肝细胞癌肿瘤标志物的新进展

肝细胞癌(HCC)是最常见的恶性肿瘤之一,在中国其病死率居恶性肿瘤的第三位.肿瘤标志物的测定是早期诊断和监测复发的有效方法 ,近年来这方面的研究进展迅速,它可包括癌胚抗原和糖类抗原、酶和同工酶、基因和细胞因子等四大类.本文综述了它们在HCC诊断、疗效评价、预后判断以及复发监测等方面的研究进展,并介绍了一些近年新发现的一些肿瘤标志物,如白介素6、LAM4基因、NDRG1基因等.     

肝细胞癌的相关血清标志物

肝细胞癌(HCC)的早期诊断是其治疗的关键,HCC血清标志物的检测为其诊断提供了有利的方法,并且操作简单,敏感性高、特异性强。目前常用的血清标志物为AFP、AFPvariants、AFPmRNA、AFU、GGT、DCP、AIF、GPC3等。这些标志物的联合使用有助于HCC的诊断及预后判断。     

肝细胞癌的相关血清标志物

HCC的早期诊断是其治疗的关键，HCC血清标志物的检测又为其诊断提供了有利的途径，并且操作简单，敏感性高和特异性强。目前常用的血清标志物为AFP、AFP变异体、AFP mRNA、AFU、GGT、DCP、AIF、GPC3等。这些标志物的联合使用有助于HCC的诊断及预后。     

肝癌与肝炎标志物及相关肿瘤标志物的相关性分析

目的探讨乙肝五项、HCV抗体、血清甲胎蛋白(AFP)、CA19-9、CA125、CEA对原发性肝癌的诊断价值。方法对75例原发性肝癌患者分别测定乙肝五项、HCV抗体、肝脏肿瘤血清标志物AFP、CA199、CA125、CEA。结果 5例原发性肝癌患者中,HBsAg阳性45例(60.0%),HCV抗体阳性16例(21.3%),HBsAg并HCV抗体双阳性3例(4.0%);乙肝五项(HBV-M)不同组合模式中,HBsAg、HBeAb、HBcAb三项阳性(小三阳)20例,HBsAg、HBeAg、HBcAb阳性(大三阳)9例,HBsAg、HBcAb两项阳性14例。AFP、CA19-9、CA125、CEA对肝癌诊断的灵敏度依次为AFP(68.0%),CA19-9(34.6%),CA125(33.3%),CEA(30.7%),四项联合检测对肝癌诊断的灵敏度可提高至80.0%。结论肝癌的发生与HBV有着非常密切的关系,HBsAg、HBeAb、HBcAb三项阳性(小三阳)患者应视为的高危人群。在肝癌患者中,肝癌的单项肿瘤标志物检测应首选阳性率最高的AFP,多种肿瘤标志物联合检测对肝癌的诊断具有重要价值。     

肝细胞生长因子及其受体与肝细胞癌

肝细胞生长因子(HGF)是正常肝细胞增生的强促有丝分裂原。近年研究发现，HGF同样影响肝细胞癌的生物学行为，其受体(c-Met原癌基因编码蛋白)也与原发性肝癌关系密切，因此可能成为原发性肝癌新的诊断标志物和治疗靶点。     

异常凝血酶原和肝细胞癌

目前肝细胞癌（hepatocellular carcinoma,HCC)的诊断主要有影像学诊断和血清肿瘤标志物的检测。异常凝血酶原（des-gamma-carboxy-prothrombin,DCP)又被称为PIVKA－Ⅱ（protein induced by vitamin K absence or antagonist-Ⅱ），与AFP（alpha-fetoprotein)和AFP－L3（alpha-fetoprotein L3 fraction)一样被认为是一种很有价值的肝细胞癌血清肿瘤标志物。在HCC的检测诊断上，它们之间无明显相关关系，而表现为一定的互补性，结合影像学诊断，动态观测HCC高危（肝炎、肝硬化）人群，这些血清肿瘤标志物有助于HCC的早期发现，同时对HCC的手术疗效的评价、预后的估评有着一定的指导意义。     

肝癌的新型肿瘤标志物

肝细胞癌是全球最常见的恶性肿瘤之一,并在最常见的肿瘤致死原因中位列第三。随着医学的发展,肝癌的诊疗手段已经取得明显的进步,但由于缺乏有效的早期诊断方式,往往使得早期肝癌患者无法得到及时的治疗。因此,开发有效的肝癌早期诊断手段也将是一个改善患者预后的合理途径。本文通过系统复习相关文献,总结了肝癌部分常用肿瘤标志物(如甲胎蛋白)以及新型肿瘤标志物的机制及意义。     

生长激素受体mRNA在肝细胞癌和癌旁肝硬化组织中的表达

背景与目的:正常肝组织富含生长激素受体mRNA,但在肝细胞癌、癌旁肝硬化组织中生长激素受体mRNA的表达情况不详,因此,本文旨在探讨肝癌组织、癌旁肝硬化组织中生长激素受体mRNA的表达情况。方法:采用逆转录-聚合酶链反应方法检测37例肝癌患者肝癌组织、癌旁肝硬化组织中生长激素受体mRNA的表达。结果:肝癌组织生长激素受体mRNA表达率(30/37,81.0%)较癌旁肝硬化组织的表达率(32/32,100%)低(P<0.05),未分化肝癌的表达率(1/4,20.0%)更低(P<0.5);癌旁肝硬化组织的生长激素受体mRNA的表达量犤积分光密度比值(riOD)为(30.77±8.24)%,n=32犦较正常对照肝组织的表达量犤riOD为(44.93±6.25)%,n=5犦少(P<0.05),重度肝硬化的表达量犤riOD为(21.90±4.72)%,n=8犦更少(P<0.05)。结论:生长激素受体mRNA的表达下调与肝癌的分化程度、癌旁肝硬化的严重程度有关。     

Risk Scoring System based on lncRNA Expression for Predicting Survival in Hepatocellular Carcinoma with Cirrhosis

Objective: This study aims to explore the roles of long non-coding RNAs (lncRNAs) for predicting survival in hepatocellular carcinoma (HCC) patients with cirrhosis. Methods: A set of lncRNAs differentially expressed between HCC patients with or without cirrhosis was identified using expression profiles of The Cancer Genome Atlas database, and these lncRNAs were screened for their risk scoring system to predict recurrence-free survival (RFS) or overall survival (OS). Predictive ability of risk scoring systems was confirmed using uni- and multivariate Cox analyses while adjusting for clinical features. Predictive lncRNAs were analyzed by functional enrichment analysis. Results: Our screen identified 22 lncRNAs that were upregulated in the presence of cirrhosis and 59 that were downregulated. To predict OS of HCC patients with cirrhosis, a risk scoring system was developed with four lncRNAs (LINC02086, LINC00880, LINC01549 and AC136475.3); to predict RFS in these patients, the risk scoring system contained five lncRNAs (SH3RF3-AS1, AC104117.3, AC136475.3, LINC00239 and MRPL23-AS1). All risk scoring systems were associated with an area under the receiver operating characteristic curve > 0.7. Based on uni- and multivariate Cox analyses, the risk scoring system could serve as a significant independent predictor for OS in HCC patients with cirrhosis. Functional enrichment analysis suggested that the lncRNAs in the risk scoring systems are involved primarily in the pathway of Wnt signal and cytokine-cytokine receptor interaction. Conclusion: Risk scoring systems based on lncRNAs can effectively predict OS of HCC patients with cirrhosis. The system should be further developed and validated in larger, preferably multi-site patient populations.     

Bicluster and Pathway Enrichment Analysis of HCV-induced Cirrhosis and Hepatocellular Carcinoma

Background: Hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide and the mostcommon form of liver cancer. However, while it is associated frequently with hepatitis C virus (HCV) there isonly an elementary understanding of its molecular pathogenesis. Methods: To gain insight into the molecularmechanisms of HCV-induced hepatocarcinogenesis, we performed microarray analysis on 75 surgical liversamples from 48 HCV-infected patients. Results: There were 395 differentially expressed geness between cirrhoticsamples and HCC samples. Of these, 125 genes were up-regulated and 270 genes were down-regulated. Weperformed pathway enrichment analysis and screened as described previously. Conclusions: The differentiallyexpressed genes might be involved in hepatocarcinogenesis through upregulating the pathways of ECM-receptorinteraction, focal adhesion, cell adhesion molecules and other cancer-related pathways, and downregulating thepathways of “complement and coagulation cascades”. We hope our results could aid in seeking of therapeutictargets for HCV-induced hepatocellular carcinoma.     

Expression of Matrilin-2 in Liver Cirrhosis and Hepatocellular Carcinoma

The recently described matrilin protein family is part of the extracellular matrix, their pathophysiological role as well as distribution in liver diseases, however, have not yet been studied. Considering that matrilins have been found to play role in cell growth and tissue remodeling, their possible involvement in carcinogenesis has been raised. The main objective of this study was to investigate the changes in matrilin-2 expression which is one of the main components of basement membranes. Thirty-five cases of surgically resected hepatocellular carcinomas, 35 corresponding surrounding liver tissues and 10 normal liver samples were used for the study. In 15 of 35 cases the tumor developed on the basis of cirrhosis. Matrilin-2 protein expression was detected in normal liver around bile ducts, portal blood vessels, while sinusoids were negative by immunohistochemistry. Cirrhotic surrounding tissue showed intensive matrilin-2 staining along the sinusoids. Tumorous neovasculature was found strongly positive by immunohistochemistry. No differences, however, were detected by morphometry regarding the amount of protein expression based on the grade of hepatocellular carcinomas. Real-time RT-PCR did not show significant differences in matrilin-2 mRNA expression between normal, cirrhotic and tumor samples. This suggests posttranslational modification of matrilin-2 manifesting in altered distribution in liver fibrosis. Our data indicate that matrilin-2 is a novel basement membrane component in the liver, which is synthetised during sinusoidal "capillarization" in cirrhosis and in hepatocellular carcinoma. This is the first report to describe the expression and distribution of matrilin-2 in human normal and cirrhotic liver as well as in hepatocellular carcinoma.     

Epithelial-Mesenchymal Transition Markers in HCVAssociated Hepatocellular Carcinoma: A Multivariate Follow Up Study

Objective: Validated markers to predict recurrence after surgical resection of hepatocellular carcinoma (HCC) are needed. Little data is available regarding epithelial-mesenchymal transition (EMT) markers in HCC. The objective of this study was to investigate the expression of EMT markers and their correlation with clinicopathological variables and survival in hepatitis C virus (HCV)-associated HCC. Methods: This longitudinal study included 109 cases of HCV-associated HCC treated with surgical resection. Nine different EMT markers (vimentin, E-cadherin, N-cadherin, Stat3, Snail1, Slug, Twist1, Zeb1 and integrin α5) were evaluated on liver tissue from HCC cases. Twenty fresh HCC samples from the studied cases were used for gene expression of EMT markers by quantitative real time polymerase chain reaction (PCR). Results: EMT markers expression was 71%, 25%, 26%, 27%, 9%, 4%, 72%, 47%, 87% for vimentin, E-cadherin, N-cadherin, Stat3 snail1, slug, twist1, Zeb1 and integrin α5 respectively.  EMT mRNA in HCC tissues correlated with protein expression by 50-70%.  Vimentin was independent predictor of large tumor size (P=0.001), high risk of recurrence (HRR) (P=0.006) and shorter disease free survival (P=0.03) in multivariate analysis.  Reduced E-cadherin was a predictor of HRR (P=0.002). Conclusion: Vimentin and E-cadherin were the most powerful prognostic EMT markers in HCV-associated HCC in prediction of recurrence.     

Potential of Apolipoprotein A1 (ApoA1) for Detecting Liver Cirrhosis and Hepatocellular Carcinoma

Objective: Liver cirrhosis and hepatocellular carcinoma (HCC) are chronic liver diseases that can cause serious health problems. Meanwhile, the methods used to detect liver cirrhosis and HCC are limited. Apolipoprotein A1 (ApoA1) is a protein that makes up high-density lipoprotein (HDL), which plays a role in liver cirrhosis and HCC, and can be used as a biomarker. This study aims to determine the ability of ApoA1 to detect and differentiate liver cirrhosis and hepatocellular carcinoma. Methods: This cross-sectional study was conducted on 47 patients with liver cirrhosis and HCC at Margono Soekarjo Regional General Hospital, Purwokerto, Indonesia. This study also involved 33 healthy participants from blood donors at the Blood Transfusion Unit, Indonesian Red Cross, Banyumas. Serum ApoA1 levels were analyzed by ELISA method. Receiver Operating Characteristics (ROC) were used to evaluate the diagnostic power of ApoA1 and differentiate between cirrhotic, HCC, and healthy patients. Multivariate binary logistic regression test to determine the most influential variables on the incidence of cirrhosis, HCC, and health. Results: ApoA1 was able to differentiate cirrhosis from HCC, cirrhosis from healthy and HCC from healthy, with sensitivity 56.7%, 86.7%, 70.6%, specificity 70.6%, 93.9%, 84.9%, respectively, and AUC 68.5%, 92.6%, 75.0%. AFP (p = 0.002, OR 1.004) and bilirubin (p = 0.021, OR 1.259) were variables that contributed to cirrhosis - HCC. Age (p = 0.011, OR 0.766) and AST (p = 0.003, OR 0.834) are variables that play a role in health - cirrhosis. ALT (p = 0.024, OR 0.965) and PT (p = 0.004, OR 0.253) are variables that play a role in healthy - HCC. Conclusion: ApoA1 was best for detecting healthy from cirrhosis, followed by healthy from HCC and cirrhosis from HCC. ApoA1 is not the primary variable determining the incidence of cirrhosis - HCC, healthy - HCC, and healthy – HCC.     

肝细胞癌中NY-SAR-35 NY-TLU-57和NY-ESO-1基因的表达及意义

目的:检测三种癌-睾丸抗原NY-SAR-35、NY-TLU-57和NY-ESO-1基因mRNA在广西地区肝细胞癌(Hepatocellularcarcinoma,HCC)中的表达,探讨其作为HCC特异性免疫治疗靶抗原的可能性。方法:利用逆转录-聚合酶链反应(RT-PCR)方法,检测NY-SAR-35、NY-TLU-57和NY-ESO-1基因在63例HCC组织、56例HCC癌旁组织和4例正常肝组织中的表达,随机选择4例RT-PCR阳性产物直接进行DNA序列测定,并将其表达结果与临床病理指标进行统计学分析。结果:所检测的三种基因在4例正常肝组织中无表达;在63例HCC组织中,NY-SAR-35表达率为38.1%(24/63),NY-TLU-57为4.8%(3/63),NY-ESO-1为23.8%(15/63);在56例HCC癌旁组织中,NY-TLU-57和NY-ESO-1无表达,NY-SAR-35的表达率为26.8%(15/56)。基因表达与临床病理指标关系的分析显示,这三种癌-睾丸抗原的表达均与所分析的临床病理指标无关(P>0.05)。结论:癌-睾丸抗原NY-SAR-35、NY-TLU-57和NY-ESO-1基因可特异性的表达于HCC中,其中NY-SAR-35、NY-ESO-1具有一定的表达频率,提示它们有可能作为HCC特异性免疫治疗的靶抗原。     

Detection of MAGE-4 Protein in the Sera of Patients with Hepatitis-C Virus-associated Hepatocellular Carcinoma and Liver Cirrhosis

The aim of this study was to determine whether MAGE-4 protein is detectable in sera of patients with hepatocellular carcinoma and other liver diseases. An enzyme-linked immunosorbent assay was employed for detection of MAGE-4 protein in sera of liver disease patients, healthy men and women (control I) and those undergoing prostatic cancer screening (control II). MAGE-4 protein levels in sera of patients with hepatitis C virus-associated HCC (HCC-C) ( n =45, mean=2.160 ng/ml) and HCV-associated cirrhosis (LC-C) ( n = 55, 1.072 ng/ml) were significantly higher ( P < 0.0001) than those of control I (0.327 ng/ml) or control II (0.394 ng/ml). MAGE-4 protein was positive in 21/45 (46.7%) HCC-C patients and 18/55 (32.7%) LC-C patients (cut-off, mean plus 2 SD in healthy controls) but in 0/12 (0%) hepatitis B virus-associated HCC (HCC-B) patients, 3/49 (6.1%) hepatitis B virus-associated LC (LC-B) patients, 4/47 (8.5%) alcoholic liver disease patients, and 1/49 (2.0%) controls. Serum MAGE-4 protein level may be useful as a marker for identification of LC-C patients suffering from HCC that is undetectable by presently available methods.     

肝癌及肝硬化患者可溶性endoglin的临床意义*

目的：本研究拟探讨肝癌及肝硬化患者血清中可溶性endoglin 的表达及其临床意义。方法：采用酶联免疫吸附实验（ELISA）方法检测87例肝癌患者、30例肝硬化患者及28例健康对照组血清endoglin 的浓度,分析肝癌患者血清endoglin 表达与肝癌分期、门脉癌栓及远处转移等临床指标的关系。结果：肝癌合并肝硬化患者血清endoglin 浓度显著高于肝硬化组（P<0.001）及正常对照组（P<0.001）,肝硬化患者血清endoglin 浓度显著高于正常对照组（P=0.016）。 进一步研究发现,肿瘤最大径>5cm,有远处转移,有门脉癌栓、AFP>400ng/mL、肝癌临床分期Ⅲ期的患者血清endoglin 的浓度均明显升高（P<0.05）,ROC 分析发现endoglin和AFP 联合检测后敏感性提高,曲线下面积较前增加。结论：我们在国内首次阐明可溶性endoglin 在肝癌及肝硬化患者中的临床意义,血清endoglin 可作为评估乙肝肝硬化进展为肝癌的补充诊断指标,可能有一定早期诊断、预测复发转移及预测预后的价值。     

Comparison of Overall Survival between Transarterial Chemoembolization and Best Supportive Care in Intermediate- Stage Hepatocellular Carcinoma

Objective: The Thailand management guideline allows the use of transarterial chemoembolization (TACE) for the treatment of intermediate-stage hepatocellular carcinoma (HCC) in patients with decompensated cirrhosis, whereas other guidelines do not. The aim of this study was to compare the overall survival between TACE and the best supportive care (BSC) in HCC patients with Child–Pugh score 5–8 cirrhosis and in subgroups with compensated cirrhosis (Child–Pugh score 5–6) and early decompensated cirrhosis (Child–Pugh score 7–8). Methods: This retrospective study comprised 118 patients with intermediate-stage HCC. The overall survival was compared between TACE and BSC using the Kaplan–Meier method. Results: The median overall survival time for all patients was 21.4 months in the TACE group and 8.2 months in the BSC group (P <0.001). In the subgroup analyses, the overall survival times for TACE and BSC were 26 months and 9 months, respectively, for compensated cirrhosis (P <0.001), and 14.5 months and 6.9 months, respectively, for early decompensated cirrhosis (P <0.001). In the Cox proportional-hazards model, TACE was an independent prognostic factor for prolonged overall survival in all patients [hazard ratio (HR) 0.29; 95% confidence interval (CI), 0.17–0.49; P <0.001], patients with compensated cirrhosis (HR, 0.31; 95% CI, 0.16–0.62; P <0.001), and patients with early decompensated cirrhosis (HR, 0.16; 95% CI, 0.061–0.44; P <0.001). Conclusion: TACE improves the overall survival in patients with intermediate-stage HCC and compensated or early decompensated cirrhosis.