Wei-Dong Jia

Ligation of TLR2 by Versican: A Link Between Inflammation and Metastasis

Versican, a large extracellular matrix proteoglycan, accumulates both in tumor stroma and cancer cells. It participates in cell adhesion, migration, and angiogenesis, all features of invasion and metastasis. However, the mechanism(s) whereby versican promotes cancer invasion and metastasis is not yet fully understood. A recent study has documented that versican can activate tumor-infiltrating myeloid cells through toll-like receptor (TLR) 2 and its co-receptors TLR6 and CD14 and elicit the production of proinflammatory cytokines including TNF-alpha that enhance tumor metastasis. As both resident fibroblasts and endothelial cells (ECs) also express functional TLR2 and its co-receptors, we hypothesized that, in addition to myeloid cells, versican may trigger the activation of both fibroblasts and ECs. Of interest, TLR2-mediated activation of EC and fibroblast has been observed to increase the secretion of interleukin-8, a proinflammatory CXC chemokine that potentiates neutrophil infiltration and angiogenesis, as well as metastatic growth. Ligation of TLR2 by versican appears to be directly involved in the activation of multiple types of cells in tumor stroma and the induction of inflammatory cytokine secretion, providing a link between inflammation and cancer metastasis. Accordingly, antagonists of versican and TLR2 restrain the activation of tumor stromal cells, which may offer a novel approach to cancer therapy by targeting tumor microenvironment.

Effects of interferon treatment on development and progression of hepatocellular carcinoma in patients with chronic virus infection: A meta‐analysis of randomized controlled trials

Available literature on the effects of interferon (IFN) treatment on development and progression of hepatocellular carcinoma (HCC) in patients with chronic virus infection reports controversial results. The primary objective of this meta‐analysis was to evaluate the effect of IFN on HCC risk in patients with chronic hepatitis C virus (HCV) or hepatitis B virus (HBV) infection; IFNs efficacy on local tumor progression and survival of advanced HCC patients was also assessed. All randomized controlled trials (RCTs) comparing IFN with no antiviral treatment were selected. Finally, we identified 11 RCTs including 1,772 patients, who met our inclusion criteria to perform this meta‐analysis. Our analysis results showed that IFN significantly decreased the overall HCC incidence in HCV‐infected patients [relative risk (RR) = 0.39; 95% confidence interval (CI) = 0.26–0.59; p = 0.000], subgroup analysis indicated that IFN decreased HCC incidence in HCV‐related cirrhotic patients evidently (RR = 0.44; 95% CI = 0.28–0.68; p = 0.000); but HCC incidence in nonresponders to initial antiviral therapy did not reduce by maintenance IFN therapy (RR = 0.96; 95% CI = 0.59–1.56; p = 0.864). Analysis results also demonstrated that IFN did not significantly affect the overall rate of HCC in HBV‐infected patients although there was a trend favoring IFN therapy (RR = 0.23; 95% CI = 0.05–1.04; p = 0.056). Besides, IFN did not improve one‐year overall survival of advanced HCC patients significantly (RR = 1.61; 95% CI = 0.96–2.69; p = 0.072); however, a quantitative analysis on local tumor progression could not be performed owing to lack of unified definitions among trials included in our study. By this meta‐analysis, we conclude that IFN therapy is effective in reducing overall HCC risk in chronic HCV‐infected patients; using it in this subpopulation seems promising, but its administration in other subpopulations still requires further exploration.

Prognostic significance of osteopontin in hepatocellular carcinoma: A meta‐analysis

Osteopontin (OPN) has been implicated in tumor development and progression for several years. However, the prognostic value of OPN overexpression in patients with hepatocellular carcinoma (HCC) remains controversial. We performed a meta‐analysis to assess the relationship between OPN overexpression and clinical outcome of HCC. A meta‐analysis of seven studies (1,158 patients) was carried out to evaluate the association between OPN and overall survival (OS) and disease‐free survival (DFS) in HCC patients. The correlation between OPN and tumor vascular invasion or other invasion‐related parameters was also assessed. Data were synthesized with random effect model of DerSimonian and Laird, hazard ratio (HR) or odds ratio (OR) with its 95% confidence interval (CI) was used as the effect size estimate. Our analysis results indicated that high OPN expression predicted poor OS (HR: 1.37, 95% CI: 1.21–1.55) and DFS (HR: 1.62, 95% CI: 1.24‐2.11) of HCC. OPN overexpression tended to be associated with the presence of tumor vascular invasion (OR: 1.93, 95% CI: 0.97–3.84) and advanced tumor grade (OR: 1.74, 95% CI: 0.95–3.18). By this study, we conclude that OPN overexpression indicates a poor prognosis for patients with HCC, it may also have predictive potential for HCC invasion and metastasis.

Effects of interferon alpha treatment on recurrence and survival after complete resection or ablation of hepatocellular carcinoma: a meta-analysis of randomized controlled trials.

Available literature on the benefit of interferon alpha (IFN‐α) as adjuvant postsurgical or ablative treatment of hepatocellular carcinoma reports discordant results. By meta‐analysis of the available data, we evaluated the effects of IFN‐α on recurrence and survival after complete resection or ablation of hepatocellular carcinoma. All randomized controlled trials comparing IFN‐α with placebo or no treatment after tumor resection or ablation were selected. Finally, 6 studies published in 2001 or later with a total of 600 patients were included in this meta‐analysis. Data on postsurgical or ablative early recurrence and 1 year survival of hepatocellular carcinoma in IFN‐α treated and untreated patients were extracted from each study. Proportions were combined, and the odds ratio (OR) with its 95% confidence interval (CI) was used as the effect size estimate. Analysis results show that IFN‐α significantly decreased postsurgical or ablative overall early recurrence (OR = 0.62; 95% CI = 0.42–0.93; p = 0.02) and improved overall 1 year survival (OR = 3.14; 95% CI = 1.79–5.52; p < 0.0001). Subgroup analyses show that IFN‐α decreased postsurgical early recurrence (OR = 0.58; 95% CI = 0.37–0.91; p = 0.02) and improved 1 year survival (OR = 3.19; 95% CI = 1.80–5.67; p < 0.0001) evidently. Subgroup analyses also show that IFN‐α reduced early recurrence after resection without pre‐resection ablation therapy (OR = 0.58; 95% CI = 0.37–0.91; p = 0.02) and improved 1 year survival (OR = 3.83; 95% CI = 2.01–7.27; p < 0.0001). These results suggest that IFN‐α treatment could significantly decrease early recurrence and improve 1 year survival of patients with hepatocellular carcinoma after complete resection or ablation. The use of IFN‐α as adjuvant postsurgical or ablative treatment seems promising but requires further study.

Expression and Correlation of Hypoxia-inducible Factor-1α, Vascular Endothelial Growth Factor and Microvessel Density in Experimental Rat Hepatocarcinogenesis

An experimental rat hepatocellular carcinoma (HCC) model was established using diethylnitrosamine and N-nitrosomorpholine to induce carcinogenesis in Sprague-Dawley rats. During hepatocarcinogenesis, seven rats were sacrificed at 0, 4, 8, 12 and 16 weeks and 10 rats were sacrificed at 20 weeks. The levels of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) protein and mRNA were examined by immunohistochemistry, Western blot and semi-quantitative reverse transcriptase-polymerase chain reaction at different stages in the rat HCC model. Twenty weeks after induction of hepatocarcinogenesis, the expression of HIF-1α and VEGF protein and mRNA significantly increased compared with week 0. Microvessel density (MVD) increased considerably once liver cancer developed. There was a significant positive correlation between MVD and both HIF-1α and VEGF, and between HIF-1α and VEGF levels. These results suggest that HIF-1α and VEGF play important roles in tumour occurrence and development during rat hepatocarcinogenesis, possibly through promoting tumour angiogenesis.

Combined Inhibitory Effects of Celecoxib and Fluvastatin on the Growth of Human Hepatocellular Carcinoma Xenografts in Nude Mice

This study was designed to investigate the in vivo growth inhibitory effects of celecoxib, a cyclo-oxygenase-2 inhibitor, and fluvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, on the hepatocellular carcinoma (HCC) cell line, BEL-7402. Athymic nude mice implanted with BEL-7402 cells were given celecoxib and fluvastatin, either alone or in combination, and the effect of treatment on tumour growth was evaluated after 6 weeks. The combination of celecoxib and fluvastatin enhanced inhibition of tumour growth, induction of apoptosis, inhibition of tumour cell proliferation, and inhibition of tumour angiogenesis compared with either treatment alone. The combination of celecoxib and fluvastatin also increased levels of the cyclin-dependent kinase inhibitor p21Waf1/Cip1, decreased levels of p-Akt, myeloid cell leukaemia-1 (Mcl-1) and survivin protein, but had no effect on Akt protein levels in tumours. These results suggest that celecoxib combined with fluvastatin would be more efficacious for the treatment of HCC than either treatment alone and this combination of therapy warrants further research.

Overexpression of Mortalin in hepatocellular carcinoma and its relationship with angiogenesis and epithelial to mesenchymal transition

Mortalin is highly expressed in a variety of human tumors and associated with tumor metastasis. However, the relationship among the overexpression of Mortalin, epithelial to mesenchymal transition (EMT) and neovascularization is largely unknown. The aim of the present study was to investigate the expression of Mortalin in human HCC cell lines, clinical HCC specimens and its association with angiogenesis and EMT. The results of our study showed that the expression levels of Mortalin in cell lines with higher metastatic potential were significantly higher compared to those with lower metastatic potential. Compared with paracarcinomatous tissues and normal liver tissues, the expression of Mortalin was significantly increased in HCC tumor tissues. The expression of Mortalin was correlated with invasion and metastasis, Edmondson grade and TNM stage. A significant positive correlation was found between the expression of Mortalin and Vimentin, and tumors with high expression of Mortalin had a tendency to higher MVD compared to those with low expression of Mortalin. Using shRNA-mediated Mortalin knockdown, we found that decreased expression of Mortalin was accompanied by a reduction of Vimentin expression. Our findings demonstrated that the overexpression of Mortalin is correlated with the metastatic phenotype of HCC cells and can promote EMT, but cannot induce angiogenesis in HCC. The decreased expression of Mortalin is accompanied by an inhibition of EMT in the HCC cell lines.

Prognostic value of polarized macrophages in patients with hepatocellular carcinoma after curative resection.

As the most predominant tumour‐infiltrating immune cells, tumour‐associated macrophages (TAMs) are significant for fostering tumour growth, progression and metastasis. CD68‐positive TAMs display dissimilarly polarized programmes comprising CD11c‐positive pro‐inflammatory macrophages (M1) and CD206‐positive immunosuppressive macrophages (M2). The aim of this study is to determine the prognostic significance of diametrically polarized TAMs in hepatocellular carcinoma (HCC) and their application to risk stratification of patients according to their specific prognostic values. This study included 80 consecutive patients with HCC, and we evaluated diametrically polarized functional status of macrophages by immunohistochemical staining of CD68, CD11c and CD206. Prognostic values and clinicopathologic features were assessed in these patients. High versus low CD11c‐positive TAM density (P = 0.005) and low versus high CD206‐positive TAM density (P = 0.002) were associated with better overall survival, whereas CD68‐positive TAM density had no prognostic significance (low versus high, P = 0.065). Furthermore, the presence of these positive staining macrophages did not show any prognostic significance for recurrence‐free survival (all P > 0.05). Multivariate Cox regression analysis identified CD11c‐positive and CD206‐positive TAMs as an independent prognostic factor (P < 0.001, P = 0.031, respectively). Intratumoural infiltration of diametrically polarized TAMs, a novel identified independent prognostic factor for survival in patients with HCC, could be combined with the TNM stage and the Barcelona Clinic Liver Cancer stage to improve a risk stratification system.

Kindlin-2: A novel prognostic biomarker for patients with hepatocellular carcinoma

The association of aberrant expression of Kindlin-2 with tumor progression has been reported in recent years. The purpose of this study was to investigate the expression of Kindlin-2 in hepatocellular carcinoma (HCC), and to evaluate its clinical and prognostic significance. The mRNA and protein levels of Kindlin-2 in HCC and adjacent non-cancerous tissues were examined by real-time PCR and western blotting. The relationships between Kindlin-2 expression, clinicopathological features and postoperative survival of HCC patients were also evaluated. Kindlin-2 expression was higher in HCC tissues as compared to adjacent non-cancerous tissues at both mRNA and protein levels (P<0.05, respectively). Positive expression of Kindlin-2 was significantly correlated with larger tumor size (P=0.034), capsular invasion (P=0.009), microvascular invasion (P=0.028) and poor prognosis of HCC patients (P<0.001). Moreover, multivariate survival analysis identified Kindlin-2 as an independent prognostic factor for overall and disease-free survival of HCC patients (P=0.018 and 0.001, respectively). Taken together, our findings suggested that Kindlin-2 was highly expressed in HCC tissues and was closely related to clinical progression. Therefore, Kindlin-2 protein could be a potential biomarker for predicting poor prognosis of HCC patients after surgery.

Embryonic Morphogen Nodal Is Associated with Progression and Poor Prognosis of Hepatocellular Carcinoma

Background Nodal, a TGF-β-related embryonic morphogen, is involved in multiple biologic processes. However, the expression of Nodal in hepatocellular carcinoma (HCC) and its correlation with tumor angiogenesis, epithelial-mesenchymal transition, and prognosis is unclear. Methods We used real-time PCR and Western blotting to investigate Nodal expression in 6 HCC cell lines and 1 normal liver cell line, 16 pairs of tumor and corresponding paracarcinomatous tissues from HCC patients. Immunohistochemistry was performed to examine Nodal expression in HCC and corresponding paracarcinomatous tissues from 96 patients. CD34 and Vimentin were only examined in HCC tissues of patients mentioned above. Nodal gene was silenced by shRNA in MHCC97H and HCCLM3 cell lines, and cell migration and invasion were detected. Statistical analyses were applied to evaluate the prognostic value and associations of Nodal expression with clinical parameters. Results Nodal expression was detected in HCC cell lines with high metastatic potential alone. Nodal expression is up-regulated in HCC tissues compared with paracarcinomatous and normal liver tissues. Nodal protein was expressed in 70 of the 96 (72.9%) HCC tumors, and was associated with vascular invasion (P = 0.000), status of metastasis (P = 0.004), AFP (P = 0.049), ICGR15 (indocyanine green retention rate at 15 min) (P = 0.010) and tumor size (P = 0.000). High Nodal expression was positively correlated with high MVD (microvessal density) (P = 0.006), but not with Vimentin expression (P = 0.053). Significantly fewer migrated and invaded cells were seen in shRNA group compared with blank group and negative control group (P<0.05). High Nodal expression was found to be an independent factor for predicting overall survival of HCC. Conclusions Our study demonstrated that Nodal expression is associated with aggressive characteristics of HCC. Its aberrant expression may be a predictive factor of unfavorable prognosis for HCC after surgery.