Hepatocellular carcinoma screening and diagnosis.

All the major liver disease societies have recommended screening for hepatocellular carcinoma (HCC). The target population for HCC screening has been defined by cost-efficacy analyses and by risk scores. Risk scores have been developed for patients with hepatitis B, regardless of the presence of cirrhosis, and for other patients with cirrhosis. Screening is with ultrasound; however, in Asia biomarkers are also used. The additional value of biomarkers has not been demonstrated. The ideal screening interval is 6 months; in Japan shorter intervals are used. Screening detects small lesions that require confirmation of HCC. There are radiological criteria that help determine whether a biopsy is necessary. Special stains can determine whether a lesion that closely resembles normal or dysplastic tissue is HCC. All these tools should be used in the management of patients undergoing HCC screening.

The transcription factor SALL4 regulates stemness of EpCAM-positive hepatocellular carcinoma.

BACKGROUND & AIMS: Recent evidence suggests that hepatocellular carcinoma can be classified into certain molecular subtypes with distinct prognoses based on the stem/maturational status of the tumor. We investigated the transcription program deregulated in hepatocellular carcinomas with stem cell features. METHODS: Gene and protein expression profiles were obtained from 238 (analyzed by microarray), 144 (analyzed by immunohistochemistry), and 61 (analyzed by qRT-PCR) hepatocellular carcinoma cases. Activation/suppression of an identified transcription factor was used to evaluate its role in cell lines. The relationship of the transcription factor and prognosis was statistically examined. RESULTS: The transcription factor SALL4, known to regulate stemness in embryonic and hematopoietic stem cells, was found to be activated in a hepatocellular carcinoma subtype with stem cell features. SALL4-positive hepatocellular carcinoma patients were associated with high values of serum alpha fetoprotein, high frequency of hepatitis B virus infection, and poor prognosis after surgery compared with SALL4-negative patients. Activation of SALL4 enhanced spheroid formation and invasion capacities, key characteristics of cancer stem cells, and up-regulated the hepatic stem cell markers KRT19, EPCAM, and CD44 in cell lines. Knockdown of SALL4 resulted in the down-regulation of these stem cell markers, together with attenuation of the invasion capacity. The SALL4 expression status was associated with histone deacetylase activity in cell lines, and the histone deacetylase inhibitor successfully suppressed proliferation of SALL4-positive hepatocellular carcinoma cells. CONCLUSIONS: SALL4 is a valuable biomarker and therapeutic target for the diagnosis and treatment of hepatocellular carcinoma with stem cell features.

Concanavalin A-immobilized magnetic nanoparticles for selective enrichment of glycoproteins and application to glycoproteomics in hepatocelluar carcinoma cell line.

Protein glycosylation is one of the most important PTMs in biological organism. Lectins such as concanavalin A (Con A) have been widely applied to N-glycosylated protein investigation. In this study, we developed Con A-immobilized magnetic nanoparticles for selective separation of glycoproteins. At first, a facile immobilization of Con A on aminophenylboronic acid-functionalized magnetic nanoparticles was performed by forming boronic acid-sugar-Con A bond in sandwich structure using methyl alpha-D-mannopyranoside as an intermedium. The selective capture ability of Con A-modified magnetic nanoparticles for glycoproteins was tested using standard glycoproteins and cell lysate of human hepatocelluar carcinoma cell line 7703. In total 184 glycosylated sites were detected within 172 different glycopeptides corresponding to 101 glycoproteins. Also, the regeneration of the protein-immobilized nanoparticles can easily be performed taking advantage of the reversible binding mechanism between boronic acid and sugar chain. The experiment results demonstrated that Con A-modified magnetic nanoparticles by the facile and low-cost synthesis provided a convenient and efficient enrichment approach for glycoproteins, and are promising candidates for large-scale glycoproteomic research in complicated biological samples.

Small hepatocellular carcinoma: assessment with T1-weighted spin-echo magnetic resonance imaging with and without fat suppression.

OBJECTIVE: We examined the detectability of small hepatocellular carcinomas (HCCs) and the factors that affect hyperintensity of small HCC on T1-weighted images (T1W) by using T1-weighted fat-suppressed images (T1FS). METHODS: Thirty-nine HCCs (29 patients) measuring 30 mm or less were enrolled. The mean size of HCCs was 21.0+/-4.9 mm. Spin-echo T1W, T2-weighted images (T2W), and T1FS were obtained using a 1.5 T system. We evaluated the detectability in each sequence by receiver-operating-characteristic (ROC) analysis and the tumor-to-hepatic parenchyma contrast-to-noise ratio (CNR), the variance in the detectability among all interpreters with each sequence, and the presence or absence of improvement in the detectability by interpreting T1FS in addition to conventional T1W plus T2W. The contents of fat, copper, and iron in histologically diagnosed HCCs showing hyperintensity on both T1W and T1FS were measured. For determination of heavy metals, we used a particle induced X-ray emission analytical instrument. RESULTS: ROC analyses revealed that T1FS were superior to T1W and T2W in detecting small HCCs (0.900+/-0.017 for T1FS, 0.859+/-0.019 for T1W, and 0.745+/-0.030 for T2W). The detectability by interpreting T1FS in addition to conventional T1W plus T2W was improved (0.931+/-0.013 for the conventional images and 0.973+/-0.008 for the conventional images plus T1FS, P<0.001). The detected lesions on T1FS demonstrated favorable CNR values. The copper content in the cancer and the ratio of the copper content in the cancer to that in the non-cancerous tissue were 275.4+/-219.0 microg/g dry weight, 6.9+/-5.5 in HCCs showing hyperintensity on both T1W and T1FS. Both were significantly higher (P<0.05). CONCLUSION: T1FS showed excellent sensitivity and specificity in detecting small HCCS irrespective of the experience of interpreters. The use of T1FS suggested the involvement of copper might be one of the factors in hyperintensity of HCCs on T1W.

The assessment of proliferating cell nuclear antigen immunohistochemical staining in small hepatocellular carcinoma and its relationship to histologic characteristics and prognosis.

BACKGROUND: A precise prognostic factor for small hepatocellular carcinoma (HCC), the diagnosis of which recently has increased in incidence because of the development of diagnostic imaging techniques, is desirable. It has been reported that proliferating cell nuclear antigen (PCNA) would be related to proliferating cells, and thus the PCNA labeling index may provide useful information about the biologic behavior of small HCC. METHODS: An assessment was made of proliferative activity by immunohistochemical staining using a monoclonal antibody against PCNA in 46 nodules of HCC less than 3 cm in diameter resected from 44 patients. A correlation between PCNA labeling index and clinicopathologic findings or prognosis was sought. RESULTS: The mean labeling index was 18.7% in HCC and 1.9% in nontumor liver tissue. The labeling index corresponded to the degree of histologic differentiation, and the labeling index of well differentiated HCC was significantly lower (P < 0.05) than that of moderately or poorly differentiated HCC. The incidence of capsule formation in the high labeling index group (labeling index > or = 20%) was significantly higher (P < 0.05) than that in the low labeling index group (labeling index < 20%). A high incidence of capsular and vascular invasion was found in the high labeling index group. The survival rate after resection was significantly higher (P < 0.05) and the recurrence rate significantly lower (P < 0.05) in the low labeling index group than in the high labeling index group. CONCLUSIONS: The PCNA labeling index was shown to be closely related to histologic characteristics, and proved to be a useful indicator of recurrence and survival in small HCC.