Zhi-Hong Xian

SUOX is a promising diagnostic and prognostic biomarker for hepatocellular carcinoma

BACKGROUND & AIMS To investigate diagnostic and prognostic values of sulfite oxidase (SUOX) in patients with hepatocellular carcinoma (HCC) who underwent curative resection. METHODS We investigated immunohistochemically the expression dynamics of SUOX, aldo-ketoreductase family 1 member B10 (AKR1B10) and CD34 at different stages of HCC. The differential diagnostic performance of three markers or their combinations in high-grade dysplastic nodules (HGDNs) and well-differentiated small HCC (WD-sHCC) were investigated by logistic regression models and validated in an independent testing set. Overall survival (OS) and time to recurrence (TTR) were evaluated in 300 patients with HCC as the testing cohort, and validated in 198 patients with HCC. RESULTS SUOX was decreased and AKR1B10 and CD34 were increased with the stepwise progression of hepatocarcinogenesis. For differential diagnosis of WD-sHCC from HGDNs, the sensitivity and specificity of the SUOX+AKR1B10+CD34 combination for WD-sHCC detection were 93.8% and 95.2%, respectively, and overall accuracy was much higher than any of the three individual markers and two marker combinations. In addition, SUOX, but not AKR1B10 and CD34, was an independent prognostic factor for OS and TTR, and showed better correlation with OS and TTR if combined with serum α-fetoprotein (AFP) for both the testing and validation cohorts. CONCLUSIONS SUOX+AKR1B10+CD34 combination could make a substantial contribution to hepatic immunopathological diagnosis to distinguish WD-sHCC from HGDNs. Meanwhile, SUOX combined with serum AFP may predict postoperative outcome and tumor recurrence risk.

Expression of aspartyl beta-hydroxylase and its clinicopathological significance in hepatocellular carcinoma

The human aspartyl beta-hydroxylase is a highly conserved enzyme that hydroxylates epidermal growth factor-like domains in transformation-associated proteins. The aspartyl beta-hydroxylase gene is upregulated in many human malignancies. The purpose of this study was to investigate the expression of aspartyl beta-hydroxylase in hepatocellular carcinoma. Aspartyl beta-hydroxylase mRNA levels were measured in 161 hepatocellular carcinomas and paired nontumoros liver tissues by conventional and real-time RT-PCR. Immunohistochemical staining of aspartyl beta-hydroxylase was performed using EnVision Plus system. The results showed that aspartyl beta-hydroxylase was overexpressed in 150 of 161 hepatocellular carcinomas (93%), including 45 of 48 unifocal small hepatocellular carcinomas (94%). Aspartyl beta-hydroxylase was highly expressed in hepatocellular carcinoma cells in contrast to its low level of expression in non-neoplastic liver cells. The protein expression level of aspartyl beta-hydroxylase in the hepatocellular carcinoma was parallel with the mRNA expression level (r=0.6594, P<0.0001). A significantly higher tumor aspartyl beta-hydroxylase overexpression level was associated with the presence of intrahepatic metastasis and the progression of histological grades. In conclusion, aspartyl beta-hydroxylase is overexpressed frequently in hepatocellular carcinoma, including early-stage small hepatocellular carcinoma, indicating that overexpression of aspartyl beta-hydroxylase plays a role in the development and progression of hepatocellular carcinoma.

Genomic instability in hepatocellular carcinoma revealed by using the random amplified polymorphic DNA method

PurposeTo investigate the genomic instability and their association with clinicopathological characteristics in hepatocellular carcinoma (HCC).MethodsDNA isolated from tumors and corresponding non-cancerous liver tissues of 56 patients with HCC was amplified with ten random 10-mer arbitrary primers by the random amplified polymorphic DNA (RAPD) method.ResultsAll the cases of HCC were demonstrated to have genomic instability by at least one primer. The incidence of genomic instability ranged from 20 to 70% in each case, and 17.9–50% in each primer. Serum AFP concentration, HBV infection, tumor size, histological grade, tumor capsule invasion, as well as intrahepatic metastasis were associated with the genomic instability on certain primers.ConclusionsGenomic instability is a frequent event in HCC. The RAPD is an effective method for the identification and analysis of genomic instability in HCC, and it may provide new information for further evaluating the molecular mechanism of hepatocarcinogenesis.

A novel panel of biomarkers in distinction of small well-differentiated HCC from dysplastic nodules and outcome values

BackgroundDifferential diagnosis of high-grade dysplastic nodules (HGDN) and well-differentiated hepatocellular carcinoma (WDHCC) represents a challenge to experienced hepatic clinicians, radiologists and hepatopathologists.MethodsThe expression profiles of aminoacylase-1 (ACY1), sequestosome-1 (SQSTM1) and glypican-3 (GPC3) in low-grade dysplastic nodules (LGDN), HGDN and WDHCC were assessed by immunohistochemistry. The differential diagnostic performances of these three markers alone and in combination for HGDN and WDHCC were investigated by logistic regression models (HGDN = 21; WDHCC = 32) and validated in an independent test set (HGDN, n = 21; WDHCC n = 24). Postoperative overall survival and time to recurrence were evaluated by univariate and multivariate analyses in an independent set of 500 patients.ResultsACY1, SQSTM1 and GPC3 were differentially expressed in each group. For the differential diagnosis of WDHCC from HGDN, the sensitivity and specificity of the combination of ACY1 + SQSTM1 + GPC3 for detecting WDHCC were 93.8% and 95.2% respectively in the training set, which were higher than any of the three two-marker combinations. The validities of the four diagnostic models were further confirmed in an independent test set, and corresponding good sensitivity and specificity were observed. Interestingly, GPC3 expression in HCC tissues combined with serum α-fetoprotein (AFP) was found to be an independent predictor for overall survival and time to recurrence.ConclusionsACY1 + SQSTM1 + GPC3 combination represents a potentially valuable biomarker for distinguishing between WDHCC and HGDN using immunohistochemistry. Meanwhile, low GPC3 staining combined with positive serum AFP may play a practical role in predicting poor postoperative outcome and high tumor recurrence risk.

Using loss of heterozygosity of microsatellites to distinguish high-grade dysplastic nodule from early minute hepatocellular carcinoma

BACKGROUND It is practical significant to seek new applicable adjuvant diagnostic biomarkers to differentiate high-grade dysplastic nodule (HGDN) from well-differentiated minute hepatocellular carcinoma (w-MHCC) due to their closely overlapping morphology. METHODS In the present study, by using microdissection-based paraffin-embedded tissues, loss of heterozygosity (LOH) patterns of a panel of 22 microsatellite (MS) markers was examined in 8 HGDN, 14 w-MHCC (≤1 cm) and 35 larger HCC (LHCC, >1 cm). RESULTS The results revealed a stepwise increasing fractional allelic loss from HGDN, w-MHCC and LHCC (0.166±0.141, 0.377±0.198, 0.471±0.264, respectively, P=0.005). Loci-specific analyses showed that LOH on D4S415 (66.7% vs 0.0%, P=0.04), D1S507 (50.0% vs 0.0%, P=0.098), and D9S1752 (50.0% vs 0.0%, P=0.33) occurred more frequently than 50% in w-MHCC, but not in HGDN. On the other hand, LOH on D17S960, D17S1796 and D9S1749 occurred in HGDN, but not in w-MHCC. When compared with w-MHCC, LHCC had a higher LOH frequency on D17S720 (73.9% vs 36.4%, P=0.04), D17S960 (68.8% vs 0.0%, P=0.03) and D17S1796 (81.8% vs 0.0%, P=0.01). CONCLUSIONS The present study suggests MS-LOH is a simple and specific assay for routinely diagnostic pathology. We recommend that D4S415, D1S507, D9S1752, D17S960, D17S1796 and D9S1749 can be used as the first-line markers for differential diagnosis between HGDN and w-MHCC, and D9S1748, D17S921 and D17S520 with a LOH frequency of 40%-50% in w-MHCC, but netative in HGDN, can be regarded as the second-line candidate markers.

Expression and localization of aquaporin-1 in human cirrhotic liver

We investigated the expression and localization of aquaporin-1 (AQP-1) in hepatitis B virus (HBV)-associated cirrhotic human liver tissues. The expression of AQP-1 at the protein and mRNA levels was analyzed by immunohistochemistry, quantitative real-time polymerase chain reaction (qPCR) and Western blotting in normal and HBV-associated cirrhotic human liver tissues. The correlation with the expression of CK19, CK7 and AQP-1 was also compared. AQP-1 staining was strongly and uniformly positive in mature bile ducts, isolated hepatic progenitor cells (HPCs) and ductular reactions. Scattered intermediate hepatocyte-like cells expressed AQP-1, which are often intimately associated with CK7 positive hepatocytes. However, the number of AQP-1+ intermediate hepatocyte-like cells was lower than that of CK7+ cells, and such positivity was rarely seen on stains for CK19. When compared with normal liver tissues, AQP-1 was overexpressed at both the mRNA and protein levels in the cirrhotic liver tissues. AQP-1 was overexpressed in the cirrhotic liver tissues. AQP-1, similar to CK19, might be a more specific and more sensitive marker than CK7 for the identification of HPCs.

Angiogenesis and lymphangiogenesis in sporadic hepatic angiomyolipoma.

Angiogenesis and lymphangiogenesis are critical processes for tumor growth, invasion, and metastasis. The present study aimed to investigate the distribution and clinical significance of angiogenesis and lymphangiogenesis in hepatic angiomyolipoma (AML). We performed immunohistochemical staining for endothelial cell markers (CD34 and podoplanin) on 80 cases of sporadic hepatic AMLs. Microvessel density (MVD) and lymphatic vessel density (LVD) were determined in intratumoral and peritumoral regions and adjacent non-tumorous liver tissues. All hepatic AMLs showed positive staining for CD34 and podoplanin. Intratumoral and peritumoral MVDs and LVDs were significantly higher than those in adjacent liver tissues (P<0.001). No statistical difference in both MVD and LVD was found between intratumoral and peritumoral areas. Large tumors (>5cm) had a significantly increased MVD and LVD as compared with smaller tumors. A significant positive correlation was found between average LVDs and MVDs (r=0.567, P<0.001), and LVDs were a relatively lower event as compared with MVDs. Double immunostaining revealed that no neoplastic cells positive for HMB-45, an antibody reacting with melanosome-associated antigen, were concurrently immunoreactive for endothelial cell markers. In conclusion, intratumoral and peritumoral angiogenesis and lymphangiogenesis commonly occur in hepatic AMLs, thus representing potential therapeutic targets for this disease.