Roles and Clinical Significances of ATF6, EMC6, and APAF1 in Prognosis of Pancreatic Cancer.

Background: Pancreatic cancer (PC) is prevalent among malignant tumors with poor prognosis and lacks efficient therapeutic strategies. Endoplasmic reticulum (ER) stress and apoptosis are associated with chronic inflammation and cancer progression. However, the prognostic value of ER stress-related, and apoptosis-related genes in PC remains to be further elucidated. Our study aimed at confirming the prognostic values of the ER stress-related genes, ATF6, EMC6, XBP1, and CHOP, and the apoptosis-related gene, APAF1, in PC patients. Methods: Gene Expression Profiling Interactive Analysis 2 (GEPIA2) was used to evaluate prognosis value of ATF6, EMC6, XBP1, CHOP, and APAF1 in PC. Clinical data from 69 PC patients were retrospectively analyzed. Immunohistochemistry, Western blotting, and qRT-PCR were used for the assessment of gene or protein expression. The cell counting kit-8 (CCK-8) and the Transwell invasion assays were, respectively, used for the assessment of the proliferative and invasive abilities of PC cells. The prognostic values of ATF6, XBP1, CHOP, EMC6, and APAF1 in PC patients were evaluated using Kaplan-Meier and Cox regression analyses. Results: XBP1 and CHOP expressions were not associated with PC recurrence-free survival (RFS), overall survival (OS) and disease-specific survival (DSS). ATF6 upregulation and EMC6 and APAF1 downregulations significantly correlated with the poor RFS, OS, and DSS of PC patients. ATF6 promoted PC cell proliferation and invasion, while EMC6 and APAF1 inhibited these events. Conclusion: ATF6 upregulation and EMC6 and APAF1 downregulations may be valid indicators of poor prognosis of PC patients. Moreover, ATF6, EMC6, and APAF1 may constitute potential therapeutic targets in PC patients.

Role of TGFß3-Smads-Sp1 axis in DcR3-mediated immune escape of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a leading cause of tumour-associated mortality worldwide, but no significant improvement in treating HCC has been reported with currently available systemic therapies. Immunotherapy represents a new frontier in tumour therapy. Therefore, the immunobiology of hepatocarcinoma has been under intensive investigation. Decoy receptor 3 (DcR3), a member of the tumour necrosis factor receptor (TNFR) superfamily, is an immune suppressor associated with tumourigenesis and cancer metastasis. However, little is known about the role of DcR3 in the immunobiology of hepatocarcinoma. In this study, we found that overexpression of DcR3 in HCC is mediated by the TGFß3-Smad-Sp1 signalling pathway, which directly targets DcR3 promoter regions. Moreover, overexpression of DcR3 in HCC tissues is associated with tumour invasion and metastasis and significantly promotes the differentiation and secretion of Th2 and Treg cells while inhibiting the differentiation and secretion of Th1 cells. Conversely, knockdown of DcR3 expression in HCC significantly restored the immunity of CD4+ T cells. Inhibition of DcR3 expression may provide a novel immunotherapeutic approach to restoring immunity in HCC patients.

DcR3 induces epithelial-mesenchymal transition through activation of the TGF-ß3/SMAD signaling pathway in CRC.

Decoy receptor 3 (DcR3), a novel member of the tumor necrosis factor receptor (TNFR) family, was recently reported to be associated with tumorigenesis and metastasis. However, the role of DcR3 in human colorectal cancer (CRC) has not been fully elucidated. In this study, we found that DcR3 expression was significantly higher in human colorectal cancer tissues than in paired normal tissues, and that DcR3 expression was strongly correlated with tumor invasion, lymph node metastases and poor prognoses. Moreover, DcR3 overexpression significantly enhanced CRC cell proliferation and migration in vitro and tumorigenesis in vivo. Conversely, DcR3 knockdown significantly repressed CRC cell proliferation and migration in vitro, and DcR3 deficiency also attenuated CRC tumorigenesis and metastasis in vivo. Functionally, DcR3 was essential for TGF-ß3/SMAD-mediated epithelial-mesenchymal transition (EMT) of CRC cells. Importantly, cooperation between DcR3 and TGF-ß3/SMAD-EMT signaling-related protein expression was correlated with survival and survival time in CRC patients. In conclusion, our results demonstrate that DcR3 may be a prognostic biomarker for CRC and that this receptor facilitates CRC development and metastasis by participating in TGF-ß3/SMAD-mediated EMT of CRC cells.

[Adult-to-adult living-related donor liver transplantation: report of 2 cases].

OBJECTIVE: To investigate the feasibility and safety of adult-to-adult living-related donor liver transplantation using a right lobe graft. METHODS: The clinical data of 2 cases of living-related donor liver transplantation performed between July, 2010 and November, 2010 were analyzed. RESULTS: Liver transplantation was performed using a right lobe graft including the middle hepatic vein in one case and a right lobe graft without the middle hepatic vein in the other. The ratio of graft volume to standard liver volume was 46.2% and 47.3% in the two cases, with GR/WR of 0.83 and 0.80, and donor residue liver of 42.1% and 39.5%, respectively. The donor operation lasted for 6.5 h and 5 h in the two cases with blood loss of about 200-250 ml without blood transfusion. The donors recovered uneventfully without any surgical complications, whose liver function was normal 7 days after the operation, and were discharged 14 days and 16 days after the surgery, respectively. The recipient operation lasted for 8 h and 7 h with blood loss of about 800-1000 ml. The right hepatic vein, hepatic artery, portal vein and bile duct reconstruction were performed by end-to-end anastomoses in the 2 recipients. Bile duct anastomosis stricture occurred in the first recipient 2 months after transplantation and was treated with percutaneous transhepatic cholangiography and drainage. The second recipient recovered smoothly without any complications. The recipients have so far survived 9 months and 5 months, respectively. CONCLUSION: Adult-to-adult living-related donor liver transplantation is a safe and effective option for treatment of end-stage liver diseases in the context of cadaveric liver graft shortage.

Meta-analysis of alcohol consumption and risk of extrahepatic bile system cancer.

AIM: Alcohol consumption increases the risk of liver cancer. However, there is still controversy regarding alcohol consumption and the risk of extrahepatic bile system cancer (EBSC). We performed a meta-analysis to provide an overview of the relevant studies and gain more robust estimates of the relationship between alcohol consumption and risk of EBSC. METHODS: Relevant studies published between January 1966 and October 2010 were identified by searching Medline, Embase and the Cochrane Library. Studies were selected using a priori defined criteria. The strength of the relationship between alcohol consumption and risk of EBSC was assessed by adjusted odds ratio (OR). RESULTS: A total of 113 767 participants from 10 studies (nine case-control studies and one cohort study) were identified in this meta-analysis. The studies provided adjusted overall OR estimates for drinkers versus non-/low drinkers, leading to a pooled adjusted OR of 0.82 (95% confidence interval [CI] = 0.72-0.94, P for heterogeneity = 0.194, I(2) = 27.2%). The overall adjusted OR of hospital-based studies and population-based studies were 0.80 (95% CI = 0.65-0.99, P = 0.260) and 0.79 (95% CI = 0.64-0.98, P = 0.119), respectively. For the heavy drinkers, the adjusted OR significance increased to 1.58 (95% CI = 0.97-2.57, P for heterogeneity = 0.055, I(2) = 65.4%), but it had no statistical significance. CONCLUSION: There is evidence that moderate alcohol consumption lowers the risk of EBSC compared with non-/low alcohol consumption, but not heavy alcohol consumption. Further multicenter and better controlled studies are required to confirm these findings.

[Therapeutic efficacy of Nexavar on liver cancer and its relation to the expression of Ki-67 and CD34].

OBJECTIVE: To study the therapeutic effects of Nexavar on liver cancer and its relation to the expressions of Ki-67 and CD34. METHODS: Twenty-eight patients with liver cancer were treated with Nexavar. The therapeutic efficacy of Nexavar on liver cancer was observed. Liver cancer tissues were examined for the expressions of Ki-67 and CD34 by immunohistochemistry. Microvessel density (MVD) was calculated according to the expression of CD34. RESULTS: Of 28 patients, none achieved a complete response (CR), 12 had a partial response (PR), 7 had stable disease (SD), and 9 progressive disease (PD). The efficacy of Nexavar was associated significantly with Ki-67 expression. The mean MVD count was 346.03-/+146.98 in PR patients, and 89.14-/+45.66 in PD patients. There was a significant difference in MVD between PR and PD patients. CONCLUSION: There is a better efficacy of Nexavar in treatment of liver cancer in the patients who had Ki-67-positive expression and high MVD count.

[Hepatocyte apoptosis and expression of apoptosis-regulating genes during cold preservation and reperfusion injury in rat donor liver].

OBJECTIVE: To investigate the apoptosis of hepatocytes and the expression of apoptosis-regulating genes during the donor liver ischemia and reperfusion injury in rat orthotopic liver transplantation (OLT). METHODS: Seventy-two male SD rats were randomly divided into sham operation group and transplantation group. Using Ringer's lactate solution as the perfusing and preserving solution, the grafts were preserved for 4 h before orthotopic transplantation. At 1, 6 and 24 h after the reperfusion, the recipients were sacrificed, and the serum ALT and AST levels were measured; the changes of hepatocyte apoptosis was detected by TUNEL assay, and the protein expressions of the apoptosis-regulating genes were measured by flow cytometry. RESULTS: Serum ALT and AST levels were significantly higher in transplantation group than in the control group after reperfusion. In comparison with the control group, the rats in the transplantation group showed significantly increased apoptosis index in the livers, lowered Bcl-2 levels and increased FasL levels after the transplantation, especially at 6 h after liver reperfusion (P<0.01). CONCLUSION: The donor liver ischemia and reperfusion injury can promote hepatocyte apoptosis, which may be related with the high expression of Bcl-2 gene and low expression of FasL after reperfusion injury in rats with orthotopic liver transplantation.

[Effect of immunonanoparticles loaded with adriamycin on multidrug-resistant liver cancer in nude mice].

OBJECTIVE: To observe the effect of the immunonanoparticles loaded with adriamycin in reversing multidrug resistance (MDR) in liver cancer in a nude mouse model and explore the possible mechanisms. METHODS: The cytotoxicity of adriamycin, adriamycin-loaded nanoparticles, and adriamycin-loaded immunonanoparticles was assessed in a nude mouse model bearing implant tumors of adriamycin-resistant hepatoma cell line SMMC-7721/ADM. The concentration of adriamycin in the tumor tissue was determined. RESULTS: Adriamycin-loaded immunonanoparticles showed significantly stronger cytotoxicity against the implant tumors of SMMC-7721/ADM than adriamycin-loaded nanoparticles and adriamycin. Administration of adriamycin-loaded immunonanoparticles resulted in significantly higher drug concentrations in the tumor tissue than adriamycin-loaded nanoparticles and adriamycin. CONCLUSION: Adriamycin-loaded immunonanoparticles may reverse the MDR of liver cancers in vivo probably resulting from the close binding of the particles with the tumor cells to produce a high local concentration of adriamycin in the tumors.

[Antitumor effect of nanospheres coupled with the anti-human liver cancer monoclonal antibody HAb18].

OBJECTIVE: To prepare nanospheres coupled with the anti-human liver cancer monoclonal antibody HAb18 and evaluate its immunoreactivity and antitumor effects. METHODS: The nanosphere coupled with the antibody was prepared by intermolecular cross-linking the anti-human liver cancer monoclonal antibody, HAb18, with human serum albumin nanospheres containing ADM [termed HAS(ADM)-NS] via a new hetero-bifunctional cross-linker SPDP. Condensation test and immunofluorescence assay were used to evaluate the immunoreactivity of the nanospheres, and specific binding of HAb18-HAS(ADM)-NS with liver cancer cell line SMMC-7721 was observed with optical and electron microscopes. The specific cytotoxic effects on the target cells were evaluated in vitro by MTT assay. HAb18-HAS(ADM)-NS, HAS(ADM)-NS and ADM were injected separately into nude mice bearing human liver carcinoma to evaluate the inhibitory activity of HAb18-HAS(ADM)-NS in vivo. RESULTS: The immunoreactivity of HAb18-HAS(ADM)-NS was well preserved. HAb18-HAS(ADM)-NS could bind specifically with the SMMC-7721 cells. The IC(50) of HAb18-HAS(ADM)-NS against SMMC-7721 cells was 44.6 microg/ml, lower than that of HAS(ADM)-NS (345.5 microg/ml) and ADM (365.5 microg/ml). The inhibition rate of HAb18-HAS(ADM)-NS on the growth of liver cancer xenografts was significantly higher than that of HAS(ADM)-NS and ADM (P<0.001). CONCLUSION: HAb18-HAS(ADM)-NS has immunoreactivity and can actively and specifically target the liver cancer cells. The antitumor activity of HAb18-HAS(ADM)-NS is significantly higher than that of HAS(ADM)-NS and ADM.