Serum Markers of Hepatocellular Carcinoma

Background   

The hepatocellular carcinoma is one of the most common malignant tumors and carries a poor survival rate. The management of patients at risk for developing HCC remains intricate.     

Serum Alpha-Fetoprotein Has High Specificity for the Early Detection of Hepatocellular Carcinoma After Hepatitis C Virus Eradication in Patients

Alpha-fetoprotein (AFP) has not played a large role in the surveillance of hepatocellular carcinoma due to inadequate sensitivity and specificity for active chronic hepatitis or cirrhosis. The aim of this study was to evaluate the diagnostic accuracy of AFP in small hepatocellular carcinomas after hepatitis C virus eradication to determine the optimal cutoff value.We conducted a case–control study of 29 cases and 58 controls, matched for age, gender, and platelet counts.The AFP cutoff was 5 ng/mL in patients after hepatitis C virus eradication and 17 ng/mL in those without hepatitis C virus eradication. The areas under the receiver operating characteristic curve were 0.86 (95% confidence interval, 0.76–0.96) in patients after hepatitis C virus eradication and 0.83 (95% confidence interval, 0.74–0.91) in those without hepatitis C virus eradication. In patients after hepatitis C virus eradication, the sensitivity and specificity of AFP levels were 24.1% and 100%, respectively, using a cutoff value of 17 ng/mL. Using a lower cutoff value of 5 ng/mL, the sensitivity increased to 75.9%, although the specificity decreased to 89.0%.AFP is a specific tumor marker for the diagnosis of hepatocellular carcinoma after hepatitis C virus eradication when using the optimal cutoff value of 5 ng/mL.     

Detection of Epstein-Barr Virus DNA in Hepatocellular Carcinoma Tissues from Hepatitis C-positive Patients

Background: We have previously shown that hepatitis C virus (HCV) replication is promoted by the Epstein-Barr virus (EBV) in vitro. The aim of this study was to examine the EBV load in hepatocellular carcinoma (HCC) tissues from HCV antibody-positive patients. Methods: DNA was extracted from paraffin sections from 168 HCC patients. After amplification of a region in the EBV BamHI W sequence by means of the polymerase chain reaction (PCR), it was detected by Southern hybridization and semi-quantified. Ten hyperplastic lesions from HCV-positive patients and 35 non-tumorous samples from hepatitis-negative patients served as controls. The PCR results were analyzed on the basis of the patient's hepatitis status. Univariate and multivariate analyses were performed to identify clinicopathologic factors for predicting EBV infection in HCC tissues. Results: More than one copy of EBV DNA per 100 cells was detected in 56 (33%) of the HCC sections. The detection ratio in HCC tissues from HCV antibody-positive patients was 40% (45 of 113), which was significantly higher than that in tissues from HBV surface antigen-positive patients (14%, 5 of 37; P = 0.0018). The patient's serum HBV surface antigen and HCV antibody independently predicted the EBV positivity of HCC tissues. Conclusions: These results support our hypothesis that EBV could play an important role in the development of HCV-related HCC.     

胃癌患者血清RASSF1A基因启动子区甲基化检测

Ras相关结构域家族1A（RASSF1A）基因是近年发现的新型抑癌基因，其启动子区甲基化可能与胃肠道肿瘤的发生、发展密切相关。目的：检测胃癌患者血清RASSF1A基因启动子区甲基化情况，探讨其在胃癌早期诊断和预后评估中的可能作用。方法：以甲基化特异性聚合酶链反应（MSP）检测47例胃腺癌患者、30例胃良性病变患者和30名健康对照者的血清RASSF1A基因启动子区甲基化情况，其中16例胃腺癌患者同时留取手术切除癌组织、癌旁组织标本以及术前、术后血标本行对照研究。结果：胃腺癌患者血清RASSF1A基因启动子区甲基化率为34．0％（16／47），显著高于胃良性病变患者（3.3％，1／30）和健康对照者（0％）（P〈0．01）。16例胃腺癌组织中5例（31．2％）检测到RASSF1A基因启动子区甲基化，其中4例（80．0％）术前、术后血清标本均检测到RASSF1A基因启动子区甲基化。血清RASSF1A基因启动子区甲基化与胃癌患者性别、年龄、肿瘤分化程度、有无转移以及血清癌胚抗原（CEA）水平均无相关性。结论：血清RASSF1A基因启动子区甲基化检测可望为胃癌的早期诊断和预后判断提供依据。     

Hypercoagulable States in Patients with Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) patients have an increased risk for venous thromboembolism, mainly portal venous thrombosis (PVT). The aim of this study was to assess the role of acquired and hereditary thrombotic risk factors in HCC patients. Thirty-one patients with HCC, 30 patients with cirrhosis but without HCC or PVT, and 48 matched healthy controls were studied. Mean levels of plasma protein C, protein S, antithrombin, and serum lipoprotein (a) were significantly lower in patients with HCC and in the cirrhotic group compared to the healthy controls. Mean serum homocysteine levels were significantly higher in patients with HCC compared to cirrhotics and healthy controls. The prevalence of activated protein C resistance, factor V Leiden mutation, prothrombin gene mutation G20210GA, and C677T methylenetetrahydrofolate reductase polymorphism was not significantly different among the three groups. In conclusion, thrombophilic defects are common in HCC patients and they might contribute to the observed thrombotic complications in this malignancy.     

The Influence of Alcoholic Liver Disease on Serum PIVKA-II Levels in Patients without Hepatocellular Carcinoma

Background/AimsProthrombin induced by vitamin K deficiency or antagonist II (PIVKA-II) is a widely used diagnostic marker for hepatocellular carcinoma (HCC). We evaluated the correlation between alcoholic liver disease (ALD) and serum PIVKA-II levels in chronic liver disease (CLD) patients.MethodsWe retrospectively reviewed the medical records of 2,528 CLD patients without HCC. Among these patients, 76 exhibited serum high PIVKA-II levels of >125 mAU/mL (group 1). We categorized 76 control patients matched by age, sex, and the presence of liver cirrhosis from the remaining patients who were negative for serum PIVKA-II (group 2).ResultsGroup 1 revealed increased antibiotic usage (23.7% vs 2.6%, p<0.001) and incidence of ALD (60.5% vs 14.5%, p<0.001) as well as elevated aspartate aminotransferase (52.5 IU/L vs 30.5 IU/L, p=0.025) and γ glutamyl transpeptidase (67.5 IU/L vs 36.5 IU/L, p=0.005) levels compared with group 2. Further, group 1 was significantly associated with a worse Child-Pugh class than group 2. In the multivariate analysis, ALD (odds ratio [OR], 7.151; p<0.001) and antibiotic usage (OR, 5.846; p<0.001) were significantly associated with positive PIVKA-II levels.ConclusionsOur study suggests that ALD and antibiotics usage may be confounding factors when interpreting high serum PIVKA-II levels in patients without HCC. Therefore, serum PIVKA-II levels in patients with ALD or in patients administered antibiotics should be interpreted with caution.     

原发性肝癌患者血清瘦素及血脂水平的研究

目的 探讨原发性肝癌患者血清瘦素水平及其异常变化的临床意义.方法 选择64例原发性肝癌患者和59例与其在性别、年龄和体重指数(BMI)相匹配的对照者,应用ELISA方法检测血清瘦素水平,同步检测肝功能和血脂,肝癌患者测定甲胎蛋白,对血清瘦素、血脂与肝功能的关系进行分析.结果 肝癌组与对照组相比血清瘦素水平无差异;与正常对照组相比,肝癌患者CHO、LDL水平显著降低,差异有显者性.去除腹水因素,无腹水肝癌患者血清瘦素水平与BMI及Fat%均呈显著正相关,与肝功能、甲胎蛋白无关.结论 血清瘦素水平不能作为原发性肝癌患者肝功能的预测指标,血脂是肝功能减退的指标.瘦素可能参与了肝癌患者的营养不良.     

Analysis of the Genome-Wide DNA Methylation Profile of Side Population Cells in Hepatocellular Carcinoma

Background

DNA methylation plays an important role in maintaining pluripotency and regulating the differentiation of stem cells, but the DNA methylation profile of stem cells in hepatocellular carcinoma (HCC) remains unclear.

Aims

To investigate the genome-wide DNA methylation profile of side population (SP) cells of HCC, a special subpopulation of cells enriched with cancer stem cells, by DNA methylation microarray analysis and to analyze the functions and signal pathways of the aberrantly methylated genes in SP cells.

Methods

Side population cells were isolated from HCC cell lines Huh7 and PLC/PRF/5 using flow cytometry, and the tumorigenicity of these SP cells was assessed in NOD/SCID mice. The genome-wide DNA methylation status of SP cells and non-SP (NSP) cells was detected and compared by DNA methylation microarray analysis. Genes with differential methylation between SP and NSP cells were further analyzed for their functions and roles in related signaling pathways.

Results

Subcutaneous inoculation of 1 × 10³ SP cells yielded tumors in 60 % NOD/SCID mice, whereas no tumor was developed after the inoculation of 1 × 10⁶ NSP cells. Genome-wide DNA methylation microarray analysis showed that 72 and 181 genes were hypermethylated and hypomethylated, respectively, in both Huh7 and PLC/PRF/5 SP cells as compared with their corresponding NSP cells. Analyses of signaling pathways revealed that hypermethylated and hypomethylated genes were related to four and eight pathways, respectively.

Conclusions

Hepatocellular carcinoma SP cells possessed a differential DNA methylation status compared with NSP cells, and the differentially methylated genes in SP cells were involved in 12 signaling pathways. Our results provide valuable clues for further investigations in elucidating the importance of epigenetic regulation in sustaining HCC SP cells and tumorigenesis.     

乙型肝炎病毒pX与Bcl-2在肝癌中的表达

目的:探讨肝癌组织中乙型肝炎病毒pX与BCl-2的表达状况及关系。方法:病理证实的34例肝癌组织经连续切片,常规脱蜡水化,用S-P法行pX与Bel-2免疫组织化学染色。结果:pX与Bcl-2在肝癌表达的阳性率分别为97.06％及17.65％。Redit分析示两组有显著差异(P<0.01),阳性信号均主要位于胞浆,且两者可在同一癌组织的相同区域出现阳性染色。结论:乙型肝炎病毒pX在肝癌组织中存在高强度的广泛表达,pX的表达可能具有激活Bcl-2抑制癌细胞凋亡的作用。     

肝癌细胞中γ-谷氨酰转肽酶基因甲基化研究

目的:研究肝癌细胞中γ-谷氨酰转肽酶(GGT)甲基化的改变。方法:纯化人肝癌及癌旁组织中总GGT及RNA,观察总GGT、膜结合和可溶性GGT的表达,并以逆转录巢式PCR扩增GGT非编码区基因片段,分析M_3位点的甲基化状态。结果:肝癌、癌旁和远癌组总RNA浓度呈明显的梯度升而趋势(P<0.05);癌组织总GGT、膜结合和可溶性GGT均高于癌周又远癌组织(P<0.05);在癌、癌旁及远癌组织中,特定基因片段的扩增检出率分别为100％、85％、75％,M_3位点低甲基化频率分别为75％、55％、50％。结论:肝癌组织中GGT呈异常表达与基因低甲基化状态有关。     

Hepatocellular Carcinoma in Patients with Acute Intermittent Porphyria

ABSTRACT In a retrospective study over a 20-year period we found in the Umeå region in Sweden 11 patients (7 women and 4 men, mean age 67 years) with both hepatocellular carcinoma and acute intermittent porphyria. This coincidence was highly significant. Concomitant existence of portal cirrhosis of the liver was demonstrated in those 5 patients in whom it could be examined.     

A Molecular Hepatocellular Carcinoma Prognostic Score System Precisely Predicts Overall Survival of Hepatocellular Carcinoma Patients

Background and Aims: With high rates of recurrence post-treatment, hepatocellular carcinoma (HCC) is one of the most common types of cancer worldwide and the ma...     

Clinical Significance of Serum Ferritin Determination for Hepatocellular Carcinoma

The clinical significance of serum ferritin as a serological marker of hepatocellular carcinoma (HCC) was studied. Fasting serum ferritin levels were measured in 343 patients with diseases of the liver, using a radioimmunoassay ferritin kit. Elevated ferritin levels were obtained in various liver diseases but hyperferritinemia could be more clearly interpreted by classifying ferritin levels according to serum iron or transaminase values. Significantly higher values were obtained in HCC than liver cirrhosis. Sensitivity for diagnosis of HCC increased by serial and simultaneous determinations of ferritin and alpha-fetoprotein because high ferritin levels were observed more often in low alpha-fetoprotein-producing HCC and also in HBsAg negative, alcohol related, small-sized HCC. Therefore, simultaneous determination of alpha-fetoprotein and ferritin seems to be useful for detection of HCC in high risk patients such as those with liver cirrhosis.     

Causes of Death in Patients with Unresectable Hepatocellular Carcinoma

Most patients with hepatocellular carcinoma (HCC) also have cirrhosis, an independent cause of death. We considered an alternative definition of tumor-related death in patients with HCC and attempted to validate our definition. Two hundred thirty-seven HCC patients were diagnosed, followed, and died over a 12-year period and were evaluated every 2 months, including their last 6 months of life. We defined death by cancer if there was, in the last 6 months of life, a CT scan increase of >25% in the sum of tumor index lesions’ cross-sectional areas or new onset of, or increase in, either vascular invasion or metastatic disease (Group 1). Patients with stable cancer were considered to have died from any other cause (Group 2). We found that 135 (57%) patients died from cancer progression (Group 1), whereas 102 (43%) patients did not (Group 2). There was a statistically significant difference between Group 1 and Group 2 patients in percentage with bilobar disease (P = 0.03), more than one tumor (P = 0.01), an increase in AFP (P = 0.04), vascular invasion (P = 0.001), and the presence of metastases (P = 0.01). We conclude that 57% of patients with unresectable HCC died as a direct result of cancer progression, but 43% did not. The latter died from complications of their cirrhosis, including sepsis, GI bleeds, and renal failure.