Isolation of ferritin from human hepatocellular carcinoma

Ferritin was extracted from human hepatocellular carcinoma tissue and purified using column chromatography, gradient gel electrophoresis and cadmium sulphate crystallization. DEAE cellulose chromatography showed a difference between hepatoma and normal liver ferritin, indicative of a more acidic isoferritin profile in the tumour. Column-purified and crystalline ferritin and that remaining in the mother-liquor after crystallization was subjected to isoelectric focusing. Hepatoma ferritin showed higher concentrations of acidic isoferritins than liver ferritin. This was most obvious with mother-liquor ferritin, as crystallization tended to select out more basic isoferritins. Subunit analysis of hepatoma and liver ferritin showed a higher proportion of heavy subunits in the tumour ferritin, in keeping with the presence of acidic isoferritins. An antibody against hepatoma mother-liquor ferritin was raised in rabbits. However, hepatoma ferritin proved to be antigenically identical with normal liver ferritin, and we were thus unable to develop a specific radioimmunoassay for hepatoma ferritin.     

Diagnostic value of serum ferritin in primary hepatocellular carcinoma

The diagnostic value of serum ferritin levels was evaluated in 19 patients with biopsy-proven primary hepatocellular carcinoma (PHC) and 26 patients with chronic liver disease (CLD). Serum ferritin levels were significantly elevated in PHC, as compared with CLD and controls (p less than 0.0005). Similarly, serum ferritin/SGOT ratio, an index of increased ferritin production, was significantly higher in PHC than in CLD and controls. Serum alpha-fetoprotein (alpha-FP) was higher in PHC than in CLD (p less than 0.0025). No significant correlation was noted between serum ferritin and alpha-fetoprotein or SGOT in PHC and CLD. 17 of 19 patients with PHC had serum ferritin values over 450 ng/ml (sensitivity 88%). By contrast, only 10 of 17 patients with PHC (59%) demonstrated alpha-FP levels over 25 ng/ml, compatible with the diagnosis of PHC. 9 of these 10 patients had ferritin levels over 450 ng/ml, within the distribution of values for PHC. Conversely, 7 of 17 patients with PHC (40%) had normal levels of alpha-FP (false-negative). However, 6 of these patients (86%) had ferritin levels over 450 ng/ml, consistent with values in PHC. In this study, the overall sensitivity of serum ferritin in PHC was higher than that of alpha-FP (88 versus 59%) and its specificity 85 versus 68% for alpha-FP. These data indicate that serum ferritin may be utilized as a useful diagnostic marker in the evaluation of patients with PHC.     

The clinical value of serum ferritin in hepatocellular carcinoma

Serum ferritin is often elevated in patients with hepatocellular carcinoma (HCC). Its use as a disease marker has been proposed. We have measured serum ferritin levels in 85 patients with HCC and in 62 comparable subjects with cirrhosis. Abnormal values (greater than or equal to 300 ng/ml) were found in 54% of the patients with HCC and in 35% of those with cirrhosis (median 323 and 204 ng/ml, respectively). The overlap of the range of concentration in HCC and cirrhosis was so great that no discriminant level could be chosen. No relationship was found between alpha-fetoprotein and ferritin concentrations. Among 61 patients who received Adriamycin treatment, no discernible fall in ferritin levels was observed, while alpha-fetoprotein increased progressively during the follow-up. Serum ferritin has no role in diagnosing and/or monitoring the response to treatment of patients with HCC.     

Serum markers of hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in some areas of the world with increasing incidence worldwide. Most of patients with HCC are diagnosed at a late stage. Therefore, the prognosis of HCC patients is generally very poor with a 5-year survival rate of less than 5%. Screening strategies including alpha-fetoprotein (AFP) and ultrasound every 6 months in patients with liver cirrhosis, the major risk factor for HCC development, have been recommended to detect HCC at earlier stages amenable to effective treatment strategies. AFP, however, is a marker with poor sensitivity and specificity and the ultrasound is highly dependent on the operator's experience. Apart from AFP, lens culinaris agglutinin-reactive AFP and des-gamma carboxyprothrombin and several other biomarkers (e.g., glypican-3, human hepatocyte growth factor, and insulin-like growth factor) have been proposed as markers for HCC detection. In addition, with recently employed techniques, such as gene-expressing microarrays and proteomics, it is to be expected that new HCC-specific markers will become available in the near future. For all such proposed markers, however, the clinical usefulness has to be carefully evaluated and validated.     

Serum N-acetylglucosaminyltransferase III activities in hepatocellular carcinoma

N-Acetylglucosaminyltransferase III (GnT III) catalyses the addition of N-acetylglucosamine through a β 1–4 linkage to the mannose of the trimannosyl core, resulting in conversion of the concanavalin A (Con A)-reactive glycan into a non-reactive state. In this study, we measured GnT III activity to evaluate its diagnostic efficacy and its therapeutic effect on hepatocellular carcinoma (HCC). Concanavalin A-non-reactive fraction of serum transferrin (Tf) was also determined since the sugar chains of Tf are one of the possible candidates for the product of GnT III. Serum samples (159) were used from patients with HCC (89), liver cirrhosis (30), chronic hepatitis (19), alpha-fetoprotein (AFP) producing gastric carcinoma metastatic to the liver (five) and healthy controls (16). N-Acetyl-glucosaminyltransferase III activity was determined by high performance liquid chromatography. The reactivity of serum Tf to Con A was also analysed in 21 paired HCC samples before and after treatment by crossed immuno-affinoelectrophoresis. N-Acetylglucosaminyltransferase III activity from the HCC group (153 ± 72 pmol/mL/h) was significantly higher than that from liver cirrhosis (99 ± 67 pmol/mL per h), chronic hepatitis (84 ± 39pmol/mL per h) and the normal controls (62 ± 16pmol/mL per h). N-Acetylgiucosaminyltransferase III activity of 21 patients with HCC was significantly reduced after treatment such as transcatheter arterial chemoembolization and/or percutaneous ethanol infection therapy, (123 ± 77 to 100 ± 60pmol/mL per h). Commensurate decreases of AFP and des-γ-carboxy prothrombin with GnT III activity were also observed after treatment. The Con A-non-reactive fraction (n= 21; 6.4 ± 2.3%) in patients with HCC after treatment was significantly lower than before (8.2 ± 2.4%). The present study suggests that GnT III activity is a possible and in the diagnosis and evaluation of HCC, especially when other tumour markers are negative.     

Serum ferritin

PURPOSE: There is increasing evidence that screening for colorectal cancer may save lives, and consequently, both professional and public interest in screening for colorectal cancer is increasing. As yet, however, there is no perfect screening test. Insidious blood loss is a common feature of colorectal cancer and may lead to a fall in serum ferritin before the patient becomes anemic. Measurement of serum ferritin, which is widely available and easily and inexpensively performed, has, therefore, been postulated as a potential screening test for colorectal cancer. METHOD: This study used samples of serum collected from 148 patients recruited to a screening study for colorectal cancer. All patients were thoroughly investigated by double-contrast barium enema and/or colonoscopy. Patients were selected randomly from each of three clinical diagnostic groups: 50 patients with proven colorectal cancer, 49 patients without colon disease, and patients with adenomas of the colon. Serum ferritin was assayed by immunoradiometry. The expected adult reference range is 25 to 350 µg/l, and results were reported without patient identification. RESULTS: There were no significant differences in serum ferritin levels among any of the three groups. CONCLUSION: Serum ferritin is unlikely to be of value as a screening test for colorectal cancer.     

Serum ferritin in beta-thalassaemia intermedia

The iron status of 10 patients with beta-thalassaemia intermedia has been investigated in order to assess whether the liver iron overload could be constantly predicted through the patients' S-ferritin levels. Our data have shown that only when the iron overload is in the far advanced stages do the S-ferritin concentrations appear to be strongly raised, whilst in the early stages of the disease the levels fall to normal ranges with consequent underestimation of the expanding iron stores. Thus, in the early stages of the iron overload the liver iron burden of such patients is only reflected by both chemical and histochemical investigations performed on the liver biopsy specimens long before the rise in their S-ferritin levels.     

Serum ferritin in hematologic malignancies

Serum ferritin was measured in a variety of hematologic malignancies at presentation, in remission following therapy, and in relapse. Ferritin was strikingly increased in all acute leukemias at presentation and in relapse, in the blastic crisis of CML, and in smouldering leukemia. Remission in both ALL and ANLL was associated with a reduction of serum ferritin, and this normalization was a function of remission duration. In the malignant lymphomas serum ferritin was related to tumor histology. Highest levels were found in Hodgkin disease and histiocytic lymphoma, normal levels in lymphocytic lymphoma, and intermediate levels in mixed histiocytic-lymphocytic lymphoma. In all cases, remission was associated with normalization of serum ferritin. These correlations suggest that serum ferritin measurements may be of clinical usefulness in the initial evaluation and in the assessment of response to therapy in patients with acute leukemia and malignant lymphoma.     

Serum ferritin in refractory anemias

The relationship between the level of erythropoiesis and iron balance was evaluated in 13 subjects with idiopathic refractory anemias. Serum ferritin levels and bone marrow iron stores were increased only in those patients with ring sideroblasts, erythroid hyperplasia and ineffective erythropoiesis. The magnitude of the increase correlated with the duration of anemia and the degree of increase in the erythron iron turnover. Ferritin levels were not related to the severity of the anemia, indicating that increased iron stores did not represent a shift of iron from the erythron or an absorption response to anemia per se. It does suggest that the level of erythroid proliferation directly affects gastrointestinal iron absorption, which in time leads to iron overload.     

Serum interleukin-8 levels in patients with hepatocellular carcinoma

Serum levels of interleukin-8 (IL-8) in 73 patients with hepatocellular carcinoma (HCC), 24 with liver cirrhosis (LC) and 18 with chronic hepatitis (CH), and in 20 healthy controls were measured by an enzyme-linked immunosorbent assay. The mean (± SD) level of serum IL-8 in patients with HCC was 48.4 ± 60.8 pg/ml, which were significantly (P < 0.01) higher than those in patients with LC (8.6 ± 8.6) and CH (2.7 ± 6.1), or that in controls (1.8 ± 5.8). It was revealed that serum IL-8 level was normalized after resection of HCC in patients whose serum IL-8 level had been high at the time of diagnosis. On the other hand, serum IL-8 levels were progressively increased with time in patients with HCC who had not received effective treatment. These findings suggest that IL-8 is produced by HCC cells in vivo, and serum IL-8 levels could be a marker for HCC. Moreover, significant correlation between serum IL-8 levels and the tumor sizes was found in patients with moderately differentiated HCC, which were mostly hypervascular on angiography. Serum IL-8 levels were significantly lower in patients with well-differentiated HCC than those with moderately differentiated HCC. Recent study demonstrated that IL-8 is an angiogenetic factor. Our present data thus suggest that IL-8 production from HCC may be involved in angiogenesis of this tumor.     

Serum C-reactive protein levels predict survival in hepatocellular carcinoma.

BACKGROUND/AIMS: C-reactive protein (CRP) was recently identified as a prognostic factor for patients with hepatocellular carcinoma (HCC) after surgical resection. We investigated the relationship between the serum levels of high sensitivity CRP (H-CRP) and the prognosis of HCC patients. METHOD: We conducted a cohort study of 90 HCC patients enrolled from 1997 to 1998. All patients were treated and followed for a mean period of 3.2 years. Clinical variables were compared between patients positive for H-CRP (serum H-CRP levels >/=3.0 mg/L, n=47) and those negative for H-CRP (serum H-CRP levels <3.0 mg/L, n=43). We also determined the relationship between serum H-CRP and prognosis in HCC patients. RESULTS: The survival rate of patients of the H-CRP-positive group was lower than that of H-CRP-negative patients. Tumour stage (stages 3 or 4), total bilirubin >/=1.2 mg/dL, albumin (Alb) <3.5 g/dL, des-gamma-carboxy prothrombin >/=40 mAU/mL, positive H-CRP and initial treatment (transcatheter arterial chemoembolization, hepatic arterial infusion chemotherapy or best supportive care) were identified as significant poor prognostic factors by univariate analysis, while positive H-CRP [hazard ratio (HR), 1.58; P=0.048], Alb<3.5 g/dL (HR, 2.10; P=0.004), tumour stage (stages 3 or 4; HR, 3.05; P=0.001) and initial treatment (HR, 1.88; P=0.029) were considered to be significant determinants of poor prognosis by multivariate Cox proportional hazards analysis. CONCLUSIONS: The prognosis of H-CRP-positive patients was poorer compared with H-CRP-negative patients. This study confirmed that H-CRP, like CRP, is a marker of poor prognosis in HCC patients.     

肝癌患者血清microRNA-21水平变化

目的：分析不同肝病患者血清microRNA-21（miR-21）水平,以探讨miR-21对肝细胞癌（HCC）的诊断价值。方法以实时定量逆转录PCR法检测正常人、慢性乙型肝炎（CHB）、肝硬化和HCC患者（各组均为25例）血清miR-21水平,分析miR-21水平与HCC临床病理学特征的关系。结果正常人、CHB和肝硬化患者血清miR-21相对水平分别为（1.1±1.7）、（2.3±2.6）和（2.8±2.5）,而HCC患者为（22.6±4.4）,显著高于前三组（P〈0.001）;HCC患者术后1w和1m血清miR-21相对水平分别为（18.4±3.5）和（3.1±2.7）,均较术前显著降低（P＜0.001）;HCC患者血清miR-21水平与肿瘤大小、癌栓以及HBV感染相关,与肿瘤分化程度、数目和血清AFP水平无相关性。结论 miR-21在HCC患者血清中显著升高,可能作为HCC早期诊断的潜在标志。