-2, MIC-1, CA19-9 were 0.909, 0.864, 0.818, and ULBP-2 was superior to CA19-9 and MIC-1, however the combined measurement of three markers produced the highest diagnostic yield(AUC=0.982). For early stage pancreatic diseases (precursors to pancreatic cancer and IA stage pancreatic cancer), AUC of ULBP-2, MIC-1, CA19-9 were 0.506,0.837,0.684,MIC-1 was superior to ULBP-2 and CA19-9, however the combined measurement of MIC-1 and CA19-9 produced the highest diagnostic yield(AUC=0.897).ConclusionsSerum ULBP-2, MIC-1 levels are significantly elevated in pancreatic cancer patients. The combined measurement of ULBP-2, MIC-1 and CA 19-9 can increase the diagnostic yield for pancreatic cancer.

Pancreatic neoplasms; UL-16 binding proteins 2; Macrophage inhibitory cytokine-1; Biological markers

2012-10-29)

(本文編輯：屠振興)

10.3760/cma.j.issn.1674-1935.2013.02.003

200025 上海，上海交通大學(xué)附屬瑞金醫(yī)院消化內(nèi)科(周郁芬、徐凌霄、張帆、國(guó)芳、姚瑋艷、袁耀宗)；寧夏醫(yī)科大學(xué)總醫(yī)院消化內(nèi)科(黃李雅)

袁耀宗，Email: yyz28@medmail.com.cn