N‐glycan based models improve diagnostic efficacies in hepatitis B virus‐related hepatocellular carcinoma

The early diagnosis of hepatocellular carcinoma (HCC) is of great clinical desirable due to lack of specific and sensitive markers. Alterations in the sugar chains of glycoprotein synthesized by the liver contribute to the molecular basis of abnormalities in carcinogenesis. This study aims to construct and assess the diagnostic value of N‐glycan based diagnostic model in HCC identification and follow‐up. A total of 393 subjects including HBV‐related HCC, liver fibrosis and healthy controls were recruited. Follow‐up was carried out before and after surgical treatment in HCC. N‐glycome of serum glycoprotein was profiled by DNA sequencer‐assisted fluorophore‐assisted carbohydrate electrophoresis (DSA‐FACE). Multiparameters diagnostic models were constructed based on N‐glycan markers. The result found that 2 N‐glycan structure abundances (NG1A2F, Peak 4; NA3Fb, Peak 9) were useful as N‐glycan markers. The diagnostic efficacy of the log ratio [log(p9/4)] was similar to that of AFP in differentiating HCC from fibrosis. The accuracy and sensitivity of the diagnostic model combining AFP and N‐glycan markers (Cscore B) were increased 7–10% compared with that of AFP. Log(p9/4) was more efficient in monitoring the progression of HCC with regarding to vascular invasion at improved specificity (16%) and accuracy (8%) compared with that of AFP. The N‐glycan markers were found to be changed significantly after surgical resection in HCC follow‐up. We conclude that the branching α (1,3)‐fucosylated triantennary glycan and a biantennary glycan are promising as N‐glycan markers. The diagnostic models based on the N‐glycan markers and AFP improve the efficacy in HCC diagnosis and progression monitoring.     

Liver‐intestine cadherin predicts microvascular invasion and poor prognosis of hepatitis B virus‐positive hepatocellular carcinoma

BACKGROUND:

Liver‐intestine cadherin (LI‐cadherin; CDH‐17) is a new member of the cadherin superfamily with distinct structural and functional features. The study was designed to investigate the role of LI‐cadherin in tumor invasion and prognosis of human hepatitis B virus (HBV)‐positive hepatocellular carcinoma (HCC).

METHODS:

LI‐cadherin expression in HBV‐positive hepatocellular carcinoma cell lines with low‐ and high‐invasive potentials was evaluated by Western‐blot, immunofluorescence, and real‐time polymerase chain reaction (PCR) analyses. The role of LI‐cadherin in tumor invasion was also evaluated in vitro by a small‐interfering ribonucleic acid (siRNA)‐mediated approach. The prognostic significance of LI‐cadherin was validated in a cohort of HBV‐positive HCC patients by immunohistochemistry and Western‐blot.

RESULTS:

Significant high levels of LI‐cadherin mRNA and protein were found in the high‐invasive HCCLM3 as compared with those in low‐invasive PLC/PRF/5 and Hep3B cell line. Cell migration, adhesion to extracellular matrix, and matrigel invasion were significantly reduced after LI‐cadherin knockdown in HCCLM3 cells. Immunohistochemical analysis of 255 HBV‐positive HCC cases showed that overexpression of LI‐cadherin was well correlated with microvascular invasion, which was confirmed by Western‐blot in 32 tumor tissues, and its overexpression was strongly associated with shorter overall survival as well as higher incidence of tumor recurrence.

CONCLUSIONS:

LI‐cadherin is predictive of microvascular invasion and poor prognosis of HBV‐positive HCC, and would be a potential useful intervention target for HCC. Cancer 2009. © 2009 American Cancer Society.     

乙型肝炎病毒感染与原发性肝癌相关的危险度分析

目的:分析原发性肝癌(PHC)患者血清中乙型肝炎病毒(HBV)标志情况,探讨HBV感染与PHC危险性的关系。方法:收集在中国医科大学附属第一医院介入科住院的PHC患者224例作为病例组,同时期在此科住院的其他非肝病患者280例作为对照组,记录其血清HBV标志,与既往乙型肝炎和肝硬化等情况进行病例对照研究。结果:PHC组HBV总感染率为68.22%(146/214),以HBsAg阳性(65.42%,140/214)和抗-HBe阳性(30.84%,66/214)为主,对照组总感染率为7.63%(20/262),其中以HBsAg(3.44%,9/262)和抗-HBe(7.63%,20/262)为主。病例组感染率明显高于对照组(χ2=200.1,P<0.000 1),优势比OR=27.4(95%CI为16.0~46.9)。结论:HBV慢性感染是PHC的一个危险因素,在HBV慢性感染者中进行PHC筛查,对早期诊断PHC提高其临床疗效有重要意义。     

Serum interleukin‐6 associated with hepatocellular carcinoma risk: A nested case–control study

Inflammatory markers have been associated with increased risk of several cancers, including colon, lung, breast and liver, but the evidence is inconsistent. We conducted a nested case–control study in the longitudinal cohort of atomic‐bomb survivors. The study included 224 hepatocellular carcinoma (HCC) cases and 644 controls individually matched to cases on gender, age, city and time and method of serum storage, and countermatched on radiation dose. We measured C‐reactive protein (CRP) and interleukin (IL)‐6 using stored sera obtained within 6 years before HCC diagnosis from 188 HCC cases and 605 controls with adequate volumes of donated blood. Analyses with adjustment for hepatitis virus infection, alcohol consumption, smoking habit, body mass index (BMI) and radiation dose showed that relative risk (RR) of HCC [95% confidence interval (CI)] in the highest tertile of CRP levels was 1.94 (0.72–5.51) compared to the lowest tertile (p = 0.20). RR of HCC (95% CI) in the highest tertile of IL‐6 levels was 5.12 (1.54–20.1) compared to the lowest tertile (p = 0.007). Among subjects with BMI > 25.0 kg/m², a stronger association was found between a 1‐standard deviation (SD) increase in log IL‐6 and HCC risk compared to subjects in the middle quintile of BMI (21.3–22.9 kg/m²), resulting in adjusted RR (95% CI) of 3.09 (1.78–5.81; p = 0.015). The results indicate that higher serum levels of IL‐6 are associated with increased HCC risk, independently of hepatitis virus infection, lifestyle‐related factors and radiation exposure. The association is especially pronounced among subjects with obesity.     

Serum IL‐6 levels and the risk for hepatocarcinogenesis in chronic hepatitis C patients: An analysis based on gender differences

Interleukin‐6 (IL‐6) may play a role in the pathogenesis of hepatocellular carcinoma (HCC). Recently, it was reported in mouse models that estrogen‐mediated inhibition of IL‐6 production explains the gender disparity in HCC. We conducted a retrospective cohort study to examine whether this hypothesis is applicable to human HCC. We enrolled 330 patients with chronic hepatitis C whose serum samples were collected between January 1994 and December 2002. Serum IL‐6 concentrations were measured and patients were divided into three groups according to IL‐6 levels: low, middle, and high. We evaluated the association between serum IL‐6 levels and the risk of subsequent HCC development, including subgroup analysis on each gender. During the follow‐up period (mean 9.0 yr), HCC developed in 126 patients. The incidence rates differed significantly among the three groups (p = 0.015), increasing in accordance with serum IL‐6 levels. However, unexpectedly, this tendency was significant only in female patients. In a multivariate analysis, higher serum IL‐6 level was an independent risk factor for HCC development in female patients, with a hazard ratio of 1.61. Although female patients showed a weak negative correlation between serum IL‐6 levels and estradiol levels, the lower risk of HCC in female patients cannot be fully explained by estrogen‐mediated inhibition of IL‐6 production. In conclusion, higher serum IL‐6 level was an independent risk factor for HCC development in female but not male chronic hepatitis C patients. Measurement of serum IL‐6 levels may provide useful information for predicting future HCC development in female chronic hepatitis C patients. © 2009 UICC     

Interleukin-6 predicts recurrence and survival among head and neck cancer patients

BACKGROUND: Increased pretreatment serum interleukin (IL)-6 levels among patients with head and neck squamous cell carcinoma (HNSCC) have been shown to correlate with poor prognosis, but sample sizes in prior studies have been small and thus unable to control for other known prognostic variables. METHODS: A longitudinal, prospective cohort study determined the correlation between pretreatment serum IL-6 levels, and tumor recurrence and all-cause survival in a large population (N = 444) of previously untreated HNSCC patients. Control variables included age, sex, smoking, cancer site and stage, and comorbidities. Kaplan-Meier plots and univariate and multivariate Cox proportional hazards models were used to study the association between IL-6 levels, control variables, and time to recurrence and survival. RESULTS: The median serum IL-6 level was 13 pg/mL (range, 0-453). The 2-year recurrence rate was 35.2% (standard error, 2.67%). The 2-year death rate was 26.5% (standard error, 2.26%). Multivariate analyses showed that serum IL-6 levels independently predicted recurrence at significant levels [hazard ratio (HR) = 1.32; 95% confidence interval (CI), 1.11 to 1.58; P = .002] as did cancer site (oral/sinus). Serum IL-6 level was also a significant independent predictor of poor survival (HR = 1.22; 95% CI, 1.02 to 1.46; P = .03), as were older age, smoking, cancer site (oral/sinus), higher cancer stage, and comorbidities. CONCLUSIONS: Pretreatment serum IL-6 could be a valuable biomarker for predicting recurrence and overall survival among HNSCC patients. Using IL-6 as a biomarker for recurrence and survival may allow for earlier identification and treatment of disease relapse.     

Skull metastasis from hepatocellular carcinoma with chronic hepatitis B

A 56-year-old male visited our hospital for evaluation of an occipital mass. Contrast computed tomography showed hypervascular enhancement with osteolytic change in the skull and a huge enhanced mass in the liver. Magnetic resonance imaging showed bone metastasis in the thoracic vertebrae. Assays for hepatitis B surface antigen and hepatitis B core antibody were positive and his liver condition was Child-Pugh grade A. Our diagnosis was hepatocellular carcinoma (HCC) with skull and vertebrae metastases on chronic hepatitis B. He was treated with radiation therapy for bone metastases and transcatheter arterial chemoembolization for HCC. But he developed acute respiratory failure because of aspiration pneumonia, congestion and oedema with haemorrhage of the lungs and died. Dissection showed HCC with multiple bone metastases. The liver tumor was categorized as well-differentiated HCC, Edmondson classification I, trabecular type and pseudoglandular type. In the liver mild infiltration of lymphocytes was seen in Glisson's capsules which were significantly enlarged with well preserved limiting plates. Piecemeal necrosis was not obvious. No fibrosis was noted. An 8 cm × 7 cm × 3 cm metastatic lesion had formed in the left occipitotemporal part of the cranial bone. The lesion was osteolytic and showed invasion into the dura mater. Neither the subdural cavity nor the brain showed involvement from the metastatic tumor. However, skull metastasis from HCC is very rare and it affects the patient's prognosis and the quality of life. Therefore, it is very important to make an early diagnosis and carry out proper management of skull metastasis from HCC.     

A meta-analysis of case-control studies on the combined effect of hepatitis B and C virus infections in causing hepatocellular carcinoma in China

We investigated whether concurrent infection by hepatitis B virus (HBV) and hepatitis C virus (HCV) in China, a hyperepidemic area for these infections, was associated with a higher risk of causing hepatocellular carcinoma (HCC) than each infection alone in a meta-analysis in China, 32 case-control studies involving 3201 cases and 4005 controls, identified from a computer-based literature search from 1966 to 2004. The pooled odds ratio and 95% confidence interval (CI) for HBsAg positivity was 14.1 (95% CI: 10.6-18.8); for anti-HCV/HCV RNA positivity was 4.6 (95% CI: 3.6-5.9); for HBsAg positivity and anti-HCV/HCV RNA negativity were 15.6 (95% CI: 11.5-21.3); for HBsAg negativity and anti-HCV/HCV RNA positivity were 8.1 (95% CI: 5.0-13.0); and positivity for both HBsAg and anti-HCV/HCV RNA was 35.7 (95% CI: 26.2-48.5). We conclude that HBV and HCV infections are important independent risk factors for HCC in China, and that dual infection by HBV and HCV is associated with a higher risk of causing HCC than each infection alone, suggesting a synergism between HBV and HCV.     

Association of variants in the interleukin‐27 and interleukin‐12 gene with nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) is multifactorial, and the genetic background may be a crucial etiologic factor. Interleukin‐12 (IL‐12) is a multifunctional cytokine that induces interferon (IFN)‐gamma secretion and plays an important role in antitumor immunity. Interleukin‐27 (IL‐27) is a novel IL‐12 family member, the present studies demonstrate that IL‐27 mediates potent antitumor activity. Variations in the DNA sequence in the IL‐12 and IL‐27 gene may lead to altered cytokines production and/or activity, and so this can modulate an individual's susceptibility to NPC. To test this hypothesis, we investigated the relationship of IL‐12 and IL‐27 gene polymorphisms and NPC in a Chinese population. We analyzed single nucleotide polymorphisms (SNPs) of IL‐12 gene 16974 A/C and IL‐27 gene ?964 A/G, 2905 T/G, 4730 T/C in 302 patients with NPC and 310 age‐ and sex‐matched controls, using polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) and DNA sequencing methods. There were significant differences in the genotype and allele distribution of 16974 A/C polymorphism of the IL‐12 gene among cases and controls. The 16974 CC and AC genotypes were associated with a significantly increased risk of NPC as compared with the 16974 AA genotypes (OR = 2.225, 95% CI 1.395–3.549, P = 0.001 and OR = 1.834, 95% CI 1.239–2.716, P = 0.002, respectively). The 16974 C allele was associated with a significantly increased risk of NPC as compared with the 16974 A allele (OR = 1.334, 95% CI 1.065–1.670, P = 0.012). However, genotype and allele frequencies of the IL‐27 gene ?964 A/G, 2905 T/G, 4730 T/C polymorphisms in NPC patients were not significantly different than that in healthy controls (P > 0.05). Our data suggest that IL‐12 gene may play a role in the development of NPC. © 2009 Wiley‐Liss, Inc.     

慢性乙型肝炎e系统状态与肝纤维化及癌变的关系

目的：为了弄清慢性乙型肝炎病程中乙型肝炎病毒ｅ系统状态和纤维化进展及其癌变的关系。方法：分别比较慢性乙型肝炎不同纤维化分期以及原发性肝癌组的ＨＢｅＡｇ或抗－ＨＢｅ或抗性率。结果：ＨＢｅＡｇ阳性率在慢性乙型肝炎纤维化Ｓ１、Ｓ２、Ｓ３和Ｓ４分期中分别为８５．２％（２３／２７）、７２．７％（２４／３３）、６３．３％（１９／３０）和５０％（１２／２４），原发性肝癌组为１９．４％（６／３１）；Ｓ１和Ｓ４组比较，以及原发性肝癌组和Ｓ１、Ｓ２、Ｓ３且比较存在统计学差异（Ｐ〈０．００５－０．０１），抗－ＨＢｅ阳性率在Ｓ１、Ｓ２、Ｓ３和Ｓ４分期中分别为１１．１％（３／２７）、２１．２％（７／３３）、２６．７％（８／３０）和３７．５％（９／２４），原发性肝癌组为６１．３％（１９／３１）；Ｓ１和Ｓ４组比较，以及原发性肝癌组和Ｓ１、Ｓ     

Type 2 diabetes and hepatocellular carcinoma: a case-control study in patients with chronic hepatitis B

Type 2 diabetes has been suggested as an independent risk factor for the development of hepatocellular carcinoma (HCC). However, the role of Type 2 diabetes on the development of HCC in the presence of chronic hepatitis B (CHB) remains inconclusive. We conducted this hospital-based case-control study to evaluate the roles of Type 2 diabetes in HCC development in patients with CHB. From January 2004 to December 2008, a total of 6,275 eligible consecutive patients with chronic hepatitis B virus (HBV) infection were recruited. A total of 1,105 of them were patients with HBV-related HCC and 5,170 patients were CHB but without HCC. We used multivariate logistic regression models to investigate the association between Type 2 diabetes and HCC risk. The prevalence of Type 2 diabetes is higher among HCC patients without cirrhosis than among those with cirrhosis (12.1% vs. 6.7%, p=0.003). Type 2 diabetes was associated with a significantly high risk of HCC in female patients after adjusting for age, family history of HCC, city of residence, hepatitis B e antigen and cirrhosis with an odds ratio (95% confidence interval, CI) of 1.9 (1.1-3.4). Restricted analyses among female patients without cirrhosis indicated that Type 2 diabetes was strongly associated with HCC risk with adjusted odds ratio (95% CI) of 5.6 (2.2-14.1). In conclusion, Type 2 diabetes is independently associated with the increased risk of HCC in female CHB patients. Female CHB patients with Type 2 diabetes are of a high HCC risk population and should be considered for HCC close surveillance program.     

慢性乙型肝炎病人血清COX-2表达的临床意义

目的为了探求慢性乙型肝炎患者血清中环氧化酶-2（COX-2）的表达情况及其临床意义。方法选择正常对照组10例,慢性乙型肝炎轻、中、重度各10例,肝硬化病人10例作为本实验的研究对象,应用ELISA法进行血清样本COX-2测定,并结合病情轻重进行综合分析。结果慢性乙型肝炎组和肝硬化组血清中COX-2阳性率明显高于对照组（P〈0．01）,血清COX-2含量显著高于对照组（P〈0．01）。慢性乙型肝炎各组病人组血清中COX-2含量存在显著的差异（P〈0．05）。结论COX-2在慢性乙型肝炎患者血清中呈现出高表达,并且在其发病过程中起着重要的作用。     

Aflatoxin B1 exposure increases the risk of cirrhosis and hepatocellular carcinoma in chronic hepatitis B virus carriers

The relation between aflatoxin B₁ (AFB₁) and cirrhosis in chronic carriers of hepatitis B virus (HBV) remains inconclusive. This case‐control study nested in a large community‐based cohort aimed to assess the effect of AFB₁ exposure on cirrhosis and HCC in chronic HBV carriers. Serum AFB₁‐albumin adduct levels at study entry were measured in 232 cirrhosis cases, 262 HCC cases and 577 controls. Multivariate‐adjusted odds ratios (aORs) and 95% confidence intervals (95% CIs) were estimated using logistic regression. Among all chronic HBV carriers, the time intervals between study entry and diagnosis of HCC, cirrhosis, cirrhotic HCC, and non‐cirrhotic HCC were all significantly (p < 0.0001) shorter in participants with high serum levels of AFB₁‐albumin adducts than those with low/undetectable levels. There were significant dose‐response relations with serum AFB₁‐albumin adduct level at study entry for cirrhosis (p‐trend = 0.0001) and cirrhotic HCC (p‐trend < 0.0001) newly diagnosed within 9 years after entry as well as non‐cirrhotic HCC (p‐trend = 0.021) newly diagnosed within 4 years after entry. The aORs (95% CIs) for high versus undetectable serum AFB₁‐albumin adduct levels were 2.45 (1.51–3.98) for cirrhosis (p = 0.0003), 5.47 (2.20–13.63) for cirrhotic HCC (p = 0.0003), and 5.39 (1.11–26.18) for non‐cirrhotic (p = 0.0368) HCC, respectively. There remained a significant dose‐response relation between serum AFB₁‐albumin adduct level and HCC risk (p‐trend = 0.0291) in cirrhosis patients, showing an aOR (95% CI) of 3.04 (1.11–8.30) for high versus undetectable serum levels (p = 0.0299). It is concluded that AFB₁ exposure may increase the risk of cirrhosis and HCC in a dose‐response manner among chronic HBV carriers.