[Show abstract][Hide abstract] ABSTRACT: Hepatocellular carcinoma (HCC) is one of the most common and lethal malignancies worldwide. Replication factor C (RFC) plays an important role in DNA replication and checkpoint control during the cell cycle. RFC is comprised of one large subunit [replication factor C, subunit 1 (RFC1)] and four small subunits [replication factor C, subunits 2-5 (RFC2‑5)]. The role of RFC3 in the development of HCC is, as of yet, not fully understood. In the present study, western blot analysis and reverse-transcription-quantitative PCR were used to measure the expression levels of replication factor C, subunit 3 (RFC3) in HCC tissues and HCC cells. Lentivirus-mediated RFC3-specific short hairpin RNA (shRNA) was used to knock down the expression of RFC3 in HCC cells in order to examine the effects of RFC3 on HCC cell proliferation and growth. Furthermore, the expression levels of cell cycle‑related proteins were also measured in the HCC cells in which RFC3 was knocked down. Our results revealed that the expression level of RFC3 was markedly upregulated in the HCC tissues and cells. In addition, MTS and cell growth assays were used to determine the viability and proliferation of the HCC cells in which RFC3 was knocked down, and the results revealed that both cell viability and proliferation were effectively suppressed. The downregulation of RFC3 expression led to HCC cell cycle arrest in the S phase, partly by regulating the epression of cell cycle-related proteins, such as p21, p53, p57 and cyclin A. The results of the present study suggest that RFC3 plays an important role in the development of HCC, and may thus be a potential biological target in the treatment of HCC.
International Journal of Molecular Medicine 09/2015; 36(5). DOI:10.3892/ijmm.2015.2350 · 2.09 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background:
Hepatocellular carcinoma (HCC) is one of the most common and lethal malignancies in the world, and hepatitis B virus (HBV) has been well established to cause HCC. Ndc80 complex is a conserved mitotic regulator dedicated to ensuring faithful chromosome segregation and plays an important role in inducing tumor formation. However, its role in HCC caused by HBV infection remains unclear.
Immunohistochemistry (IHC), Western blot (WB), and real-time qRT-PCR were used to measure the expression of Ndc80 in HBV-related HCC tissues. Ndc80-specific short hairpin RNA (shRNA) was used to knock-down Ndc80 expression in the hepatoma cell line HeG2 and HepG2.2.15, which is stable transcribed with HBV genome. Furthermore, the effect of Ndc80 on cellular proliferation and growth were examined, respectively.
The expression level of Ndc80 was remarkably up-regulated in HBV-related HCC tissues. Down-regulation of Ndc80 expression suppressed HBV replication. With cell counting and the MTS assay, cellular proliferation and growth of Ndc80 knocking-down cell line was shown to be effectively restrained.
This study suggests that Ndc80 may play an important role in the process of HBV-related HCC, and that it may be a potential biological treatment target in the future.
Gastroentérologie Clinique et Biologique 09/2015; DOI:10.1016/j.clinre.2015.08.002 · 1.64 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To explore the risk factors of portal hypertensive gastropathy (PHG) in patients with hepatitis B associated cirrhosis and establish a Logistic regression model of noninvasive prediction.
The clinical data of 234 hospitalized patients with hepatitis B associated cirrhosis from March 2012 to March 2014 were analyzed retrospectively. The dependent variable was the occurrence of PHG while the independent variables were screened by binary Logistic analysis. Multivariate Logistic regression was used for further analysis of significant noninvasive independent variables. Logistic regression model was established and odds ratio was calculated for each factor. The accuracy, sensitivity and specificity of model were evaluated by the curve of receiver operating characteristic (ROC).
According to univariate Logistic regression, the risk factors included hepatic dysfunction, albumin (ALB), bilirubin (TB), prothrombin time (PT), platelet (PLT), white blood cell (WBC), portal vein diameter, spleen index, splenic vein diameter, diameter ratio, PLT to spleen volume ratio, esophageal varices (EV) and gastric varices (GV). Multivariate analysis showed that hepatic dysfunction (X1), TB (X2), PLT (X3) and splenic vein diameter (X4) were the major occurring factors for PHG. The established regression model was Logit P = -2.667 + 2.186X1 - 2.167X2 + 0.725X3 + 0.976X4. The accuracy of model for PHG was 79.1% with a sensitivity of 77.2% and a specificity of 80.8%.
Hepatic dysfunction, TB, PLT and splenic vein diameter are risk factors for PHG and the noninvasive predicted Logistic regression model was Logit P = -2.667 + 2.186X1 - 2.167X2 + 0.725X3 + 0.976X4.
[Show abstract][Hide abstract] ABSTRACT: Hepatitis B virus core promoter (CP) mutations can increase risk of hepatocellular carcinoma. The CP region overlaps with the HBV X (HBx) gene, which has been associated with hepatocarcinogenesis. The cyclin kinase inhibitor P53 is an important regulator of cell cycle progression. We determined whether HBx mutants that result from mutations in the CP deregulate P53.
A HBx combination (combo) mutant with changes in the CP region that corresponded to A1762T/G1764A (TA), T1753A, and T1768A was constructed and expressed in L-02 and Hep3B cells. The effects of CP mutations on expression and degradation of P53, and the effects on cell cycle progression and proliferation were analyzed.
The combo mutant decreased levels of P53 and increased cyclin D1 expression, accelerated P53 degradation in L-02 cells, accelerated cell cycle progression, and increased expression of S-phase kinase-associated protein 2 (Skp2) in L-02 and Hep3B cells. Silencing of Skp2 abrogated the effects of CP mutations on P53 expression. The kinetics of P53 expression correlated with changes in cell cycle distribution.
The HBx mutant with a combination of CP mutations can up-regulate Skp2, which then down-regulates P53 via ubiquitin-mediated proteasomal degradation, increasing the risk of hepatocellular carcinoma.
[Show abstract][Hide abstract] ABSTRACT: The molecular mechanisms of the development and progression of hepatocellular carcinoma (HCC) are poorly understood. The main objective of this study was to analyze the expression of Enabled [mammalian Ena (Mena)] protein and its clinical significance in human HCC. The Mena expression was examined at mRNA and protein levels by real-time quantitative polymerase chain reaction and Western blotting analysis in ten paired HCC tissues and the adjacent normal tissues. The expression of Mena protein in 81 specimens of HCC tissues was determined by immunohistochemistry. Associations of Mena expression with the clinicopathological features were analyzed, and prognosis of HCC patients was evaluated. The result shows the expression of Mena mRNA and protein was higher in HCC than in the adjacent normal tissues in ten paired samples. Mena was mainly accumulated in the cytoplasm of tumor cells and over-expressed in 40.74 % (33/81) patients by immunohistochemical staining. Over-expression of Mena was significantly associated with poor cellular differentiation (P = 0.025), advanced tumor stage (P = 0.003) and worse disease-free survival (DFS, P < 0.001). In addition, Mena is an independent prognostic factor for DFS in multivariate analysis (HR 2.309, 95 % CI 1.104-4.828; P = 0.026). Mena is up-regulated in HCC and associated with tumor differentiation and clinical stage. Mena may be an independent prognostic marker for DFS of HCC patients.
Medical Oncology 05/2014; 31(5):939. DOI:10.1007/s12032-014-0939-y · 2.63 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: AimsCancer is not only influenced by specific tumor cells but also by the stromal microenvironment. Upon liver damage, activated hepatic stellate cells (aHSC) become highly proliferative myofibroblast-like cells and are thought to secrete molecules that influence development of hepatocellular carcinoma (HCC). The aim of this study was to investigate the role of aHSC in the development of HCC.Methods
To assess if aHSC secreted factor(s) that promote microvascular endothelial cells (MECs) tube formation, MECs were plated with aHSC conditioned medium and tube formation analyzed by light microscopy. An established transendothelial migration assay with MECs was used to evaluate the role of aHSCs in migration and metastasis. A novel in vitro and in vivo orthotopic mouse HCC tumor model was used to investigate angiogenetic, proliferative, and metastatic activity of aHSCs.ResultsWe found that aHSCs promoted angiogenesis both in vitro and in vivo through vascular endothelial growth factor (VEGF). aHSC conditioned medium increased the ability of MECs to form tubes which was dependent upon aHSC-secreted VEGF. In addition, HCC orthogenic tumors derived from coinjection of H22 cells plus aHSCs into the hepatic lobes of mice had greater cell proliferation and vascularization, as evaluated by the presence of CD34, and VEGF expression, than tumors resulting from H22 injections alone. aHSCs also migrated from the primary tumor to sites of metastasis.Conclusion
Our findings support aHSCs playing multiple roles in HCC development and metastasis.
Hepatology Research 05/2014; 45(3). DOI:10.1111/hepr.12356 · 2.74 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Acute calculous cholecystitis is a common disease in cirrhotic patients. Laparoscopic cholecystectomy can resolve this problem but is performed based on the premise that the local inflammation must been controlled. An Initial ultrasound guided percutaneous transhepatic cholecystostomy may reduce the local inflammation and provide advantages in subsequent surgery. In this paper, we detailed our experience of treating acute severe calculous cholecystitis in patients with advanced cirrhosis by delayed laparoscopic cholecystectomy plus initiated ultrasound guided percutaneous transhepatic cholecystostomy and provided the analysis of the treatment effect. We hope this paper can provided a kind of standard procedure for this special disease; however, further prospective comparative randomized trials are needed to assess this treatment in cirrhotic patients with acute cholecystitis.
Gastroenterology Research and Practice 03/2014; 2014(3):178908. DOI:10.1155/2014/178908 · 1.75 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study aimed to retrospectively investigate the expression of the phosphate and tension homologue deleted on chromosome 10 (PTEN) protein and its prognostic role in hepatocellular carcinoma (HCC) with family aggregation resulting from hepatitis B and liver cirrhosis, which have not been established. Immunohistochemical analysis was performed to evaluate the PTEN protein expression in HCC and paired para-cancerous tissues from 79 patients with HCC caused by hepatitis B and liver cirrhosis. Of these cases, 34 represented HCC with family aggregation (HCCF group), and 45 represented HCC with no family aggregation (HCCN group). Follow-up data were collected for 3 months to 10 years and analysed for HCC recurrence, survival time and prognostic risk factors. The expression of the PTEN protein in the HCC tissue was dramatically lower in the HCCF group than in the HCCN group. The six-month, one-year and two-year overall recurrence (OR) rates of the HCCF group were significantly higher than those of the HCCN group. The one-year, two-year and five-year overall survival (OS) rates of the HCCF group were lower than those of the HCCN group. Impaired PTEN protein expression was an independent prognostic risk factor that was significantly correlated with OR and OS in HCC patients. Dramatically impaired PTEN protein expression in HCC patients with family aggregation resulting from hepatitis B and liver cirrhosis was correlated with OR and OS, and impaired PTEN expression was an independent risk factor for prognosis after radical surgery.
Experimental Biology and Medicine 07/2013; 238(8). DOI:10.1177/1535370213494654 · 2.17 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Bone marrow-derived mesenchymal stem cells (BMSCs) have been reported in many studies to reduce liver fibrosis. Apart from the paracrine mechanism by which the antifibrotic effects of BMSCs inhibit activated hepatic stellate cells (HSCs), the effects of direct interplay and juxtacrine signaling between the two cell types are poorly understood. The purpose of this study was to explore the underlying mechanisms by which BMSCs modulate the function of activated HSCs.
We show here that BMSCs directly cocultured with HSCs significantly suppressed the proliferation and α-smooth muscle actin (α-SMA) expression of HSCs. Moreover, the Notch1 and Hes1 mRNA levels and the Hes1 protein level in cocultured HSCs were evidently higher than in other models. Blocking the Notch signaling pathway with Notch1 siRNA caused the increased expression of phospho-Akt and greater cell growth of cocultured HSCs. This effect was attenuated by the PI3K inhibitor LY294002.
In conclusion, our results demonstrated that BMSCs remarkably inhibited the proliferation of HSCs through a cell-cell contact mode that was partially mediated by Notch pathway activation. In addition, the PI3K/Akt pathway is involved in HSC growth inhibition by the Notch pathway.
These findings demonstrated that BMSCs directly modulate HSCs in vitro via Notch signaling cascades. Our results may provide new insights into the treatment of hepatic fibrosis with BMSCs.
Life sciences 10/2011; 89(25-26):975-81. DOI:10.1016/j.lfs.2011.10.012 · 2.70 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Fibroblast growth factor 2 (FGF-2) and its main receptor FGFR1 have been shown to promote hepatic stellate cell (HSC) activation and proliferation. However, scant information is available on the anti-fibrogenic activity of FGFR1 inhibitors. The aim of this study was to assess the impact of a selective FGFR1 tyrosine kinase inhibitor NP603 on HSC proliferation and hepatic fibrosis. We demonstrated that rat primary HSCs secreted significant amounts of FGF-2, and its tyrosine phosphorylation of FGFR1 was attenuated by NP603. NP603 inhibited HSC activaton by measuring the expression of α-smooth muscle actin (α-SMA) and the production of type I collagen using ELISA. Furthermore, NP603 (25 μM) in vitro strongly suppressed HSC growth induced by FGF-2 (10 ng/ml) and FCS. This effect correlated with the suppression of extracellular-regulated kinase (ERK) activity and its downstream targets cyclin D1 and p21. In addition, PO NP603 (20 mg·kg(-1)·day(-1)) administration significantly decreased hepatic collagen deposition and α-SMA expression in CCl(4)-treated rats. Collectively, these studies suggest that selective blocking of the FGFR1-mediated pathway could be a promising therapeutic approach for the treatment of hepatic fibrosis.
[Show abstract][Hide abstract] ABSTRACT: Multipotent mesenchymal stromal cells (MSC) have been reported to prevent the development of liver fibrosis and have emerged as a promising strategy for cell-based therapy. However, the underlying therapeutic mechanism remains unclear. Hepatic stellate cells (SC) activation is a pivotal event in the development of liver fibrosis.
We hypothesized that MSC play an important role in regulating SC proliferation and apoptosis through paracrine mechanisms. To investigate the paracrine interactions between MSC and SC, a co-culture experimental model was developed using human MSC (hMSC) and human SC (hSC).
We demonstrate that hMSC and hSC both express nerve growth factor (NGF) receptor p75. Results acquired from transwell co-culture experiments using hSC and hMSC showed that hMSC secrete NGF, which enhances hSC apoptosis. Transcription factor nuclear factor kappa B (NF-KappaB) and B cell leukemia-xl (Bcl-xl) take part in the process.
These findings demonstrated that hMSC indirectly modulate activated hSC in vitro via NGF-mediated signaling cascades and provide a potential mechanism of how transplanted MSC are effective in treating liver fibrosis.
Life sciences 07/2009; 85(7-8):291-5. DOI:10.1016/j.lfs.2009.06.007 · 2.70 Impact Factor