The expression of CXCR4 and its relationship with matrix metalloproteinase‐9/vascular endothelial growth factor in esophageal squamous cell cancer

Esophageal cancer (EC) is a highly aggressive neoplasm with poor prognosis. The main reason for this disappointing outcome is the strong behavior of esophageal cancer cell's invasion and metastasis. CXC chemokine receptor 4 (CXCR4) was found to be expressed in many tumors and significantly correlated with invasion, angiogenesis, metastasis, and prognosis. In the present study, we investigated the expressions of CXCR4, matrix metalloproteinase‐9 (MMP‐9), and vascular endothelial growth factor (VEGF) in esophageal squamous cell cancer (ESCC) and analyzed the relationship among the three proteins. Sections of paraffin‐embedded tissues were obtained from 127 patients with ESCC undergoing esophagectomy at Zhongshan Hospital, Fudan University in 2005. The CXCR4, MMP‐9, and VEGF expressions in EC tissues were evaluated according to the immunohistochemical staining area and intensity. The correlations between patients' prognosis and covariates were analyzed by Kaplan–Meier method (univariate analysis) and Cox regression (multivariate analysis). The overall expression rate of CXCR4, MMP‐9, and VEGF was 88.2%, 93.7%, and 79.5%, respectively. CXCR4 expression was significantly associated with tumor grade, tumor size, tumor depth, regional lymph node metastasis, and tumor, node, metastasis (TNM) stage (P < 0.05). MMP‐9 expression was significantly associated with age and tumor grade (P < 0.05). VEGF expression was significantly associated with tumor grade, tumor depth, and TNM stage (P < 0.05). CXCR4 expression was positively correlated with MMP‐9 expression (P < 0.01, r= 0.365) and VEGF expression (P < 0.01, r= 0.380). However, there was no significant correlation between MMP‐9 and VEGF expression (P > 0.05). In univariate analysis, CXCR4 expression, tumor size, tumor depth, lymph node metastasis, and TNM stage were correlated with patients' prognosis (P < 0.05); in multivariate analysis, tumor size and lymph node metastasis were the independent factors of poor prognosis. CXCR4 was highly expressed in ESCC and correlated with MMP‐9, VEGF, clinicopathological features and prognosis. We speculated CXCR4 play an important role during the progression of this disease and there might be some regulatory mechanism existing between CXCR4 and MMP‐9/VEGF in ESCC.     

Overexpression of MMP‐2 and MMP‐9 in esophageal squamous cell carcinoma

Matrix metalloproteinases (MMPs) are known to play important roles in extracellular matrix remodeling during the process of tumor invasion and metastasis. However, little is known about their role in esophageal squamous cell carcinoma (ESCC). Expression of MMP‐2 and MMP‐9 in ESCC was detected in our research. Tissue microarray chip was prepared, consisting of 58 cases of ESCC and corresponding esophageal epithelium tissues. MMP‐2 and MMP‐9 were examined by immunohistochemistry. Overexpression of MMP‐2 and MMP‐9 was found in ESCC (42.1 and 60.3%, respectively), compared with paired distal normal esophageal tissues (22.9 and 8.9%, respectively). Expression of MMP‐2 in ESCC was significantly associated with the tumor invasion depth, tumor‐node‐metastasis stages, and lymph node metastasis. MMP‐2 and MMP‐9 may play important roles in carcinogenesis, and MMP‐2 may act as a biological marker of invasion and lymph node metastasis in ESCC.     

Identification of two portal vein tumor thrombosis associated proteins in hepatocellular carcinoma: protein disulfide-isomerase A6 and apolipoprotein A-I

Background and Aim: Portal vein tumor thrombus (PVTT) is one of the factors that can affect prognosis and survival of hepatocellular carcinoma (HCC). In the present study, we aimed to find out some biomarkers associated with vascular invasion features of HCC with the method of comparative proteomic analysis. Methods: The proteins were extracted from a pair of HCC tissues with PVTT and without PVTT, and then separated by two‐dimensional polyacrylamide gel electrophoresis. Differentially expressed protein spots were identified by matrix assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS). Further analysis of two proteins were completed using real‐time fluorescence quantitative polymerase chain reaction and western‐blot in 40 HCC tissues with PVTT (n = 20) and without PVTT (n = 20). Results: Among 465 protein spots displayed on the gels, 33 unique proteins (> twofold change, P < 0.01) were identified, including 24 upregulated in HCC tissue without PVTT and nine upregulated in HCC tissue with PVTT. The real‐time fluorescence quantitative PCR showed no statistically significant difference between HCC tissues with PVTT and without PVTT for mRNA expressions of protein disulfide‐isomerase, A6 (PDI A6) (P = 0.137) and apolipoprotein A‐I (Apo A‐I) (P = 0.718). However, compared with HCC tissues without PVTT, protein expression of PDI A6 was higher in HCC tissues with PVTT (P < 0.001), while protein expression of Apo A‐I was lower in HCC tissues with PVTT (P = 0.012). Conclusions: PDI A6 and Apo A‐I are closely related to vascular invasion feature of HCC.     

Reduced N-cadherin expression is associated with metastatic potential and poor surgical outcomes of hepatocellular carcinoma

Background and Aim: N‐cadherin (N‐cad), one of the classic cadherins, has been reported to be involved in tumor metastasis in some types of tumors. This study aims to investigate the expression status of N‐cad in hepatocellular carcinoma (HCC) and the correlation between N‐cad expression and metastatic potential, as well as the surgical outcomes of HCC. Methods: N‐cad expression in HCC and adjacent liver tissues, as well as normal liver tissues, was studied by immunohistochemistry and Western blot, and the relationship between N‐cad expression and the clinicopathological features of HCC was evaluated. By using RNA interference technique, the correlation of N‐cad expression and metastatic potential was investigated by downregulating N‐cad expression in HCCLM3 cells, and the effects of N‐cad downregulation on cell aggregation, migration, and invasion were then analyzed. Furthermore, the correlation between N‐cad expression and the surgical outcomes of a cohort of HCC patients was analyzed. Results: In liver tissues, N‐cad was strongly expressed on cell–cell boundaries, whereas various reduced‐expression patterns were observed in tumors. Of 64 HCC, 34 (53%) tumors showed reduced N‐cad expression, compared with their adjacent liver tissues. The decreased expression of N‐cad was significantly correlated with poorer tumor differentiation (P = 0.001) and vascular invasion (P = 0.003). N‐cad knockdown in HCCLM3 cells resulted in decreased cell aggregation and increased cell migration and invasion. The decreased expression of N‐cad in HCC was significantly associated with shorter postoperative disease‐free survival (P = 0.039). Conclusions: N‐cad expression is decreased in HCC, and the downregulation of N‐cad is associated with the metastatic potential of HCC and poorer surgical prognosis.     

Aldehyde dehydrogenase 1 is associated with recurrence‐free survival but not stem cell‐like properties in hepatocellular carcinoma

Aim: It has been reported that aldehyde dehydrogenase 1 A1 (ALDH1) could be not only a normal stem cell marker but also a cancer stem cell marker. ALDH1 expression could be a predictor of poor prognosis in a wide range of cancers. However, the role of ALDH1 in hepatocellular carcinoma (HCC) remains unclear. Method: We conducted loss‐of‐function assays for ALDH1 by using short‐hairpin RNA in HCC cells and evaluated the correlation between ALDH1 expression and clinicopathological features based on immunohistochemical assessment of 49 primary HCC tissues. Results: Neither cell proliferation nor the anchorage‐independent sphere formation ability of HCC cells were altered after ALDH1 knockdown. Flow cytometric analyses revealed that ALDH1 knockdown showed no remarkable change in the proportion of epithelial cell adhesion molecule (EpCAM)⁺ tumor‐initiating cells. Although non‐tumor tissues in primary HCC samples diffusely and homogenously expressed ALDH1 at low levels, tumor tissues contained cells with high levels of ALDH1 expression at varying frequencies. Primary HCC samples were categorized as ALDH1‐high or ALDH1‐low based on the percentage of ALDH1‐overexpressing cells. ALDH1‐high HCC was characterized by low serum levels of α‐fetoprotein (P < 0.01) and well‐differentiated pathology (P = 0.03). Multivariate analysis showed that high ALDH1 expression was a favorable prognostic factor in recurrence‐free survival of HCC (P = 0.02). Conclusion: Our findings show that ALDH1 expression has little association with stem cell‐like features in HCC cells. ALDH1 might function as a differentiation marker rather than a stem cell marker in HCC.     

Overexpression of Yes‐associated protein and its association with clinicopathological features of hepatocellular carcinoma: A meta‐analysis

Background

Yes‐associated protein (YAP) overexpression is reported to be associated with risk of hepatocellular carcinoma (HCC) but current studies have not explored the relationship between YAP expression with HCC clinicopathological features.

Methods

To assess these associations, a meta‐analysis was performed which included four eligible studies including 391 HCC cases and 334 controls. There were eight eligible studies to investigate the association between YAP expression in HCC and clinicopathological features of liver cancer patients. Literature was obtained from PubMed, Embase, Wangfang and China National Knowledge Infrastructure.

Results

Analysis indicated that YAP expression in HCC was greater than in adjacent non‐tumour tissue (odds ratio [OR], 15.80, 95% confidence interval [CI], 10.53‐23.70, P<.00001; heterogeneity=.30). YAP overexpression in HCC was significantly associated with vascular invasion (OR, 2.21, 95% CI, 11.64‐2.97, P<.00001, heterogeneity=.10), less cellular differentiation (OR, 2.38, 95% CI, 1.61‐3.51, P<.00001, heterogeneity=.333), tumours larger than 5 cm (OR, 2.52, 95% CI, 1.75‐3.62, P<.00001; heterogeneity=.17) and TNM tumour stage I + II (OR, 0.44, 95% CI, 0.28‐0.75, P=.00003, heterogeneity=.12).

Conclusions

Overexpression of YAP contributes to HCC formation, and its overexpression is associated with vascular invasion, low cellular differentiation tumours larger than 5 cm and TNM tumour stage III + IV.     

KAI1 gene suppresses invasion and metastasis of hepatocellular carcinoma MHCC97‐H cells in vitro and in animal models

Background: Downregulation of KAI1 gene expression has been found in many types of cancer cells and is closely related to cancer invasion and metastasis. This study was aimed at investigating the effects and possible underlying mechanisms of KAI1 gene on invasion and metastasis of human hepatocellular carcinoma (HCC). Methods: The invasive ability, visco‐elastic properties and cell adhesion forces were analysed in different HCC cells originating from the MHCC97‐H cell line transfected with either the sense or the antisense KAI1 expression plasmid. Tumuorigenicity, metastatic abilities, extracellular matrix (ECM) and intercellular adhesion molecule‐1 (ICAM‐1) expression were also evaluated in the nude mouse models of the xenografted and orthotopic liver cancer cells. Results: Compared with their parental cells, in the HCC cells transfected with the sense KAI1 gene, the invasive ability in vitro was significantly decreased (P<0.01); the cellular elastic coefficients K₁, K₂ and μ were significantly higher (P<0.05); the cells adhesion forces to fibronectin were significantly lower (P<0.01). The sense KAI1 gene transfection into the cancer cells also inhibited their invasion and lung metastasis in the orthotopic liver cancer nude mice. However, the opposite changes were observed in the HCC cells transfected with the antisense KAI1 gene. KAI1 gene transfection also affected ECM and ICAM‐1 expression in the transplanted liver cancer. Conclusion: The KAI1 gene plays an important role in the invasion and metastasis of human HCC and its upregulation in HCC cells suppresses their invasive and metastatic abilities. KAI1 gene functioned as a metastasis inhibitor by regulating the HCC cell biophysical behaviours including aggregation, adhesion, motility and visco‐elastic properties.     

The mode of tumour progression in combined hepatocellular carcinoma and cholangiocarcinoma: an immunohistochemical analysis of E‐cadherin,alpha‐catenin and beta‐catenin

Abstract: Background/Aim: To investigate the mode of progression of combined hepatocellular carcinoma and cholangiocarcinoma (cHCC‐CC). Methods: An immunohistochemical study for E‐cadherin (ECD) and alpha‐ and beta‐catenins was performed on 29 cases of cHCC‐CC. Results: Reduced expression of ECD was significantly correlated with the tumour grade of the hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC) components, intrahepatic metastasis (IM) of HCC, IM of CC, and vascular invasion of CC (p < 0.05, respectively). There was a significant relationship between the reduced expression of beta‐catenin and the tumour grade of HCC components (p < 0.05). Cases showing concurrent intrahepatic metastasis composed of HCC, CC, or both, numbered 6, 5, and 2, respectively. The expression patterns of ECD and beta‐catenin of IM were similar to those of primary lesion in most cases. On the other hand, expression of ECD and beta‐catenin of IM of HCC component were preserved, even though those of the primary sites were reduced in two cases and one case, respectively. ECD and beta‐catenin were significantly correlated with tumour differentiation and tumour progression. Conclusions: Preserved or recovered ECD and beta‐catenin expression may be of beneficial effect for re‐establishing the tissue architecture at the metastatic site.     

Melatonin‐mediated downregulation of ZNF746 suppresses bladder tumorigenesis mainly through inhibiting the AKT‐MMP‐9 signaling pathway

There still lacking effective treatment for bladder cancer. This study investigated whether melatonin (Mel) can suppress the growth and invasion of bladder cancer cells. Male C57B/L6 mice were categorized into control group (ie, subcutaneous injection of HT1197 bladder cancer cell line at the back] and treatment group [subcutaneous HT1197 cells + intraperitoneal Mel (100 mg/kg/d) from day 8 to day 21 after tumor cell injection]. In vitro Mel suppressed cell growth of four bladder cancer cell lines (ie, T24, RT4, HT1197, HT1376), cell migration in HT1197/HT1376, mitochondrial membrane potential (MMP) in T24 and colony formation in RT4 cells as well as arrested the cell cycle at G0 phase and inhibited the mitotic phase of T24 cells (all P < 0.0001). Protein expression of ZNF746 in RT4/T24 cells and protein expression phosphorylated (p)‐AKT/MMP‐2/MMP‐9 in HT1197/HT1376 cells were reduced following Mel treatment (all P < 0.001). Transfection of T24 cells with plasmid‐based shRNA (ie, ZNF746‐silencing) downregulated the protein expression of MMP‐9, cell growth, and invasion and attachment to endothelial cells but upregulated the colony formation (all P < 0.001). Mel suppressed oxidative stress and MMP but upregulated mitochondria mass in ZNF746‐silenced T24 cells, whereas these parameters exhibited a similar patter to Mel treatment in ZNF746‐silenced T24 cells (all P < 0.0001). In vivo study demonstrated that Mel treatment significantly suppressed cellular expressions of MMP‐9/MMP‐2, protein expressions of ZNF746/p‐AKT, and tumor size (all P < 0.001). Mel treatment suppressed the growth, migration, and invasion of bladder carcinoma cells through downregulating ZNF746‐regulated MMP‐9/MMP‐2 signaling.     

微血管密度与肝癌脉管侵袭关系的研究

探讨肿瘤新生微血管密度 (microvesseldensity,MVD)与原发性肝细胞癌 (hepatocellularcarcinoma ,HCC)脉管侵袭和转移的关系。采用免疫组化法 (streptavidinperoxidase ,S -P链霉素抗生物素蛋白 -过氧化酶 )对手术切除的 5 8例原发性肝细胞癌和 5 8例癌旁组织的石蜡包埋组织标本中的MVD进行了检测。在 4 0 0倍视野下选择 5个最密集区 ,记算MVD数量 ,取其均数。 (1)MVD在HCC组织中比癌旁组织明显增高 (P <0 0 1) ;(2 )HCC中有转移者及包膜不完整者 ,MVD明显高于无转移者及包膜完整者 ,(P <0 0 1) ;(3)HCC中脉管侵袭阳性组 ,MVD明显高于脉管侵袭阴性组 (P <0 0 1)。原发性肝细胞癌组织中的MVD与肿瘤的脉管侵袭和转移行为密切相关 ,可作为判定HCC转移及预后的指标     

Des-γ-carboxyl prothrombin is associated with tumor angiogenesis in hepatocellular carcinoma

Background and Aim: Hepatocellular carcinoma (HCC) is a hypervascular tumor, and angiogenesis plays an important role in its development. Previously, we demonstrated that des‐γ‐carboxyl prothrombin (DCP) promotes both cell proliferation and migration of human umbilical vein endothelial cells (HUVECs) by inducing the autophosphorylation of kinase insert domain receptor (KDR). In the present study, DCP‐associated tumor angiogenesis was assessed by comparing hypovascular and common hypervascular HCC. Methods: The solitary HCCs of 827 patients were classified into two groups according to the tumor density at the arterial phase of a dynamic computed tomography scan; the initial clinical data of patients with the hyper‐ and hypovascular types were compared. The HCC tissues from 95 tumors were analyzed by immunohistochemical staining for DCP and phosphorylated KDR, and intratumoral microvessel density (MVD) was analyzed to evaluate microvessel angiogenesis. Results: The serum DCP levels (320 ± 3532 mAU/mL) and tumor size (18.4 ± 9.0 mm) of patients with hypervascular HCC were significantly greater than those with hypovascular HCC (38.7 ± 80 mAU/mL and 14.6 ± 5.2 mm, P < 0.001). Immunohistochemical analysis revealed that the expressions of DCP and phospho‐KDR were significantly greater in hypervascular HCC (71.4% and 31.0%, respectively) than in hypovascular HCC (7.6% and 5.7%, respectively). Intratumoral MVD was significantly correlated with DCP (r = 0.48, P < 0.0001). Conclusions: des‐γ‐carboxyl prothrombin production is associated with tumor angiogenesis in HCC.     

Assessment of malignant potential of small hypervascular hepatocellular carcinoma using B‐mode ultrasonography

Aim: Evaluation of malignant potential is important to determine the treatment strategy for small hepatocellular carcinoma (HCC). The aim of the present study was to establish a method of assessing the malignant potential of small hypervascular HCC using B‐mode ultrasonography. Methods: One hundred and thirteen arterial hypervascular HCC nodules under 3 cm diagnosed by biopsy or surgical resection (20.5 ± 6.3 mm) were classified into two groups ultrasonographically: type 1 with (n = 27) and type 2 without (n = 86) a halo. Type 2 was categorized into three subgroups: type 2a, homogenous hyperechoic (n = 9); type 2b, hypoechoic with a smooth margin (n = 35); and type 2c, hypoechoic with an irregular or unclear margin (n = 42). Results: The mean diameter of type 2a nodules was significantly smaller than that of other HCC types (P < 0.05). Overall, moderately differentiated HCC was the predominant histological type, except for type 2a, all of which were well‐differentiated HCC. The percentage of poorly differentiated HCC was significantly higher in type 2c nodules (19%) than in other HCC types (P < 0.01). The percentage of Lens culinaris agglutinin‐reactive α‐fetoprotein (AFP‐L3) positivity was significantly higher in type 2c nodules (55%) than in other HCC types (P < 0.01). Classification on B‐mode ultrasonography was correlated with the histological differentiation and serum level, an indicator of a poor prognosis. Conclusion: The malignant potential of type 2a is the lowest and that of type 2c is the highest, both histologically and serologically. Assessment of the malignant potential of small, hypervascular HCC is possible by B‐mode ultrasonography.     

Expression of 8‐hydroxy‐2′‐deoxyguanosine in chronic liver disease and hepatocellular carcinoma

Abstract: Reactive oxygen species may be involved in the progression of chronic liver disease and the occurrence of hepatocellular carcinoma (HCC). To clarify whether clinicopathological findings in liver diseases are related to oxidative DNA damage, hepatic expression of the 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG) was examined in 75 liver disease patients, which included 32 chronic hepatitis (CH), 13 liver cirrhosis (LC) and 30 HCC patients. The CH patients had higher 8‐OHdG‐positive hepatocytes than LC (P<0.05). In CH and LC, the number of 8‐OHdG‐positive hepatocytes was correlated with alanine aminotransferase and asparate aminotransferase (P<0.01 and P<0.05, respectively). Of 30 HCC cases, 25 cases (83%) showed stronger immunoreactivity than non‐cancerous counterparts. The patients with poorly differentiated HCC had a larger tumor size and higher levels of AFP, and exhibited higher labeling indices of PCNA‐, TUNEL‐ and 8‐OHdG‐positive cells than those with well and moderately differentiated HCC. Our findings suggest that oxidative DNA damage is increased in association with necroinflammation in chronic liver disease and determination of 8‐OHdG is useful in assessing high‐grade malignancy in HCC.     

Osteopontin,a single marker for predicting the prognosis of patients with tumor‐node‐metastasis stage I hepatocellular carcinoma after surgical resection

Background and Aim: Osteopontin (OPN) has been linked to clinical outcomes in several solid tumors. However, it has not been fully evaluated whether OPN could be used as a single marker for the prognosis of patients with hepatocellular carcinoma (HCC), particularly in patients of the tumor‐node‐metastasis (TNM) stage I. Methods: A total of 151 patients with HCC who underwent surgical resection were enrolled, including 112 patients of the TNM stage I. OPN expression was evaluated using immunohistochemistry in the tissue microarrays derived from these patients. Immunoreactivity was classified according to the percentage and intensity of staining: negative (?), weak (+) and strong (++). The impact of OPN expression on survival of patients was analyzed. Results: In total, 65.6% (99 of 151) of HCC tissues expressed OPN. Overall survival in patients of OPN (?) group was significantly higher than those of OPN (+) or OPN (++) group (P = 0.049 and P = 0.001). Interestingly, in patients of the TNM stage I, OPN expression was correlated with the early recurrence after surgical resection (P = 0.001). Multivariate analysis showed that OPN expression was an independent prognostic factor for overall survival and disease‐free survival in patients with the TNM stage I HCC (hazard ratio, 2.272, P = 0.014 and 1.982, P = 0.037). Conclusions: These results suggest that OPN is commonly expressed in HCC and is a useful marker for predicting the prognosis of patients with the TNM stage I HCC, contributing to determining which individual patient needs adjuvant therapy to prevent the early recurrence after surgical resection.     

MMP‐9 and TIMP‐1 in the cord blood of premature infants developing BPD

We investigated matrix metalloproteinase‐9 (MMP‐9) and tissue inhibitor of metalloproteinase 1 (TIMP‐1) levels in the cord blood of 29 premature infants who were <30 weeks gestation. One, 8, and 14 infants developed severe, moderate and mild bronchopulmonary dysplasia (BPD), respectively, and 6 did not. MMP‐9 and TIMP‐1 levels in the cord blood were determined by ELISA. MMP‐9/TIMP‐1 ratios in the cord blood of infants who developed severe or moderate BPD (n = 9) were significantly higher than those who developed mild BPD or did not develop BPD (n = 20; P = 0.015). Multivariate linear regressions demonstrated that MMP‐9 levels and MMP‐9/TIMP‐1 ratios in the cord blood of the premature infants correlated with the oxygen supplementation period (r = 0.58, P = 0.003 and r = 0.41, P = 0.030, respectively). The MMP‐9 levels and MMP‐9/TIMP‐1 ratios correlated with the severity of maternal chorioamnionitis (both trend P = 0.006). The MMP‐9 levels and MMP‐9/TIMP‐1 ratios in the cord blood may be related to the pathogenesis and severity of BPD and maternal chorioamnionitis. Pediatr Pulmonol. 2009; 44:267–272. © 2009 Wiley‐Liss, Inc.     

Postoperative adjuvant transarterial chemoembolization for participants with hepatocellular carcinoma: A meta‐analysis

Aim: The efficacy of transarterial chemoembolization (TACE) for inoperable hepatocellular carcinoma (HCC) is positive, but for postoperative HCC, many studies have reported controversial results. The present study aimed to evaluate the efficacy of postoperative adjuvant TACE for participants with HCC. Methods: Electronic and manual searches were conducted to identify randomized controlled trials (RCT) evaluating postoperative adjuvant TACE for participants with HCC. Results: Six RCT totaling 659 participants, of whom almost all were of stage IIIA HCC, were included. For the 1‐year tumor recurrence rate, hepatectomy plus TACE showed statistically significant less incidence of recurrence, with a pooled risk ratio (RR) of 0.68 (95% confidence interval [CI] = 0.55–0.84, P = 0.0003). For 1‐year mortality, the trials were favorable for TACE with a pooled risk ratio of 0.48 (95% CI = 0.35–0.65, P < 0.00001). For 3‐year mortality, the trials also revealed statistically significant less incidence, with a pooled risk ratio of 0.76 (95% CI = 0.64–0.90, P = 0.002). However, for 5‐year mortality, TACE did not demonstrate statistically significant less incidence (RR = 0.94, 95% CI = 0.81–1.08, P = 0.36). Transient fever and nausea/vomiting were reported as side‐effects of TACE but were well tolerated by most participants. Conclusion: Postoperative adjuvant TACE seems promising for participants with HCC with risk factors (multiple nodules of >5 cm or vascular invasion) but requires further trial.     

Comparison between `in vivo' and `in vitro' methods for evaluating tumor angiogenesis using cervical carcinoma as a model

The role of angiogenesis in tumorigenesis is widely accepted. Therefore, it is mandatory to develop a clinically useful method for assessing tumor angiogenesis. This study was designed to compare the `in vivo' and `in vitro' methods for assessing angiogenesis and to evaluate their clinical application using cervical carcinoma as a model. Ninety women with stages IB-IIA cervical carcinoma exhibiting visible cervical tumors by transvaginal ultrasound were enrolled in this study. All patients underwent radical abdominal hysterectomy and pelvic lymph node dissection. Vascularity index (VI) was assessed by power Doppler ultrasound and a quantitative image processing system. The microvessel density (MVD) of the excised tumors was immunohistochemically assessed. Both the enzyme immunoassay and immunohistochemistry methods were performed for assessing the protein levels of vascular endothelial growth factor (VEGF) in tumor tissues. Significantly higher VI, MVD and cytosol VEGF concentrations were detected in tumors with deep stromal invasion (≥1/2 thickness) (11.43 ± 7.25 vs. 5.87 ± 6.81, P < 0.001; 53.0 vs. 37.0, P = 0.006, 550.0 vs. 86.0 pg/mg, P < 0.001), lymphatic invasion (12.21 ± 7.89 vs. 6.86 ± 6.29, P < 0.001; 53.0 vs. 40.0, P = 0.038; 930.0 vs. 110.0 pg/mg, P = 0.002), and pelvic lymph node metastasis (17.15 ± 8.58 vs. 7.83 ± 6.41, P < 0.001; 54.0 vs. 39.0, P = 0.02; 964.0 vs. 131.0 pg/mg, P = 0.002). VEGF-rich tumors detected by immunohistochemistry also revealed higher VI (12.26 ± 7.96 vs. 8.05 ± 7.62, P = 0.012), MVD (53.0 vs. 37.5, P = 0.01) and cytosol VEGF levels (745.0 vs. 98.0 pg/mg, P = 0.002). The relationships between VI values, MVD values and cytosol VEGF concentrations were linear (VI vs. MVD, r = 0.38, P < 0.001; VI vs. VEGF, r = 0.78, P < 0.001; MVD vs. VEGF, r = 0.29, P = 0.006). As revealed by the receiver operating characteristic (ROC) curve analysis, VI is better than MVD and VEGF in predicting lymph node metastasis. In conclusion, there is histological, molecular and clinical evidence supporting VI as a useful `in vivo' indicator of tumor angiogenesis, particularly for predicting lymph node metastases in cervical carcinomas. This revised version was published online in June 2006 with corrections to the Cover Date.     

Influence of miR‐520e‐mediated MAPK signalling pathway on HBV replication and regulation of hepatocellular carcinoma cells via targeting EphA2

We determined the role of miR‐520e in the replication of hepatitis B virus (HBV) and the growth of hepatocellular carcinoma (HCC) cells. MiR‐520e and EPH receptor A2 (EphA2) in HBV‐positive HCC tissues and cells were detected, and we studied the impact of miR‐520e and the EphA2 receptor in cellular and murine HBV replication models. We find that MiR‐520e was upregulated and EphA2 was downregulated in HBV‐positive HCC tissues and cells. MiR‐520e was decreased in Huh7‐X and HepG2‐X cells in which HBx was stably expressed, but was dose‐dependently elevated after interfering with HBx. Additionally, miR‐520e mimic and si‐EphA2 groups were reduced in association with increases in HBV DNA content, HBsAg and HBeAg levels, cell proliferation and were enhanced in the expressions of EphA2, p‐p38MAPK/p38MAPK, phosphorylated extracellular signal‐regulated kinase 1/2 (p‐ERK1/2)/ERK1/2 and cell apoptosis. Furthermore, si‐EphA2 reversed the promotion effect of miR‐520e inhibitor on HBV replication and tumour cell growth. Upregulating miR‐520e in rAAV8‐1.3HBV‐infected mouse resulted in reduced EphA2 in liver tissues and HBV DNA content in serum. We find that MiR‐520e was decreased in HBV‐positive HCC, while overexpression of miR‐520e blocked p38MAPK and ERK1/2 signalling pathways by an inhibitory effect on EphA2 and ultimately reduced HBV replication and inhibited tumour cell growth. These data indicate a role for miR‐520e in the regulation of HBV replication.