多天线N-糖链对双表型肝细胞癌的辅助诊断价值
DOI: 10.3969/j.issn.1001-5256.2022.06.019
Value of multi-glycan in the auxiliary diagnosis of dual-phenotype hepatocellular carcinoma
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摘要:
目的 探讨双表型肝细胞癌(DPHCC)患者血清中多天线N-糖链(Multi-glycan)的表达及其临床意义。 方法 收集2019年6月—2020年12月福建医科大学孟超肝胆医院65例DPHCC、80例原发性肝细胞癌(HCC)及120例肝硬化(LC)血清样本,采用基于DNA测序仪的荧光毛细管电泳(DSA-FACE)技术检测三组血清中N-糖链的表达,正态分布的计量资料两组间比较采用t检验,多组间比较采用方差分析;非正态分布的计量资料两组间比较采用Mann - Whitney U检验,多组间比较采用Kruskal- Wallis H检验。计数资料组间比较采用χ2检验。采用logistic回归方法建立常见指标模型。采用受试者工作特征(ROC)曲线评价AFP、PIVKA-Ⅱ、CEA、CA19-9及Multi-glycan诊断DPHCC的效能,ROC曲线下面积(AUC)比较采用Z检验。 结果 在DPHCC组及HCC组比较中,仅Multi-glycan差异有统计学意义(P<0.001),而AFP、PIVKA-Ⅱ、CEA、CA19-9及SUM差异均无统计学意义(P值均>0.05)。DPHCC组与HCC组相比,Multi-glycan的曲线下面积(AUCMulti-glycan)为0.775,显著高于AFP(0.507)、PIVKA-Ⅱ(0.584)、CEA(0.537)、CA19-9(0.505)及SUM(0.561),Multi-glycan的灵敏度(69.23%)高于其他5项。在DPHCC组与LC组比较中,Multi-glycan、AFP、PIVKA-Ⅱ、CA19-9及SUM差异均有统计学意义(P值均<0.001)。AUCMulti-glycan(0.780)也均高于AFP(0.767)、PIVKA-Ⅱ(0.743)、CEA(0.566)、CA19-9(0.689)及SUM(0.713),Multi-glycan灵敏度(89.23%)高于其他5项。 结论 Multi-glycan可作为DPHCC的辅助诊断指标之一。 Abstract:Objective To investigate the expression of multi-glycan in serum of patients with dual-phenotype hepatocellular (DPHCC) and its clinical significance. Methods Serum samples were collected from 65 patients with DPHCC, 80 patients with primary hepatocellular carcinoma (HCC), and 120 patients with liver cirrhosis (LC) who were treated in Mengchao Hepatobiliary Hospital of Fujian Medical University from June 2019 to December 2020. DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis was used to measure the expression of N-glycan in serum, The measurement data of normal distribution were compared by t-test between the two groups and analysis of variance between multiple groups; The measurement data with non normal distribution were compared by Mann-Whitney U test between the two groups and Kruskal-Wallis H test between multiple groups, the chi-square test was used for comparison of categorical data between groups.The logistic regression method was used to establish the common index model. The efficacy of AFP, PIVKA - Ⅱ, CEA, CA19-9 and multi glycan in the diagnosis of DPHCC was evaluated by receiver operating characteristic (ROC) curve, and the area under ROC curve (AUC) was compared by Z test. Results There was a significant difference in multi-glycan between the DPHCC group and the HCC group (P < 0.001), while there were no significant differences in AFP, PIVKA-Ⅱ, CEA, CA19-9, and SUM between the two groups (P=0.924, 0.084, 0.442, 0.924, and 0.206). Multi-glycan had an area under the ROC curve (AUC) of 0.775, which was significantly higher than that of AFP (0.507), PIVKA-Ⅱ (0.584), CEA (0.537), CA19-9 (0.505), and SUM (0.561), and multi-glycan had a sensitivity of 69.23%, which was increased compared with the other 5 items. There were significant differences in multi-glycan, AFP, PIVKA-Ⅱ, CA19-9, and SUM between the DPHCC group and the LC group (all P < 0.001), but there was no significant difference in CEA between the two groups (P=0.14). Multi-glycan had an AUC of 0.780, which was also higher than that of AFP (0.767), PIVKA-Ⅱ (0.743), CEA (0.566), CA19-9 (0.689), and SUM (0.713), and multi-glycan had a sensitivity of 89.23%, which was increased compared with the other five items. Conclusion Multi-glycan can be used as one of the indicators for the auxiliary diagnosis of DPHCC. -
Key words:
- Carcinoma, Hepatocellular /
- Multi-antenna of N-glycan /
- Diagnosis
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双表型肝细胞癌(dual-phenotype hepatocellular carcinoma,DPHCC)是2011年首次被报道的肝细胞癌(HCC)的一种新亚型,其发生率约占HCC的10%[1-2]。研究[2]发现,DPHCC患者在性别、年龄、HBV感染、血清甲胎蛋白(AFP)含量、临床分期等方面与HCC患者无明显差异,但DPHCC患者微血管侵犯及肝内外转移的发生率较HCC发生率更高,临床预后更差,复发风险也更高。因此DPHCC的精细分型对于HCC患者的精准治疗及提高临床对HCC亚型的认识具有重要意义。
目前,DPHCC的诊断主要依赖于免疫组化检查,其组织病理学表现为典型的HCC特点,而免疫表型则具有HCC和肝内胆管癌(intrahepatic cholangiocarcinoma,ICC)的双重特性。当肿瘤表现为单一的HCC组织学成分,同时表达任意HCC蛋白标志物(如AFP、HepPar-1、pCEA等)和ICC蛋白标志物(如CK19、CK7、CA19-9等)的癌细胞数量>15%时,即可诊断为DPHCC[3]。免疫组化为诊断DPHCC的金标准。目前常用的DPHCC血清学检测指标有AFP、异常凝血酶原(PIVKA-Ⅱ)、癌胚抗原(CEA)及糖类抗原19-9 (CA19-9)四项,主要为HCC与ICC的辅助诊断指标,尚无特异性DPHCC的血清学指标,因此,需要寻找新的血清学指标用于DPHCC辅助诊断指标。
肝脏参与了血清中近50%糖蛋白中N-糖链的形成。N- 糖链根据天线(或分支)的数量可分为二天线、三天线、四天线及四天线以上N-糖链结构,其中三天线、四天线及四天线以上N-糖链结构称为多天线N-糖链结构(Multi-glycan)[4]。研究[5]发现血清N-糖链表达水平(量)与糖链结构(质)的改变可以成为HCC潜在诊断标志物或治疗靶标,特别是血清中Multi-glycan表达水平的变化与肿瘤的复发、转移等密切相关。Huang等[6]研究发现,HCC患者血清三天线N-糖链结构丰度显著增加,同时复发患者三天线及四天线丰度显著增加,部分复发患者甚至出现四天线以上N-糖链结构,Multi-glycan可以作为HCC患者复发的潜在标志物之一。对于复发风险更高的DPHCC而言,患者血清中Multi-glycan的表达情况目前尚无相关文献报道。因此,本研究拟通过比较DPHCC中常见血清肿瘤指标与Multi-glycan的表达,探讨Multi-glycan对DPHCC的辅助诊断价值。
1. 资料与方法
1.1 研究对象
收集2019年6月—2020年12月福建医科大学孟超肝胆医院220例患者血清,分为DPHCC组、HCC组和肝硬化组。收集所有患者的一般临床资料、实验室检测资料及病理资料。HCC诊断依据《原发性肝癌诊疗规范(2017年版)》[7]。DPHCC诊断依据《原发性肝癌诊疗规范(2019年版)》[8]。LC诊断依据《慢性乙型肝炎防治指南(2019年版)》[9]。
1.2 纳入及排除标准
1.2.1 HCC及DPHCC组
HCC及DPHCC均为经术后病理检查明确诊断。排除标准:(1)继发性肝癌、其他器官恶性肿瘤、严重感染及其他器官严重疾病等;(2) HBV之外的其他病毒感染,如HAV、HCV等;(3)其他慢性肝病引起的肝癌,如:酒精性脂肪肝、自身免疫性肝病等。
1.2.2 LC组
LC均经过临床表现和影像学检查确诊。排除标准:(1)HBV之外的其他病毒感染,如HAV、HCV等;(2)其他原因引起的肝硬化,如原发性胆汁性肝硬化、酒精性肝硬化、血吸虫肝硬化等。
1.3 检测方法
采用基于DNA测序仪的荧光毛细管电泳(DNA sequence-assisted fluorophore-assisted capillary electrophoresis, DSA-FACE)技术,通过N-糖链释放、荧光标记、去除唾液酸及测序仪检测四个步骤,进行N-糖链结构分析检测,具体操作步骤见参考文献[10]。
1.4 实验室常规指标检测
收集所有对象的实验室检测资料,包括AFP、PIVKA-Ⅱ、CEA、CA19-9、TP、Alb、TBil、ALT及AST。其中AFP、PIVKA-Ⅱ、CEA及CA19-9四项均采用LLMIPULSEG1200全自动化学发光仪(日本富士公司)及配套试剂检测。TP、Alb、TBil、ALT及AST五项均采用AU5800全自动生化分析仪(贝克曼公司)及配套试剂检测。
1.5 统计学方法
采用SPSS23.0软件进行统计学分析。正态分布计量资料以x±s表示,两组间比较采用t检验,多组间比较采用方差分析;非正态分布计量资料以M(P25~P75)表示,两组间比较采用Mann - Whitney U检验,多组间比较采用Kruskal - Wallis H检验。计数资料组间比较采用χ2检验。采用logistic回归方法建立常见指标模型。采用受试者工作特征曲线(ROC曲线)评价AFP、PIVKA-Ⅱ、CEA、CA19-9及Multi-glycan诊断DPHCC的效能,ROC曲线下面积(AUC)比较采用Z检验。P<0.05为差异有统计学意义。
2. 结果
2.1 一般资料
DPHCC组65例,其中男55例、女10例,年龄(55.00±16.50)岁;HCC组80例,其中男67例、女13例,年龄(56.50±16.00)岁;肝硬化组120例,其中男98例、女22例,年龄(55.00±19.00)岁。
2.2 血清总N-糖链结果
DSA-FACE检测可将血清总N-糖链分为13个峰(paek1~peak12及peak9’)。各峰表示的N-糖链结构如图 1所示,Peak1:NGA2F, 无半乳糖-α-1, 6-核心岩藻糖基化二天线结构;Peak2:NGA2FB, 无半乳糖-α-1, 6-核心岩藻糖基化平分型二天线结构;Peak3/Peak4:NG1A2F, 单半乳糖-α-1, 6-核心岩藻糖基化二天线结构;Peak5:NA2, 双半乳糖二天线结构;Peak6:NA2F, 双半乳糖-α-1, 6-核心岩藻糖基化二天线结构;Peak7:NA2FB, 双半乳糖-α-1, 6-核心岩藻糖基化平分型二天线结构;Peak8:NA3, 三半乳糖三天线结构;Peak9:NA3Fb, 三半乳糖-α-1, 3-分支岩藻糖基化三天线结构;Peak10:NA3Fc, 三半乳糖-α-1, 6-核心岩藻糖基化-α-1, 3-分支岩藻糖基化三天线结构;Peak11:NA4, 四半乳糖四天线结构;Peak12:NA4Fb, 四半乳糖α-1, 3-分支岩藻糖基化四天线结构。其中Peak1~Peak7为二天线N-糖链结构,Peak8、Peak9、Peak9’及Peak10为三天线N-糖链结构,peak11及peak12为四天线N-糖链结构,Peak8、Peak9、Peak9’、Peak10、Peak11及Peak12即为多天线N-糖链结构。
2.3 三组中各指标的表达水平
DPHCC组、HCC组及LC组之间性别及年龄差异均无统计学意义(P值均>0.05);肝功能指标中TP、Alb、ALT及AST差异均有统计学意义(P值均<0.05),TBil差异无统计学意义(P>0.05);血清肿瘤指标中AFP、PIVKA-Ⅱ、CEA及CA19-9差异均有统计学意义(P值均<0.05);采用logictic回归方法构建常见指标模型(SUM,SUM=0.63×AFP+0.021×PIVKA-Ⅱ+0.01×CA19-9-0.006×CEA+0.037),SUM在3组中差异也具有统计学意义(P<0.05)(表 1)。13个N-糖链中,peak1、peak2、peak7、peak8及peak12差异均无统计学意义(P值均>0.05),而peak3、peak4、peak5、peak6、peak9、peak9’、peak10、peak11及Multi-glycan差异均有统计学意义(P值均<0.05),如表 1所示。
表 1 三组中各指标的表达水平Table 1. The expression of various indexes in three groups指标 DPHCC组(n=65) HCC组(n=80) LC组(n=120) 统计值 P值 男性[例(%)] 55 (84.62) 67 (83.75) 98 (83.05) χ2=0.03 0.975 年龄(岁) 55.00(46.50~63.00) 56.50(49.00~64.75) 55.00(46.25~65.00) H=0.17 0.727 AFP (ng/mL) 59.84(9.25~906.32) 73.28(5.95~2 000.00) 6.65(3.50~26.90) H=10.61 <0.001 PIVKA-Ⅱ (mAU/mL) 154.00(40.00~865.50) 603.00(50.50~5 940.14) 38.50(32.25~48.25) H=9.90 <0.001 CEA (ng/mL) 2.70(1.60~4.30) 3.00(1.80~4.60) 2.55(1.50~2.55) H=4.19 0.044 CA19-9 (U/mL) 16.84(8.96~31.72) 18.88(8.17~31.38) 44.86(13.57~73.84) H=5.19 <0.001 TP (g/L) 64.51±9.10 63.56±9.63 68.94±7.63 F=11.02 <0.001 Alb (g/L) 37.60±5.17 36.78±6.00 40.01±5.23 F=9.50 <0.001 TBil (μmol/L) 18.20(13.15~23.85) 17.10(12.88~22.78) 16.80(12.00~28.55) H=2.80 0.06 ALT (U/L) 53.00(33.50~101.00) 69.00(33.25~188.50) 32.00(23.25~64.75) H=14.38 <0.001 AST (U/L) 57.00 (32.00~113.00) 79.00(37.25~264.25) 31.00(23.00~49.75) H=16.30 <0.001 SUM 398.80(224.40~712.00) 377.50(169.10~622.40) 897.20(271.40~1 457.80) H=4.74 0.01 肿瘤分期(例) Ⅰ级 0 0 Ⅱ级 20 25 Ⅲ级 40 48 Ⅳ级 5 7 N-glycan Peak1 8.78(6.51~10.52) 8.64(6.93~10.20) 8.62(6.52~8.62) H=0.33 0.652 Peak2 1.27(0.95~1.89) 1.31(1.04~1.69) 1.15(0.79~1.77) H=1.02 0.216 Peak3 6.69(5.65~8.01) 6.08(5.41~7.12) 7.24(6.27~8.72) H=12.20 <0.001 Peak4 5.92±0.90 5.72±0.86 5.27±0.97 F=12.10 <0.001 Peak5 38.26±5.52 39.40±5.24 34.29±8.39 F=15.07 <0.001 Peak6 20.37±3.10 18.74±3.39 22.61±5.37 F=19.63 <0.001 Peak7 6.19±1.38 5.99±1.45 6.58±2.17 F=2.75 0.172 Peak8 6.81±2.22 6.13±2.01 6.17±2.62 F=0.98 0.241 Peak9 4.18(2.92~4.18) 3.24(2.17~5.15) 2.82(2.20~4.31) H=5.97 0.001 Peak9’ 0.98(0.80~1.16) 0.76(0.63~1.00) 0.92(0.70~1.29) H=5.60 0.001 Peak10 0.16(0.09~0.69) 0.30(0.23~0.37) 0.38(0.26~0.57) H=5.62 0.001 Peak11 1.99±0.62 1.64±0.64 1.59±0.73 F=7.87 0.001 Peak12 0.86(0.64~1.02) 0.65(0.49~1.11) 0.70(0.58~0.94) H=3.36 0.073 Multi-glycan 16.87±2.96 13.60±3.30 13.32±3.58 F=25.93 <0.001 注:Multi-glycan= Peak8+Peak9+Peak9’+Peak10+Peak11+Peak12。 2.4 三组中血清肿瘤指标及Multi-glycan的表达
DPHCC组与HCC组比较,仅Multi-glycan差异有统计学意义(P<0.05),而AFP、PIVKA-Ⅱ、CEA、CA19-9及SUM差异均无统计学意义(P值均>0.05);DPHCC组与LC组比较,Multi-glycan、AFP、PIVKA-Ⅱ、CA19-9及SUM差异均有统计学意义(P值均<0.001),而CEA差异无统计学意义(P=0.14)(图 2)。
2.5 三组中血清肿瘤指标、差异N-糖链及Multi-glycan的AUC比较
DPHCC组与HCC组比较,各指标诊断DPHCC的AUC分别为:AUCMulti-glycan为0.775(95%CI: 0.699~0.850)、AUCpeak3为0.596(95%CI: 0.502~0.691)、AUCpeak4为0.566(95%CI: 0.472~0.660)、AUCpeak5为0.559(95%CI: 0.465~0.653)、AUCpeak6为0.656(95%CI: 0.567~0.746)、AUCpeak9为0.583(95%CI: 0.489~0.677)、AUCpeak9’为0.685(95%CI: 0.599~0.772)、AUCpeak10为0.570(95%CI: 0.457~0.683)、AUCpeak11为0.655(95%CI: 0.566~0.744)、AUCAFP为0.507(95%CI: 0.699~0.850)、AUCPIVKA-Ⅱ为0.584(95%CI: 0.490~0.677)、AUCCEA为0.537(95%CI: 0.442~0.632)、AUCCA19-9为0.505(95%CI: 0.410~0.600)及AUCSUM为0.561(95%CI: 0.467~0.655)。其中,AUCMulti-glycan显著高于AUCpeak3(Z=4.96, P<0.001)、AUCpeak4(Z=4.81, P<0.001)、AUCpeak5(Z=4.73, P<0.001)、AUCpeak6(Z=2.97, P=0.003)、AUCpeak9(Z=3.79, P<0.001)、AUCpeak10(Z=3.19, P<0.001)、AUCpeak11(Z=3.10, P<0.001)、AUCAFP(Z=4.62, P<0.001)、AUCPIVKA-Ⅱ(Z=3.30, P=0.001)、AUCCEA(Z=3.99, P<0.001)、AUCCA19-9(Z=4.49, P<0.001)及AUCSUM(Z=3.60, P<0.001),差异均有统计学意义(图 3,表 2)。
表 2 三组中各指标的诊断效能Table 2. The diagnostic efficiency of various indexes in three groups指标 Cut-off值 灵敏度(%) 特异度(%) AUC(95%CI) DPHCC组vs HCC组 AFP 6.67 ng/mL 35.38 73.75 0.507(0.413~0.601) PIVKA-Ⅱ 554 mAU/mL 27.69 41.25 0.584(0.490~0.677) CEA 1.65 ng/mL 58.46 22.50 0.537(0.442~0.632) CA19-9 23.59 U/mL 33.85 53.75 0.505(0.410~0.600) SUM 690.40 69.23 56.25 0.561(0.467~0.655) Multi-glycan 16.67 69.23 73.75 0.775(0.699~0.850) DPHCC组vs LC组 AFP 106.13 ng/mL 30.77 74.17 0.767(0.690~0.844) PIVKA-Ⅱ 72.50 mAU/mL 64.62 75.80 0.743(0.645~0.841) CEA 3.85 ng/mL 32.31 65.00 0.566(0.476~0.656) CA19-9 43.48 U/mL 12.31 46.67 0.689(0.610~0.767) SUM 865.80 71.88 45.83 0.713(0.642~0.784) Multi-glycan 13.51 89.23 66.67 0.780(0.713~0.848) DPHCC组与LC组比较,各指标诊断DPHCC的AUC分别为:AUCMulti-glycan为0.780(95%CI: 0.713~0.848)、AUCpeak3为0.699(95%CI: 0.625~0.772)、AUCpeak4为0.629(95%CI: 0.552~0.706)、AUCpeak5为0.690(95% CI: 0.616~0.763)、AUCpeak6为0.760(95%CI: 0.704~0.806)、AUCpeak9为0.564(95%CI: 0.479~0.649)、AUCpeak9’为0.606(95%CI: 0.527~0.685)、AUCpeak10为0.631(95%CI: 0.555~0.707)、AUCpeak11为0.508(95% CI: 0.428~0.589)、AUCAFP为0.767(95%CI: 0.691~0.844)、AUCPIVKA-Ⅱ为0.743(95%CI: 0.645~0.841)、AUCCEA为0.537(95%CI: 0.476~0.656)、AUCCA19-9为0.689(95%CI: 0.625~0.779)及AUCSUM为0.713(95%CI: 0.628~0.788)。其中,AUCMulti-glycan显著高于AUCpeak3(Z=5.65, P<0.001)、AUCpeak4(Z=2.40, P=0.01)、AUCpeak5(Z=3.93, P<0.001)、AUCpeak6(Z=3.40, P<0.001)、AUCpeak9(Z=2.28, P=0.02)、AUCpeak9’(Z=5.31, P<0.001)、AUCpeak10(Z=2.59, P=0.009)、AUCpeak11(Z=4.14, P<0.001)及AUCCEA (Z=3.87, P<0.001),差异均有统计学意义(图 3,表 2)。
2.6 三组中血清肿瘤指标及Multi-glycan的诊断效能分析
DPHCC组与HCC组比较,Multi-glycan的灵敏度与SUM相同均为69.23%,且比AFP、PIVKA-Ⅱ、CEA、CA19-9分别提高33.85%、41.54%、10.77%、35.38%;Multi-glycan的特异度与AFP相同均为73.75%,且比PIVKA-Ⅱ、CEA、CA19-9、SUM分别提高32.5%、51.25%、20%、17.5%。
DPHCC组与LC组比较中,Multi-glycan的灵敏度为89.23%,且比AFP、PIVKA-Ⅱ、CEA、CA19-9、SUM分别提高58.46%、24.61%、56.92%、76.92%、17.35%;Multi-glycan特异度为66.67%,且比CEA、CA19-9、SUM提高1.67%、20%、20.84%,而稍低于AFP、PIVKA-Ⅱ(表 2)。
3. 讨论
DPHCC是近几年新发现的HCC新亚型,其总生存期和无复发生存期显著低于普通型HCC患者[1]。因此,DPHCC的早期诊断对于改善患者的诊疗现状、提高患者的预后生存具有重要意义。
N-糖链作为蛋白质翻译后糖基化修饰的产物之一,近年来已成为无创性肿瘤标志物的研究方向之一,可以作为胃癌、乳腺癌、膀胱癌、肺癌等肿瘤的血清学辅助诊断指标[11-13]。N-糖链主要通过蛋白Asn-XXX-Ser/Thr序列(XXX为除脯氨酸以外的氨基酸)中Asn残基上的-NH2与蛋白相连[14-15]。多数糖蛋白的N-糖链为二或三天线(或分支)结构,少数为四天线结构,四天线以上较为罕见。其中三天线及以上天线结构的N-糖链称为Multi-glycan。
本研究结果显示,HCC组与DPHCC组中四天线N-糖链结构(peak11及peak12)及Multi-glycan的丰度均高于LC组,与Huang等[6]文献报道相似。同时,DPHCC组中Multi-glycan的丰度也显著高于HCC组(P<0.05),提示Multi-glycan的丰度可能可用于DPHCC患者的血清学诊断。
DPHCC组与LC组ROC曲线分析结果显示,DPHCC组中Multi-glycan的AUC不仅高于各单项差异N-糖链结构的AUC(peak3、peak4、peak5、peak6、peak9、peak9’、peak10、peak11),也显著高于AFP、PIVKA-Ⅱ、CEA、CA19-9及SUM模型。Multi-glycan的灵敏度(89.23%)也均高于四项血清肿瘤指标及SUM模型。DPHCC组与HCC组ROC曲线分析结果显示,DPHCC组中Multi-glycan的AUC也显著高于各单项差异N-糖链结构、四项血清肿瘤指标及SUM模型,且Multi-glycan的灵敏度(69.23%)与SUM模型相同,均高于四项血清肿瘤指标。诊断效能分析结果进一步表明,Multi-glycan不仅能区分DPHCC与LC患者,也能很好的区分DPHCC与HCC患者。综上所述,Multi-glycan可以作为DPHCC的血清学辅助诊断指标。
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表 1 三组中各指标的表达水平
Table 1. The expression of various indexes in three groups
指标 DPHCC组(n=65) HCC组(n=80) LC组(n=120) 统计值 P值 男性[例(%)] 55 (84.62) 67 (83.75) 98 (83.05) χ2=0.03 0.975 年龄(岁) 55.00(46.50~63.00) 56.50(49.00~64.75) 55.00(46.25~65.00) H=0.17 0.727 AFP (ng/mL) 59.84(9.25~906.32) 73.28(5.95~2 000.00) 6.65(3.50~26.90) H=10.61 <0.001 PIVKA-Ⅱ (mAU/mL) 154.00(40.00~865.50) 603.00(50.50~5 940.14) 38.50(32.25~48.25) H=9.90 <0.001 CEA (ng/mL) 2.70(1.60~4.30) 3.00(1.80~4.60) 2.55(1.50~2.55) H=4.19 0.044 CA19-9 (U/mL) 16.84(8.96~31.72) 18.88(8.17~31.38) 44.86(13.57~73.84) H=5.19 <0.001 TP (g/L) 64.51±9.10 63.56±9.63 68.94±7.63 F=11.02 <0.001 Alb (g/L) 37.60±5.17 36.78±6.00 40.01±5.23 F=9.50 <0.001 TBil (μmol/L) 18.20(13.15~23.85) 17.10(12.88~22.78) 16.80(12.00~28.55) H=2.80 0.06 ALT (U/L) 53.00(33.50~101.00) 69.00(33.25~188.50) 32.00(23.25~64.75) H=14.38 <0.001 AST (U/L) 57.00 (32.00~113.00) 79.00(37.25~264.25) 31.00(23.00~49.75) H=16.30 <0.001 SUM 398.80(224.40~712.00) 377.50(169.10~622.40) 897.20(271.40~1 457.80) H=4.74 0.01 肿瘤分期(例) Ⅰ级 0 0 Ⅱ级 20 25 Ⅲ级 40 48 Ⅳ级 5 7 N-glycan Peak1 8.78(6.51~10.52) 8.64(6.93~10.20) 8.62(6.52~8.62) H=0.33 0.652 Peak2 1.27(0.95~1.89) 1.31(1.04~1.69) 1.15(0.79~1.77) H=1.02 0.216 Peak3 6.69(5.65~8.01) 6.08(5.41~7.12) 7.24(6.27~8.72) H=12.20 <0.001 Peak4 5.92±0.90 5.72±0.86 5.27±0.97 F=12.10 <0.001 Peak5 38.26±5.52 39.40±5.24 34.29±8.39 F=15.07 <0.001 Peak6 20.37±3.10 18.74±3.39 22.61±5.37 F=19.63 <0.001 Peak7 6.19±1.38 5.99±1.45 6.58±2.17 F=2.75 0.172 Peak8 6.81±2.22 6.13±2.01 6.17±2.62 F=0.98 0.241 Peak9 4.18(2.92~4.18) 3.24(2.17~5.15) 2.82(2.20~4.31) H=5.97 0.001 Peak9’ 0.98(0.80~1.16) 0.76(0.63~1.00) 0.92(0.70~1.29) H=5.60 0.001 Peak10 0.16(0.09~0.69) 0.30(0.23~0.37) 0.38(0.26~0.57) H=5.62 0.001 Peak11 1.99±0.62 1.64±0.64 1.59±0.73 F=7.87 0.001 Peak12 0.86(0.64~1.02) 0.65(0.49~1.11) 0.70(0.58~0.94) H=3.36 0.073 Multi-glycan 16.87±2.96 13.60±3.30 13.32±3.58 F=25.93 <0.001 注:Multi-glycan= Peak8+Peak9+Peak9’+Peak10+Peak11+Peak12。 表 2 三组中各指标的诊断效能
Table 2. The diagnostic efficiency of various indexes in three groups
指标 Cut-off值 灵敏度(%) 特异度(%) AUC(95%CI) DPHCC组vs HCC组 AFP 6.67 ng/mL 35.38 73.75 0.507(0.413~0.601) PIVKA-Ⅱ 554 mAU/mL 27.69 41.25 0.584(0.490~0.677) CEA 1.65 ng/mL 58.46 22.50 0.537(0.442~0.632) CA19-9 23.59 U/mL 33.85 53.75 0.505(0.410~0.600) SUM 690.40 69.23 56.25 0.561(0.467~0.655) Multi-glycan 16.67 69.23 73.75 0.775(0.699~0.850) DPHCC组vs LC组 AFP 106.13 ng/mL 30.77 74.17 0.767(0.690~0.844) PIVKA-Ⅱ 72.50 mAU/mL 64.62 75.80 0.743(0.645~0.841) CEA 3.85 ng/mL 32.31 65.00 0.566(0.476~0.656) CA19-9 43.48 U/mL 12.31 46.67 0.689(0.610~0.767) SUM 865.80 71.88 45.83 0.713(0.642~0.784) Multi-glycan 13.51 89.23 66.67 0.780(0.713~0.848) -
[1] ZHENG QL, FENG CY, LIAN YE, et al. Clinical and pathological analysis of 6 cases of diphenotypic hepatocellular carcinoma[J]. Chin J Pathol, 2020, 49(12): 1320-1322. DOI: 10.3760/cma.j.cn112151-20200319-00237.郑巧灵, 冯昌银, 连渊娥, 等. 双表型肝细胞癌六例临床病理学分析[J]. 中华病理学杂志, 2020, 49(12): 1320-1322. DOI: 10.3760/cma.j.cn112151-20200319-00237. [2] WANG H, CONG WM. Research progress on clinicopathology in dual-phenotype hepatocellular carcinoma[J]. Chin J Clin Oncol, 2017, 44(12): 616-619. DOI: 10.3969/j.issn.1000-8179.2017.12.071.王瀚, 丛文铭. 双表型肝细胞癌新亚型的临床病理学研究进展[J]. 中国肿瘤临床, 2017, 44(12): 616-619. DOI: 10.3969/j.issn.1000-8179.2017.12.071. [3] Chinese Society of Liver Cancer, Chinese Anti-Cancer Association, Liver Cancer Study Group, Chinese Society of Hepatology, Chinese Medical Association, Chinese Society of Pathology, Chinese Anti-Cancer Association, et al. Evidence-based practice guidelines for the standardized pathological diagnosis of primary liver cancer in China(2015 update)[J]. J Clin Hepatol, 2015, 31(6): 833-839. DOI: 10.3969/j.issn.1001-5256.2015.06.004.中国抗癌协会肝癌专业委员会, 中华医学会肝病学分会肝癌学组, 中国抗癌协会病理专业委员会, 等. 原发性肝癌规范化病理诊断指南(2015年版)[J]. 临床肝胆病杂志, 2015, 31(6): 833-839. DOI: 10.3969/j.issn.1001-5256.2015.06.004. [4] FANG M, GAO CF. Glycosylation and biomarkers in primary liver cancer[J]. Chin J Lab Med, 2016, 39(2): 73-75. DOI: 10.3760/cma.j.issn.1009-9158.2016.02.001.房萌, 高春芳. 糖基化与原发性肝癌标志物[J]. 中华检验医学杂志, 2016, 39(2): 73-75. DOI: 10.3760/cma.j.issn.1009-9158.2016.02.001. [5] HUANG C, FANG M, FENG H, et al. N-glycan fingerprint predicts alpha-fetoprotein negative hepatocellular carcinoma: A large-scale multicenter study[J]. Int J Cancer, 2021, 149(3): 717-727. DOI: 10.1002/ijc.33564. [6] HUANG C, LIU L, WANG H, et al. Serum N-glycan fingerprint nomogram predicts liver fibrosis: A multicenter study[J]. Clin Chem Lab Med, 2021, 59(6): 1087-1097. DOI: 10.1515/cclm-2020-1588. [7] National Health and Family Planning Commission of the People's Republic of China. Diagnosis, management, and treatment of hepatocellular carcinoma (V2017)[J]. J Clin Hepatol, 2017, 33(8): 1419-1431. DOI: 10.3969/j.issn.1001-5256.2017.08.003.中华人民共和国国家卫生和计划生育委员会. 原发性肝癌诊疗规范(2017年版)[J]. 临床肝胆病杂志, 2017, 33(8): 1419-1431. DOI: 10.3969/j.issn.1001-5256.2017.08.003. [8] Bureau of Medical Administration, National Health Commission of the People's Republic of China. Guidelines for diagnosis and treatment of primary liver cancer in China (2019 edition)[J]. J Clin Hepatol, 2020, 36(2): 277-292. DOI: 10.3969/j.issn.1001-5256.2020.02.007.中华人民共和国国家卫生健康委员会医政医管局. 原发性肝癌诊疗规范(2019年版)[J]. 临床肝胆病杂志, 2020, 36(2): 277-292. DOI: 10.3969/j.issn.1001-5256.2020.02.007. [9] Chinese Society of Infectious Diseases, Chinese Medical Association, Chinese Society of Hepatology, Chinese Medical Association. Guidelines for the prevention and treatment of chronic hepatitis B (version 2019)[J]. J Clin Hepatol, 2019, 35(12): 2648-2669. DOI: 10.3969/j.issn.1001-5256.2019.12.007.中华医学会感染病学分会, 中华医学会肝病学分会. 慢性乙型肝炎防治指南(2019年版)[J]. 临床肝胆病杂志, 2019, 35(12): 2648-2669. DOI: 10.3969/j.issn.1001-5256.2019.12.007. [10] GUAN W, GAO Z, HUANG C, et al. The diagnostic value of serum DSA-TRF in hepatocellular carcinoma[J]. Glycoconj J, 2020, 37(2): 231-240. DOI: 10.1007/s10719-019-09906-x. [11] LI T, MO C, QIN X, et al. Glycoprofiling of early gastric cancer using lectin microarray technology[J]. Clin Lab, 2018, 64(1): 153-161. DOI: 10.7754/Clin.Lab.2017.170814. [12] WANG Z, LIU H, YAN Y, et al. Integrated proteomic and N-Glycoproteomic analyses of human breast cancer[J]. J Proteome Res, 2020, 19(8): 3499-3509. DOI: 10.1021/acs.jproteome.0c00311. [13] TANAKA T, YONEYAMA T, NORO D, et al. Aberrant N-Glycosylation profile of serum immunoglobulins is a diagnostic biomarker of urothelial carcinomas[J]. Int J Mol Sci, 2017, 18(12): 2632. DOI: 10.3390/ijms18122632. [14] BEDNARSKA NG, WREN BW, WILLCOCKS SJ. The importance of the glycosylation of antimicrobial peptides: natural and synthetic approaches[J]. Drug Discov Today, 2017, 22(6): 919-926. DOI: 10.1016/j.drudis.2017.02.001. [15] MARTINEZ MR, DIAS TB, NATOV PS, et al. Stress-induced O-GlcNAcylation: An adaptive process of injured cells[J]. Biochem Soc Trans, 2017, 45(1): 237-249. DOI: 10.1042/BST20160153. -