Predictive Factors for Differentiating Pancreatic Cancer-Associated Diabetes Mellitus from Common Type 2 Diabetes Mellitus for the Early Detection of Pancreatic Cancer.

BACKGROUND: The purpose of this study was to find the factors that may be helpful for differentiating pancreatic cancer-associated diabetes mellitus (PC + DM) from common type 2 diabetes for the early diagnosis of pancreatic cancer. METHODS: From January 2008 to August 2013, 171 patients with pancreatic cancer and new-onset diabetes were recruited for the study; 242 age- and gender-matched patients with common type 2 diabetes were also identified as control during the same period. The patient's characteristics and laboratory parameters were compared between the 2 groups. RESULTS: By multivariate logistic regression analysis, we found that body mass index (BMI), the age of onset of diabetes, hepatitis B virus (HBV) infection, total bilirubin (TBIL), alanine aminotransferase (ALT), creatinine (Cr), apolipoprotein-A1 (APO-A1), and white blood cell (WBC) were independent predictive factors for differentiating PC + DM from common type 2 diabetes; the areas under receiver operating characteristic curves were up to 0.815 for the combination of these 8 markers. CONCLUSIONS: These results suggest that BMI, the age of onset of diabetes, HBV infection, TBIL, ALT, Cr, APO-A1, and WBC are factors that could differentiate PC + DM from common type 2 -diabetes and may be used for early diagnosis of pancreatic cancer.

Prophylactic hepatic artery infusion chemotherapy improved survival after curative resection in patients with hepatocellular carcinoma.

BACKGROUND/AIMS: To explore the effect of prophylactic hepatic artery infusion chemotherapy (HAIC) on survival probability after curative resection in patients with hepatocellular carcinoma (HCC). METHODOLOGY: 85 patients with HCC were randomly assigned to HAIC group (42 cases) and control group (43 patients), all the database of two groups had no significant difference. Patients in HAIC groups underwent hepatic artery infusion chemotherapy (5-FU 1000 mg/m2 on day 1, Oxaliplatin 85 mg/m2 on day 1 and Gemcitabine 1000 mg/m2 on day 1 and 8) starting 3 weeks after operation with intervals of 4 weeks. All patients were followed up for 3 years and intrahepatic recurrence-free survival, disease-free survival rate and overall survival rate were recorded. RESULTS: Intrahepatic recurrence rate of HAIC group and the control group was respectively 19.05% and 39.53%, P < 0.05. Disease-free survival rate was respectively 57.14% and 44.19%, P < 0.05. Overall survival rate was 66.67% and 46.51%, P < 0.05. All patients in HAIC group tolerated the therapy. No adverse effect above grade 3 was reported in HAIC group. CONCLUSION: HAIC effectively and safely prevents intrahepatic recurrence and improves the prognosis of patients with HCC after curative resection.

[Three different strategies of urine drainage following hypospadias surgery: clinical nursing and observation].

OBJECTIVE: To study 3 different strategies of urine drainage following hypospadias urethroplasty, the clinical nursing in their application, and their effects. METHODS: We retrospectively analyzed the clinical data of 595 cases of hypospadias treated by urethroplasty. After surgery, 133 of the patients underwent urine drainage by suprapubic cystostomy (group A), 202 by urethral stent- tube indwelling (group B), and 260 by early initiative micturition with the urethral stent-tube (group C). All the patients received routine postoperative nursing care required for hypospadias repair. RESULTS: Operations were successfully completed in all the cases. Group C showed a remarkably shorter hospital stay and lower incidence rates of urinary fistula and urethral stricture than groups A and B (P<0.05), but there were no significant differences in the three indexes between A and B (P<0.05). CONCLUSION: For urine drainage following hypospadias repair, early initiative micturition with the urethral stent-tube can significantly reduce postoperative complications, decrease difficulties and workload of nursing care, and shorten the hospital stay of the patient.

[Clinical study of liver resection with bipolar radiofrequency device: Habib 4X].

OBJECTIVE: To assess the application value of a new radiofrequency device Habib 4X in liver resection. METHODS: A retrospective study was performed during March 2010 to July 2011.Forty-four patients underwent liver resection with radiofrequency device Habib 4X and another 54 patients traditional liver resection.Intraoperative blood loss, blood transfusion, Pringle's maneuver requirement, liver parenchyma transaction time, liver function recovery, complications, mortality and recurrence were recorded. RESULTS: The mean resection time was (67 ± 22) min for Habib 4X group versus (93 ± 23) min for traditional group (P = 0.000). Pringle's maneuver was required in 10 patients (22.7%) for Habib 4X group and 31 (57.4%) for traditional group (P = 0.001). The mean blocking time was (7 ± 2) vs (18 ± 6) min (P = 0.001), mean blood loss volume (243 ± 132) vs (500 ± 421) ml (P = 0.002). Postoperative recovery of liver function was better in Habib 4X group than traditional group. None developed mortality in Habib 4X group. And no resection margin recurred during a 18-month follow-up. CONCLUSIONS: Bipolar radiofrequency device Habib 4X is recommended for pre-coagulation in hepatectomy. And the advantages of minimized blood loss and reduced resection time result in its lower rates of morbidity and mortality.

Novel blood biomarkers of pancreatic cancer-associated diabetes mellitus identified by peripheral blood-based gene expression profiles.

OBJECTIVES: The prognosis of pancreatic cancer is still very poor, and the ability to detect pancreatic cancer in high-risk groups at an early stage is therefore essential for improving its long-term survival. The purpose of this study was to explore specific biomarkers that can differentiate pancreatic cancer-associated diabetes from type 2 diabetes, for the early detection of pancreatic cancer. METHODS: From January 2006 to July 2008, 102 peripheral blood samples were collected from 25 patients diagnosed with pancreatic cancer and diabetes, 27 patients with pancreatic cancer without diabetes, 25 patients with diabetes mellitus >5 years, and 25 healthy controls. Thirty-two samples were used in microarray experiments to find differentially expressed genes specific for pancreatic cancer-associated diabetes. The results were further validated by quantitative real-time PCR for 101 blood samples. Protein expression of selected genes in serum and tissues was also detected. RESULTS: Using microarray analysis, we found 58 genes to be unique in patients with pancreatic cancer-associated diabetes, including 23 upregulated genes and 35 downregulated genes. Eleven upregulated genes were further validated by RT-PCR, and two of these genes-vanin-1 (VNN1) and matrix metalloproteinase 9 (MMP9)-were selected for logistic regression analysis. The combination of VNN1 and MMP9 showed the best discrimination of pancreatic cancer-associated diabetes from type 2 diabetes. The protein expression of MMP9 and VNN1 was in accordance with the gene expression. CONCLUSIONS: Our results indicate that the combination of VNN1 and MMP9 may be used as a novel blood biomarker panel for the discrimination of pancreatic cancer-associated diabetes from type 2 diabetes.

The potential effect of proteasome inhibitor PS-341 on severe acute pancreatitis detected by positron emission tomography in ICR mice.

INTRODUCTION: Severe acute pancreatitis is a life threatening disease with a high rate of mortality, and its treatments are still controversial. The purpose of this study is to investigate the potential effects of proteasome inhibitor PS-341 on severe acute pancreatitis induced by cerulein and lipopolysaccharide in mice. MATERIALS AND METHODS: Severe acute pancreatitis was induced by seven intraperitoneal injections of 50 ug/kg cerulein at hourly intervals and one injection of 10mg/kg lipopolysaccharide in mice. Thirty min before the administration of lipopolysaccharide, mice were treated either with PS-341 or vehicle. The severity of acute pancreatitis was then evaluated by serum and pancreatic biochemical assays as well as histologic examination. Positron emission tomography (PET) was used for the first time to determine the therapeutic effects of interventions in situ. RESULTS: PS-341 significantly inhibited NF-kappaB activation, while the pancreatic cell apoptosis was significantly enhanced, resulting in the improved parameters such as serum amylase, C-reactive protein, lactate dehydrogenase, interleukin-1beta, interleukin-6, and pancreatic myeloperoxidase activity. Accordingly, pancreatic damage, including inflammatory cell infiltration, hemorrhage, and necrosis, was markedly reduced. (18)F-fluorodeoxyglucose-positron emission tomography demonstrated that PS-341 significantly reduced the uptake of (18)F-fluorodeoxyglucose within the pancreas. CONCLUSIONS: These observations demonstrate that PS-341 was able to significantly reduce the severity of acute pancreatitis induced by cerulein and lipopolysaccharide in mice. The potential effect is associated with the inhibition of NF-kappaB activation and increased pancreatic cell apoptosis within the pancreas. (18)F-fluorodeoxyglucose-positron emission tomography could be a sensitive and promising means in evaluating the therapeutic effect and adjusting medical interventions for pancreatitis.

Human bone marrow mesenchymal stem cells differentiate into insulin-producing cells upon microenvironmental manipulation in vitro.

It was recently reported that pluripotent mesenchymal stem cells (MSCs) in rodent bone marrow (BM) have the capacity to generate insulin-producing cells (IPCs) in vitro. However, little is known about this capacity in human BM-MSCs. We developed a nongenetic method to induce human BM-MSCs to transdifferentiate into IPCs both phenotypically and functionally. BM-MSCs from 12 human donors were sequentially cultured in specially defined conditions. Their differentiation extent toward beta-cell phenotype was evaluated systemically. Specifically, after induction human BM-MSCs formed spheroid islet-like clusters containing IPCs, which was further confirmed by dithizone (DTZ) staining and electron microscopy. These IPCs expressed multiple genes related to the development or function of pancreatic beta cells (including NKX6.1, ISL-1, Beta2/Neurod, Glut2, Pax6, nestin, PDX-1, ngn3, insulin and glucagon). The coexpression of insulin and c-peptide was observed in IPCs by immunofluorescence. Moreover, they were able to release insulin in a glucose-dependent manner and ameliorate the diabetic conditions of streptozotocin (STZ)-treated nude mice. These results indicate that human BM-MSCs might be an available candidate to overcome limitations of islet transplantation.

Pancreatic cancer-derived exosomes promoted pancreatic stellate cells recruitment by pancreatic cancer.

Cancer-associated fibroblasts (CAFs), which are an important component of the tumor microenvironment, have been identified in the blood circulation of patients with cancer metastasis, and metastatic cancer cells can recruit circulating CAFs. However, primary carcinoma sites usually regulate the behavior of metastatic cancer cells through exosomes. Here, we hypothesized that cancer-derived exosomes could enhance CAF recruitment. Exosomes secreted by pancreatic cancer cells (PANC-1 and MIA PaCa-2) were isolated and characterized. The ability of pancreatic cancer to recruit pancreatic stellate cells (PSCs) was assessed with Transwell assays in vitro and bioluminescent imaging in a mouse model in vivo, and the underlying molecular mechanism was also investigated. The results showed that pancreatic cancer cell-derived exosomes (Exo-Pan and Exo-Mia) promoted the pancreatic cancer recruitment of PSCs. This effect was mediated partially by the transfer of the exosomal protein Lin28B to the recipient cells to activate the Lin28B/let-7/HMGA2/PDGFB signaling pathway. These results suggested that exosomes derived from local cancer could promote the formation of distant metastases through transferring the exosomal protein Lin28B to the metastatic cancer cells.

Expression of tumor necrosis factor-alpha converting enzyme in liver regeneration after partial hepatectomy.

AIM: To study the expression of tumor necrosis factor-alpha converting enzyme (TACE) and evaluate its significance in liver regeneration after partial hepatectomy in vivo. METHODS: Male SD rats underwent 70% partial hepatectomy. The remaining liver and spleen tissue samples were collected at indicated time points after hepatectomy. TACE expression was investigated by Western blotting, immunohistochemistry, and serial section immunostaining. RESULTS: Expression of TACE in liver and spleen tissues after partial hepatectomy was a time-dependent alteration, reaching a maximal level between 24 and 48 h and remaining elevated for more than 168 h. TACE protein was localized to mononuclear cells (MNC), which infiltrated the liver from the spleen after hepatectomy. The kinetics of TACE expression was in accordance with the number of TACE-staining MNCs and synchronized with those of transforming growth factor-alpha (TGFalpha). In addition, TACE-staining MNC partially overlapped with CD3+ T lymphocytes. CONCLUSION: TACE may be involved in liver regeneration by pathway mediated with TGFalpha-EGFR in the cell-cycle progressive phase in vivo. TACE production and effect by paracrine may be a pathway of involvement in liver regeneration for the activated CD3+ T lymphocytes.

[Ultrasound-guided percutaneous radiofrequency ablation of hepatic malignancies with cool-tip needle].

OBJECTIVE: To investigate the efficacy, indication and complication of radiofrequency ablation (RFA) with cool-tip needle in patients with hepatic malignancies. METHODS: 421 patients with hepatic malignancies underwent ultrasound-guided RFA with cool-tip needle under local anaesthesia. The tumor size was from 1.0 to 15 cm in diameter with an average diameter of 4.3 cm. RESULTS: The complete ablation (CA) rate was 91.4% (382/418) in the patients with a tumor < 3 cm, 78.9% (97/123) in those with a tumor of 3 to 5 cm and 37.6% (35/93) in the patients with a tumor > 5 cm. No patient died or changed to celiotomy during the 1121 times of RFA for 634 lesions in 421 patients. No hemorrhage occurred in any of these patients after the RFA treatment. The complications included abdominal pain in 32.3% (136/421), nausea in 9.0% (38/421), fever in 34.9% (147/421) and biliary leakage in 0.2% (1/421) of the patients. CONCLUSION: Ultrasound-guided percutaneous radiofrequency ablation with cool-tip needle is effective and safe in the treatment of liver tumors.

[Treatment of unresectable pancreatic carcinoma by radiofrequency ablation with 'cool-tip needle': report of 18 cases].

OBJECTIVE: To explore the effects of treatment of unresectable pancreatic tumors by radiofrequency ablation (RFA) with "cool-tip needle". METHODS: 18 patients with unresectable pancreatic tumors, 8 with pancreatic head carcinoma and 10 with pancreatic body and tail carcinomas, 12 males and 6 females, aged 66.2, underwent RFA under laparotomy for 3 times and simultaneous infusion of iced normal saline. The clinical data were retrospectively analyzed. RESULTS: The level of CA19-9, a tumor marker, returned to normal after RF in 2 patients. Back pain was alleviated in 14 patients. B mode ultrasonography or CT examination showed decrease of tumor volume in 14 patients after RF. Pancreatic fistula occurred in 3 patients (16.7%) and then healed smoothly in 7 - 10 days with after routine abdominal drainage. The mortality was 22.2% (4/18). In the 4 death cases, tumors were all located in the pancreatic head; three patients died suddenly of massive gastrointestinal hemorrhage at Days 4, 30, and 40 days postoperative respectively after RF and the other patient died of acute renal failure at Day 2 postoperative days after RF. 8 patients died 1 - 8 months after RF. After 51 months, 1 patient still survived. CONCLUSION: RFA is effective in relieving the back pain for in unresectable pancreatic tumor patients efficiently relieving the back pain. Standard use of cool-tip RFA is dangerous for pancreatic head tumor carcinoma close to portal vein, but safe for those located in the body and tail of the pancreas. Making the Infusion of iced cooling-water flow via a gastric tube into the duodenum and changing change of the parameters of the RF system can reduce the associated complications.

Radiofrequency ablation versus hepatic resection for breast cancer liver metastasis: a systematic review and meta-analysis.

OBJECTIVE: To evaluate the comparative therapeutic efficacy of radiofrequency ablation (RFA) and hepatic resection (HR) for breast cancer liver metastases (BCLMs). METHODS: Studies that had examined the outcomes for both RFA and HR for BCLM were identified by searching the electronic databases PubMed, EMBASE, and the Cochrane Library. Pooled analyzes of the overall survival (OS), disease-free survival (DFS), and short-term outcomes of BCLM were performed. RESULTS: Patients with BCLM gained many more survival benefits from HR than from RFA with regard to the 3-year OS rate (combined odds ratio (OR) 0.41, 95% confidence interval (CI) 0.29-0.59, P<0.001), 5-year OS rate (combined OR 0.38, 95% CI 0.32-0.46, P<0.001), 3-year DFS (combined OR 0.36, 95% CI 0.27-0.49, P<0.001), and 5-year DFS (combined OR 0.51, 95% CI 0.40-0.66, P<0.001). RFA had fewer postoperative complications (combined OR 0.30, 95% CI 0.20-0.44, P<0.001) and shorter hospital stays (combined OR -9.01, 95% CI -13.49-4.54, P<0.001) than HR. CONCLUSIONS: HR takes precedence over RFA in the treatment of patients with BCLM, considering the better survival rate. RFA gives rise to fewer complications and can be carried out with a shorter hospital stay, compared to HR. RFA should be reserved for patients who are not optimum candidates for resection.

Risk factors for infectious morbidity in gallbladder cancer patients treated surgically.

BACKGROUND/AIMS: To explore risk factors associated with postoperative infectious morbidity in gallbladder cancer patients. METHODOLOGY: We investigated 58 consecutive patients undergoing surgery for primary gallbladder cancer between January 2000 and June 2005 in our hospital. Records of all patients were retrospectively reviewed. Twenty-two independent tumor-, patient-, and treatment-related variables were analyzed. The dependent variable was clinical infectious complications. A binary logistic regression analysis was used to assess the independent association of variables with the dependent variable. RESULTS: Overall surgical morbidity was 33% (19/58), and 14 (24%) of the 58 patients developed infectious complications postoperatively. On univariate analysis, presence of jaundice, hypoalbuminemia and intraoperative blood transfusion were found to be significantly associated with infectious morbidity. The multivariate analysis of logistic regression disclosed that presence of jaundice and intraoperative blood transfusion of 4 units or more only showed marginally significant impacts on infectious complications (odds ratio, 8.004, 7.782; 95% confidence interval, 0.886-72.278, 0.914-66.283, respectively), while weight loss and hypoalbuminemia were significantly associated with infectious complications postoperatively (odds ratio, 9.609, 40.257; 95% confidence interval, 1.269-72.253, 3.901-415.438, respectively). CONCLUSIONS: Hypoalbuminemia and weight loss are significantly associated with postoperative infectious morbidity independently. While presence of jaundice and intraoperative blood transfusion of 4 units or more appear to be marginally significant factors, modality of operation or liver resection, blood loss, and additional gastrointestinal operation are not risk factors.

Diagnosis and surgical treatment of pancreatic endocrine tumors in 36 patients: a single-center report.

BACKGROUND: Pancreatic endocrine tumors (PETs) are rare and their surgical treatment is often debated. The purpose of this retrospective study was to analyze the diagnosis and surgical strategy of functioning and non-functioning PETs. METHODS: From May 1980 to March 2006, 36 patients with pancreatic endocrine tumors at the Second Affiliated Hospital of Zhejiang University were retrospectively studied. RESULTS: Among the 36 patients, 29 (81%) had functioning tumors, and 7 (19%) had nonfunctioning tumors. Ninety-two percent of insulinomas were benign, whereas 4 (57%) of nonfunctioning PETs were malignant. The size of functioning tumors was (2.3 +/- 0.3) cm, that of nonfunctioning tumors was less than (5.1 +/- 0.5) cm. The combination CT and transabdominal ultrasonography resulted in a diagnostic sensitivity of 84%. Thirty-three primary lesions were precisely located in 32 patients (89%). Atypical tumor resection was performed for 73% of functioning tumors, while typical pancreatectomy was performed for 6 (85%) of nonfunctioning tumors. Moreover, 5 liver resections and 1 lymph node dissection were performed. During the follow-up, fifteen complications occurred in 12 (36%) patients after operation. The 5-year survival rate for patients with benign tumors was 92% compared to 50% for those with malignant tumors. Surgical cure was achieved in 95% of patients with benign insulinomas. CONCLUSIONS: Surgical strategy for PETs depends on the size and location of the tumor and the risk of malignancy. The optimal surgical procedure is key to prevent postoperative complication. Radical resection including initial and metastatic lesion may benefit patients with malignant PETs.

Enucleation of non-invasive tumors in the proximal pancreas: indications and outcomes compared with standard resections.

OBJECTIVE: The aim of this study was to evaluate the safety and efficiency of enucleation (EU) for proximal pancreatic non-invasive neoplasms. METHODS: Patients with solitary non-invasive neoplasms in the proximal pancreas from January 1998 to April 2014 at the Second Affiliated Hospital of Zhejiang University, Hangzhou, China were included. Different operations and outcomes were analyzed. RESULTS: A total of 123 patients were enrolled. Forty patients (32.5%) underwent EU including 18 patients who had tumors close to the main pancreatic duct (MPD). Sixty-one patients (49.6%) had pancreaticoduodenectomy (PD) performed and 22 (17.9%) underwent central pancreatectomy (CP). Pathological outcomes included neuroendocrine tumors, cystic lesions, and solid pseudopapillary tumors. Operation time, intra-operative blood loss, and duration of hospital stay were significantly reduced in the EU group. PD was associated with the greatest complication rate (55.7%), followed by EU (50%) and CP (40.9%), though the pancreatic fistula rate after EU was the highest (50%), especially in patients with tumors larger than 3 cm and tumors close to the MPD. EU had advantages in the preservation of pancreatic parenchyma and endocrine and exocrine function. CONCLUSIONS: EU can be carried out safely and effectively for tumors in the proximal pancreas with improved outcomes compared with standard resections, even if the tumor is larger than 3 cm and close to the MPD.

Effects of targeted CD97 immune epitopes small interference RNA on cellular biological behaviors in MDA-MB231 malignant breast cancer cell line.

Two CD97 immune epitopes, CD97EGF (epidermal growth factor domain) and CD97Stalk (stalk domain), have different distribution patterns in malignant epidermal tumors. However, little is known about the effect of CD97EGF and CD97Stalk immune epitopes in breast cancer metastasis. To explore the effects on cell proliferation, infiltration, apoptosis, and the cell cycle, we used small interfering RNA (siRNA) against CD97EGF and CD97Stalk immune epitopes to knock down CD97 in MDA-MB231 breast cancer cells. Compared with controls, CD97 knockdown caused decreased cell growth, proliferation, migration, infiltration, and altered distribution of the percentage of cells in G0/G1 and S phase. We suggest that the potential mechanism of CD97EGF and CD97Stalk immune epitopes on the biological behaviors of MDA-MB231 breast cancer cells may be related to the altered number of N-terminal glycosylation sites, which influence the stability and signaling intensity of CD97 heterodimers.

Resection of non-cystic adenocarcinoma in pancreatic body and tail.

AIM: To report the outcome of Chinese patents with non-cystic adenocarcinoma in pancreatic body and tail (NCAPBT) after resection and to discuss its surgical strategy. METHODS: Resection of NCAPBT was performed in eight Chinese patients with complete clinical-pathological data in our hospital from January 2000 to May 2004. The surgical strategy was explored by analyzing the results of these patients. RESULTS: The resection rate of NCAPBT in patients without back pain was higher than that in patients with back pain (66.67% vs 20%, 2/3 vs 1/5). The prognosis in the group receiving palliative resection was poorer than that in the group receiving curative resection. The median survival time was 12 mo in the curative resection group and 6 mo in the palliative resection group, respectively. CONCLUSION: The overall survival time of the Chinese patients with NCAPBT is dismal. The Chinese patients after curative resection of NCAPBT have a longer survival time. The Chinese NCAPBT patients with back pain trend to have a lower curative resection rate, but back pain should not be considered a contraindication for curative resection.

3D Printout Models vs. 3D-Rendered Images: Which Is Better for Preoperative Planning?

INTRODUCTION: Correct interpretation of a patient's anatomy and changes that occurs secondary to a disease process are crucial in the preoperative process to ensure optimal surgical treatment. In this study, we presented 3 different pancreatic cancer cases to surgical residents in the form of 3D-rendered images and 3D-printed models to investigate which modality resulted in the most appropriate preoperative plan. METHODS: We selected 3 cases that would require significantly different preoperative plans based on key features identifiable in the preoperative computed tomography imaging. 3D volume rendering and 3D printing were performed respectively to create 2 different training ways. A total of 30, year 1 surgical residents were randomly divided into 2 groups. Besides traditional 2D computed tomography images, residents in group A (n = 15) reviewed 3D computer models, whereas in group B, residents (n = 15) reviewed 3D-printed models. Both groups subsequently completed an examination, designed in-house, to assess the appropriateness of their preoperative plan and provide a numerical score of the quality of the surgical plan. RESULTS: Residents in group B showed significantly higher quality of the surgical plan scores compared with residents in group A (76.4 ± 10.5 vs. 66.5 ± 11.2, p = 0.018). This difference was due in large part to a significant difference in knowledge of key surgical steps (22.1 ± 2.9 vs. 17.4 ± 4.2, p = 0.004) between each group. All participants reported a high level of satisfaction with the exercise. CONCLUSION: Results from this study support our hypothesis that 3D-printed models improve the quality of surgical trainee's preoperative plans.

Overexpression of decoy receptor 3 in hepatocellular carcinoma and its association with resistance to Fas ligand-mediated apoptosis.

AIM: To characterize the expression and genomic amplification of decoy receptor 3 (DcR3) in hepatocellular carcinoma (HCC) and to evaluate the role of DcR3 in apoptosis. METHODS: We examined 48 cases of HCC for DcR3 expression by RT-PCR and DcR3 gene amplification by quantitative genomic PCR. DcR3 protein was detected by immunohistochemistry. Terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick and labeling (TUNEL) was used to identify the apoptosis cells in tissues. Primary hepatoma cell culture and MTT test were used to evaluate the protection against FasL- and chemical-induced apoptosis by DcR3 expression. RESULTS: DcR3 mRNA overexpression was detected in 60% HCC (29/48) patients. The occurrence of HCC was not associated with amplification of the gene. One sample base substitution was found in three sites as sequence in Genbank. The expression of DcR3 in HCC was the associatied with the apoptotic index (0.067+/-0.04 vs 0.209+/-0.12, P<0.01), size of mass, stage and infiltration or metastasis (41.2% vs 71.0%, 40% vs 75%, 51.8% vs 84.6%, P<0.05). DcR3 expression could be protect hepatoma cells against apoptosis induced by FasL, but not by chemicals. CONCLUSION: These data suggest that in addition to gene amplification there may be another mechanism underlying DcR3 overexpression. The effect of overexpression of DcR3 on the apoptosis of cancer cells may have direct therapeutic implications for the management of HCC.

Inhibitory effect of adeno-associated virus-mediated gene transfer of human endostatin on hepatocellular carcinoma.

AIM: To investigate the effect of adeno-associated virus-mediated gene transfer of human endostatin on the growth of hepatocellular carcinoma (HCC). METHODS: HCC cell line Hep3B was infected with recombinant adeno-associated virus containing human endostatin gene (rAAV2-hEndo). The results of transfection were detected by RT-PCR and SDS-PAGE assay. MTT assay was used to observe the effects of supernatant of transfected cells on ECV304 cell proliferation. An animal model of HCC was established by injecting Hep3B cells subcutaneously into the back of nude mice. Intratumoral injection of rAAV2-hEndo, empty virus and phosphate-buffered saline were given sequentially. Serum endostatin was determined by ELISA, the inhibitory effect of endostatin on the growth of xenograft was assessed in 3 wk. RESULTS: The results of RT-PCR and SDS-PAGE assay confirmed that rAAV2-hEndo successfully transfected Hep3B cells, and endostatin was secreted from Hep3B cells to medium. The supernatant of transfected cells markedly inhibited the proliferation of ECV304 cells (P<0.01). Intratumoral injection of rAAV2-hEndo (2 x 10(10) v.g.) led to a sustained serum endostatin level of approximately (86.71+/-5.19) ng/mL. The tumor volume and microvessel density were less in rAAV2-hEndo group than in control groups (P<0.01). CONCLUSION: Human endostatin can be stably expressed by adeno-associated virus-mediated gene transfer and effectively inhibit the growth of HCC.