Junxi Xiang

miR-451: Potential role as tumor suppressor of human hepatoma cell growth and invasion

Malignant hepatoma is the leading cause of morbidity and mortality in primary liver cancer. MicroRNAs are widely accepted to act as tumor regulators to mediate tumorigenesis. Recently, miRNA-451 (miR-451) has attracted increasing attention due to its critical roles in the development of several types of cancers. Unfortunately, its function and underlying mechanism(s) of action in hepatoma remain unclear. In this study, a significant downregulation of miR-451 was observed in hepatoma cell lines. Its overexpression by administration of miR-451 mimics decreased cell viability and promoted cell apoptosis, indicating a critical role of miR-451 in cell growth. Further mechanistic analysis suggested that miR-451 overexpression accelerated cell death in a caspase-3-dependent manner, as pretreatment with its inhibitor z-VAD-fmk notably attenuated miR-451-induced cell apoptotic rates. Moreover, miR-451 upregulation abrogated cell invasive ability, accompanied with the decrease of matrix metalloproteinase-9 (MMP-9) expression levels, which may contribute to miR-451-triggered cell apoptosis. Taken together, these results reveal a prominent role of miR-451 as a tumor suppressor regulating hepatoma cell growth and invasion in a caspase-3- and MMP-9-dependent manner. Thus, our research supports this promising therapeutic agent against hepatoma.

Hepatocyte culture in autologous decellularized spleen matrix.

abstract Background and Aims: Using decellularized scaffold to reengineer liver tissue is a promising alternative therapy for end-stage liver diseases. Though the decellularized human liver matrix is the ideal scaffold for reconstruction of the liver theoretically, the shortage of liver donors is still an obstacle for potential clinical application. Therefore, an appropriate alternative scaffold is needed. In the present study, we used a tissue engineering approach to prepare a rat decellularized spleen matrix (DSM) and evaluate the effectiveness of this DSM for primary rat hepatocytes culture. Methods: Rat decellularized spleen matrix (DSM) was prepared by perfusion of a series of detergents through spleen vasculature. DSM was characterized by residual DNA and specific extracellular matrix distribution. Thereafter, primary rat hepatocytes were cultured in the DSM in a 3-dimensional dynamic culture system, and liver cell survival and biological functions were evaluated by comparison with 3-dimensional sandwich culture and also with cultured in decellularized liver matrix (DLM). Results: Our research found that DSM did not exhibit any cellular components, but preserved the main extracellular matrix and the intact vasculature evaluated by DNA detection, histology, immunohistochemical staining, vessel corrosion cast and upright metallurgical microscope. Moreover, the method of DSM preparation procedure was relatively simple with high success rate (100%). After seeding primary hepatocytes in DSM, the cultured hepatocytes survived inside DSM with albumin synthesis and urea secretion within 10 d. Additionally, hepatocytes in dynamic culture medium had better biological functions at day 10 than that in sandwich culture. Albumin synthesis was 85.67 ± 6.34 μg/107cell/24h in dynamic culture in DSM compared to 62.43 ± 4.59 μg/107cell/24h in sandwich culture (P < 0.01) and to 87.54 ± 5.25 μg/107cell/24h in DLM culture (P > 0.05); urea release was 32.14 ± 8.62 μg/107cell/24h in dynamic culture in DSM compared to 20.47 ± 4.98 μg/107 cell/24h in sandwich culture (P < 0.05) and to 37.38 ± 7.29 μg/107cell/24h cultured in DLM (P > 0.05). Conclusion: The present study demonstrates that DSM can be prepared successfully using a tissue engineering approach. The DSM is an appropriate scaffold for primary hepatocytes culture.

Overexpression of syntenin enhances hepatoma cell proliferation and invasion: potential roles in human hepatoma.

Hepatocellular carcinoma (HCC) ranks as the third leading cause of tumor-related mortality worldwide. Recently, syntenin was found to be upregulated in several tumors and to exert pivotal roles in the development of cancer. However, its function and the underlying mechanism in HCC remain to be defined. In the present study, the elevated expression levels of syntenin mRNA and protein were detected in four HCC cell lines. Overexpression of syntenin in hepatoma HCCLM3 cells enhanced cell proliferation. Furthermore, syntenin upregulation increased epidermal growth factor receptor (EGFR) expression, which accounted for syntenin‑induced cell proliferation as precondition with EGFR siRNA clearly attenuated cell proliferation in syntenin-transfected cells. At the same time, syntenin overexpression promoted cell invasion by MMP-2, as pretreatment with anti-MMP-2 antibody blocked syntenin-induced invading cell numbers. Additionally, syntenin upregulation induced the phosphorylation of p38 MAPK contributing to the increase in MMP-2 expression, as treatment with the specific inhibitor for p38 MAPK (SB203580) clearly abrogated MMP-2 expression induced by syntenin. Collectively, our results suggest that syntenin overexpression plays a critical role in promoting the proliferation and invasion of hepatoma cells. Therefore, the present study provides new insight into how syntenin accelerates the development and progression of hepatoma, and suggests that syntenin may be a promising therapeutic agent against hepatoma.

Multiple Inflammatory Myofibroblastic Tumor of the Duodenum: Case Report and Literature Review

IntroductionInflammatory myofibroblastic tumor (IMT) is a rare low-grade malignant mesenchymal tumor, which can occur at any location, although the lung is the most commonly affected organ. It is extremely rare in the duodenum and only two cases have been reported previously. We report, to our knowledge, the first case of multiple neoplastic lesions.Case reportA 20-year-old male presented with the chief complaints of intermittent right epigastric pain, nausea and vomiting. Imaging examination, electronic gastroscopy and preoperative biopsy revealed undefined lesions in the duodenum. Pancreaticoduodenectomy was performed and diagnosis of multiple IMT was confirmed by pathological biopsy of the excised tumor. A satisfactory outcome was proved by the follow-up 1 year after curative operation.ConclusionIMT can be diagnosed by histological examination and immunohistochemical test after surgical resection. Patients can benefit from radical resection with favorable prognosis.

Using a decellularized splenic matrix as a 3D scaffold for hepatocyte cultivation in vitro: a preliminary trial.

Using a decellularized liver matrix (DLM) to reengineer liver tissue is a promising therapy for end-stage liver disease. However, the limited supply of donor organs still hampers its potential clinical application, while a xenogenic decellularized matrix may bring a risk of zoonosis and immunological rejection. Therefore, an appropriate alternative scaffold is needed. In this research, we established a decellularized splenic matrix (DSM) in a rodent model, which preserved the 3D ultrastructure, the components of the extracellular matrix (ECM) and the native vascular network. The DSM and DLM had similar components of ECM, and similar mechanical properties. Hepatocytes were seeded to the DSM and DLM for dynamic culturing up to 6 d, and distributed both in decellularized sinusoidal spaces and around the vessels. The TUNEL-positive cell percentage in a dynamic culturing decellularized splenic matrix (dDSM) was 10.7%  ±  3.6% at 3d and 25.8%  ±  5.6% at 5d, although 14.2%  ±  4.5% and 24.8%  ±  2.9%, respectively, in a dynamic culturing decellularized liver matrix (dDLM) at the same time point (p  >  0.05). Primary hepatocytes in the dDSM and dDLM expressed albumin, G6pc and Ugt1a1. The gene expression of Cyp2b1, Cyp1a2 and HNF1α in the gene transcription level revealed hepatocytes had lower gene expression levels in the dDSM compared with the dDLM at 3d, but better than those in a sandwich culture. The cumulative albumin production at 6 d of culture was 80.7   ±   9.6 μg per million cells in the dDSM and 89.6   ±   4.6 μg per million cells in the dDLM (p  >  0.05). In summary, the DSM is a promising 3D scaffold for hepatocyte cultivation in vitro.

Liver regeneration using decellularized splenic scaffold: a novel approach in tissue engineering.

BACKGROUND The potential application of decellularized liver scaffold for liver regeneration is limited by severe shortage of donor organs. Attempt of using heterograft scaffold is accompanied with high risks of zoonosis and immunological rejection. We proposed that the spleen, which procured more extensively than the liver, could be an ideal source of decellularized scaffold for liver regeneration. METHODS After harvested from donor rat, the spleen was processed by 12-hour freezing/thawing x 2 cycles, then circulation perfusion of 0.02% trypsin and 3% Triton X-100 sequentially through the splenic artery for 32 hours in total to prepare decellularized scaffold. The structure and component characteristics of the scaffold were determined by hematoxylin and eosin and immumohistochemical staining, scanning electron microscope, DNA detection, porosity measurement, biocompatibility and cytocompatibility test. Recellularization of scaffold by 5 x 10(6) bone marrow mesenchymal stem cells (BMSCs) was carried out to preliminarily evaluate the feasibility of liver regeneration by BMSCs reseeding and differentiation in decellularized splenic scaffold. RESULTS After decellularization, a translucent scaffold, which retained the gross shape of the spleen, was generated. Histological evaluation and residual DNA quantitation revealed the remaining of extracellular matrix without nucleus and cytoplasm residue. Immunohistochemical study proved the existence of collagens I, IV, fibronectin, laminin and elastin in decellularized splenic scaffold, which showed a similarity with decellularized liver. A scanning electron microscope presented the remaining three-dimensional porous structure of extracellular matrix and small blood vessels. The porosity of scaffold, aperture of 45.36 +/- 4.87 μm and pore rate of 80.14% +/- 2.99% was suitable for cell engraftment. Subcutaneous implantation of decellularized scaffold presented good histocompatibility, and recellularization of the splenic scaffold demonstrated that BMSCs could locate and survive in the decellularized matrix. CONCLUSION Considering the more extensive organ source and satisfying biocompatibility, the present study indicated that the three-dimensional decellularized splenic scaffold might have considerable potential for liver regeneration when combined with BMSCs reseeding and differentiation.

The FIB-4 score predicts postoperative short-term outcomes of hepatocellular carcinoma fulfilling the milan criteria.

BACKGROUND The fibrosis score 4 (FIB-4) score is a useful tool to determine the degree of hepatic fibrosis. Liver fibrosis and cirrhosis are well-known predictors of postoperative complications after hepatectomy. This study examined the impact of FIB-4 on postoperative short-term outcomes of patients with hepatocellular carcinoma (HCC). METHODS Three hundred and fifty patients undergoing hepatectomy for HCC between 2008 and 2013 were enrolled. The receiver operating characteristic (ROC) curve analysis was performed to determine the cutoff value of the FIB-4. Univariate and multivariate analysis was performed to identify the risk factors. The correlation of the preoperative FIB-4 value with clinicopathological parameters was examined. RESULTS Postoperative complications were observed in 202 (57.7%) patients. The optimal cutoff value of the FIB-4 was set at 2.88 and 3.85 for postoperative complications and intraoperative blood loss respectively. It was also an independent prognostic factor for postoperative complications (hazard ratio [HR], 1.202; 95% CI, 1.076-1.344; P = 0.001) and intraoperative blood loss (HR, 1.196; 95% CI, 1.091-1.343; P < 0.001) by multivariate analysis. The FIB-4 was significantly correlated with age, liver function, coagulation function, blood loss, intraoperative blood transfusion (all P < 0.05). CONCLUSION Preoperative FIB-4 is a useful index to predict postoperative outcomes in patients with HCC. The FIB-4 should be assessed routinely for hepatocellular carcinoma patients.

Collagen I promotes hepatocellular carcinoma cell proliferation by regulating integrin β1/FAK signaling pathway in nonalcoholic fatty liver

Nonalcoholic fatty liver disease (NAFLD) has become a major risk factor for hepatocellular carcinoma (HCC) worldwide. However, the underlying mechanism remains insufficiently elucidated. The expression of Collagen I, an important component of extracellular matrix (ECM), was increased during the progression from simple steatosis to NASH. The purpose of this study was to investigate the role of Collagen I in NAFLD-related HCC. To study this, the decellularized liver matrix, which preserves the pathological changes of ECM, was prepared from the human fatty liver (FLM) and human normal liver (NLM). HepG2 cells cultured in FLM had a higher proliferation rate than those in NLM. SMMC-7721 and HepG2 cells cultured on Collagen I-coated plates grew faster than those on either Collagen IV- or fibronectin-coated plates. This effect was dose-dependent and associated with elevated integrin β1 expression and activation of downstream phospho-FAK. Knocking down the expression of integrin β1 significantly decreased the proliferation of HCC cells. Additionally, an orthotopic tumor model was established in NAFLD mice at different stages. The over-expressed Collagen I in the mice liver increased the expression of integrin β1 and downstream phospho-FAK, resulting in the proliferation of HCC cells. This proliferation could be inhibited by blocking the integrin β1/FAK pathway. In summary, our study demonstrated that Collagen I promoted HCC cell proliferation by regulating the integrin β1/FAK pathway. Decellularized liver matrix can be used as a platform to three-dimensionally culture HCC cells and reproduce the impact of changed ECM on the progression of NAFLD-related HCC.

Preoperative γ-glutamyl transpeptidase to platelet ratio (GPR) is an independent prognostic factor for HBV-related hepatocellular carcinoma after curative hepatic resection.

AbstractLiver fibrosis and cirrhosis is associated with the prognosis of patients with hepatocellular carcinoma (HCC) after treatment. The &ggr;-glutamyl transpeptidase to platelet ratio (GPR) is reported to predict significant liver fibrosis and cirrhosis. The aim of this study was to investigate the predictive value of preoperative GPR on the recurrence and survival of patients with HCC who underwent curative hepatectomy.A retrospective review of demographics, medical records, and prognosis of patients with hepatitis B virus (HBV)–related HCC was performed. Overall survival (OS) and disease-free survival (DFS) were evaluated using Kaplan–Meier method, and the log-rank test was used to analyze differences in recurrence and survival. Univariate and multivariate analyses were used for significance of prognostic factor.A total of 357 patients with HBV-related HCC were included in this analysis. The preoperative GPR was associated with recurrence and survival rates, independent of HCC progression or tumor marker levels, in a multivariate analysis. OS was higher in patients with a GPR <0.84 versus ≥084 (5-year survival rates 58.6% vs. 38.5%; P < 0.001). DFS was also worse in patients with a GPR ≥0.84 than in those with GPR <0.84 (5-year recurrence rates 42.8% vs. 22.8%; P < 0.001).GPR score of ≥0.84 represents a major risk factor for the poor prognosis for HBV-related HCC after hepatic resection, and GPR served as an independent predictive factor for HBV-related HCC OS.

Effect of Static Magnetic Fields on Proliferation and Apoptosis of Gallbladder Cancer Cells: An in vitro and in vivo study

Objectives: The knowledge on how static magnetic fields (SMF) of magnetic implants affect living cells and tissues is limited while these magnetic devices are more and more frequently adopted in certain surgical approaches. Methods: In this study, we exposed cultures of human gallbladder cancer cells to SMF continuously for up to 7 days, trying to simulate the exposure pattern of surgical magnetic implants, and implanted magnetic devices into tumor-bearing nude mice for 1 month, then evaluated the effect on proliferation and apoptosis of cancer cells and tissues. Results: It showed that after SMF exposure, whereas in vitro study showed decreased cell proliferation and increased apoptosis of the gallbladder cancer cells, in vivo study showed no significant differences both on proliferation and on apoptosis. Conclusions: Our findings indicate that with current magnetic biliary anastomosis devices, the SMF from magnetic implants shows little effects on proliferative and apoptotic activities of gallbladder cancer cells in vivo, while there are still possibilities that long-term continuous SMF exposure from surgical magnetic implants may interact with living cells, probably in a feeble manner.