Minoru Yamamoto

Enhanced angiogenesis due to inflammatory cytokines from pancreatic cancer cell lines and relation to metastatic potential.

Objectives: To investigate the mechanisms of metastasis formation in human pancreatic carcinoma, we examined the angiogenic capabilities of human pancreatic cancer cell lines with different metastatic potentials and the roles of inflammatory cytokines. Methods: Interleukin (IL)-8 secretion by human pancreatic cancer cells stimulated with IL-1&agr; or IL-1 receptor antagonist (IL-1ra) was measured by enzyme-linked immunosorbent assay (ELISA). We then examined how cancer cells with different metastatic potentials influenced the proliferation and tube formation of human umbilical vein endothelial cells (HUVECs) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye reduction method (MTT assay) and an angiogenesis assay, respectively. We also examined the role of inflammatory cytokines in relation to tumor metastatic potential and angiogenesis. Results: IL-8 secretion levels by pancreatic cancer cells were regulated by IL-1&agr; and correlated with metastatic potential. Both HUVEC proliferation and tube formation were strongly enhanced by coculture with metastatic pancreatic cancer cells and were enhanced to a similar extent by culture in the presence of IL-1&agr; and IL-8. In contrast, blockade of IL-1&agr; or IL-8 inhibited HUVEC proliferation and angiogenesis. Conclusions: The inflammatory cytokines IL-1&agr; and IL-8 may have an important role in metastasis via vascular endothelial cell proliferation and angiogenesis.

Interleukin-1α Enhances Integrin α6β1 Expression and Metastatic Capability of Human Pancreatic Cancer

Objective: To investigate the mechanisms of metastasis formation in metastatic human pancreatic cancer, we examined the enhancement in integrin expression, and adherence to and invasiveness into extracellular matrix (ECM) proteins of human pancreatic cancer cells after exposure to interleukin (IL)-1α. Methods: Expression of IL-1 receptor type I (IL-1RI) and alterations in integrin subunits by IL-1α were examined by flow-cytometric analysis and by cellular enzyme-linked immunosorbent assay in four human pancreatic cancer cell lines (BxPC-3, PaCa-2, PANC-1, and SW1990), respectively. In addition, assays of cancer cell adhesion and invasion to ECM proteins were performed to investigate whether increased integrin expression affected the adhesive and invasive interaction between cancer cells and the putative integrin ECM ligands. Furthermore, immunohistochemistry was used to assess integrins and IL-1R1 expression in pancreatic tissues. Results: In metastatic cancer cells, expression of the α6 subunit was enhanced by IL-1α treatment. While metastatic cancer cells exhibited preferential adherence to and invasion into laminin, these properties were enhanced by IL-1α. The α6 subunit and IL-1RI were strongly expressed in pancreatic tissues from pancreatic cancer patients with liver metastasis. Conclusions: In pancreatic cancer, IL-1α enhanced α6β1-integrin expression, probably via increased IL-1RI levels. Our results indicated that α6β1-integrin and IL-1RI expression may play important roles in metastasis formation.

Alteration of integrins by interleukin-1α in human pancreatic cancer cells

Introduction Adhesion of tumor cells to extracellular matrix (ECM) proteins plays an important role in tumor invasion and metastasis. Aims To investigate the expression of integrins in human pancreatic cancer cell lines and its alteration by interleukin (IL)-1&agr; to examine the mechanism of adhesion of metastatic human pancreatic cancer cells to ECM proteins. Methodology The expression of integrin subunits and their alteration by IL-1&agr; were examined by flow-cytometric analysis and cellular enzyme-linked immunosorbent assay in three metastatic human pancreatic cancer cell lines (AsPC-1, BxPC-3, and SW1990) and two nonmetastatic cancer cell lines (PaCa-2 and PANC-1). In addition, assays of cancer cell adhesion to ECM proteins were performed to investigate if increased integrin expression actually affected the adhesive interaction between cancer cells and the putative integrin ECM ligands. Results The &agr; 6 subunit expressed in metastatic cancer cells was enhanced by IL-1&agr;. Metastatic cancer cells also showed preferential adherence to laminin compared with nonmetastatic cancer cells, and this was enhanced by IL-1&agr;. Conclusion In pancreatic cancer, the enhancement of &agr; 6 &bgr; 1 integrin by IL-1&agr; through IL-1 receptor type I, as well as the expression of &agr; 6 &bgr; 1 integrin, plays an important role in metastasis formation.

Expression and Prognostic Roles of Integrins and Interleukin-1 Receptor Type I in Patients with Ductal Adenocarcinoma of the Pancreas

To Investigate the prognostic indicator, we examined the expression of α6- and α5- integrin and interleukin-1 receptor type I (IL-1RI) immunohistochemically, and analyzed the correlation between immunohistochemical findings and clinicopathological factors in pancreatic cancer. In patients with a strongly expressing α6- integrin subunit or weakly expressing α5β1-integrin in pancreatic cancer tissues there was a significant association with advanced TNM stage (P = 0.027 and 0.014, respectively), presence of liver metastases (P = 0.032 and 0.002, respectively), and poor prognosis (P = 0.0155 and 0.0056, respectively). In patients with a weakly expressing α6 integrin subunit or weakly expressing α5β1-integrin in noncancerous pancreatic tissues there was a significant association with poor prognosis (P = 0.0324 and 0.0396, respectively). Multivariate analysis demonstrated that strong expression of α6- and weak expression of α5β1-integrin were found to be independent prognosticators in pancreatic cancer patients. Our present results indicate that α6β1- and α5β1-integrin expression can be a significant prognostic indicator in pancreatic cancer.

Laparoscopic Management of Small Bowel Incarceration Caused by a Broad Ligament Defect: Report of a Case

We report the case of a 94-year-old woman who presented with signs of a small bowel obstruction many years after an appendectomy. Abdominal computed tomography (CT) scan showed discontinuity of the small bowel at a point next to the uterus. We made a provisional diagnosis of an internal hernia through a defect in the broad ligament and performed laparoscopic exploration, which revealed a viable ileal loop incarcerated through the broad ligament. Thus, CT scan may be useful for diagnosing this type of defect preoperatively, whereby open surgery can be avoided.

Effect of galanin on plasma glucose, insulin and pancreatic glucagon in dogs

The effect of synthetic galanin on plasma glucose, insulin and pancreatic glucagon levels in dogs was studied. Infusion of galanin caused a rapid, reversible and dose-dependent reduction in basal insulin level. A maximal increase in blood glucose level accompanying the insulin decrease was observed when galanin was administered at a dose of 4 μg/kg per h. Pancreatic glucagon levels showed little change compared with basal secretion. These results indicate that galanin is involved in the regulation of glucose through control of insulin secretion.

Glial Cell Line-Derived Neurotrophic Factor Enhances Nuclear Factor-κB Activity and Invasive Potential in Human Pancreatic Cancer Cells

Objectives: The invasive potential is increased by glial cell line–derived neurotrophic factor (GDNF) in human pancreatic cancer cell lines. We researched whether the signaling pathway activated by GDNF correlates with the nuclear factor-κB (NF-κB) in human pancreatic cancer cell lines and whether the inhibition of NF-κB activity is associated with suppression of invasive potential. Methods: Proliferation of human pancreatic cancer cell lines (BxPC-3 and MIA PaCa-2) was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays (MTT assay). NF-κB activity was examined by dual luciferase assay and electrophoretic mobility shift assay. In addition, to investigate the invasive potential, an in vitro invasion assay was performed. Results: Proliferation of both cell lines was decreased by a protea-some inhibitor, MG132, in a dose-dependent manner, but proliferation of control and IκBαM vector–transfected BxPC-3 cells was similar. The invasion cell number and the NF-κB activity were increased by GDNF stimulation. However, in the presence of MG132 or IκBαM, which blocks the nuclear localization of NF-κB, both were significantly suppressed. Furthermore, reduced activity of both remained unchanged by GDNF stimulation. Conclusion: These results indicate that GDNF promotes NF-κB activation and that the latter is involved in the invasive potential of human pancreatic cancer cells.

Synthetic Serine Protease Inhibitor, Gabexate Mesilate, Prevents Nuclear Factor-κB Activation and Increases TNF-α-Mediated Apoptosis in Human Pancreatic Cancer Cells

Gabexate mesilate (GM), a synthetic serine protease inhibitor, suppresses nuclear factor-κB (NF-κB) activity in human monocytes or human umbilical vein endothelial cells (HUVECs). In this study we examine whether GM also suppresses NF-κB activation and induces apoptosis in human pancreatic cancer cell lines. The addition of tumor necrosis factor α (TNF-α) did not change the rates of growth of BxPC-3 and MIA PaCa-2. However, in the presence of GM and TNF-α, proliferation decreased in a dose-dependent manner. GM- and TNF-α-treated cells exhibited morphologic changes indicative of apoptosis, including chromatin condensation and nuclear fragmentation. The NF-κB activity of both cell lines was increased by the addition of TNF-α, while TNF-α-induced NF-κB activity was suppressed by prestimulation with GM in a dose-dependent manner. Caspase 3 and 7 activity was significantly increased by TNF-α with GM stimulation. Furthermore, GM also suppressed the invasive potential of both cell lines. These results indicate that GM inhibits TNF-α-induced NF-κB activation and enhances apoptosis in human pancreatic cancer cell lines.

Effects of whisky on plasma gastrin and cholecystokinin in young adult men.

Whisky (1 g/kg, 21.5% alcohol) was administered orally to healthy young adult male volunteers, and changes in the plasma concentrations of alcohol, acetaldehyde, gastrin, cholecystokinin (CCK) and serum amylase were measured over time. Values for alcohol and acetaldehyde rapidly reached a peak at 30–45 min after alcohol intake, followed by a gradual decline. The plasma gastrin concentration showed a rapid elevation, while the plasma CCK concentration did not exhibit any significant changes in the early phase after alcohol intake. Elevation of CCK was observed after 75 min, however. These results show that intake of whisky stimulates the secretion of gastrin and is associated with a later increase in CCK.

Combined Gemcitabine and α-Interferon Therapy for Pancreatic Cancer: Report of a Case

Pancreatic cancer is a malignancy with an extremely poor prognosis and its incidence has increased in many countries (1–3). Less than 5% of patients with pancreatic adenocarcinomas survive for more than 5 years. The poor response of advanced pancreatic cancer is attributable to the infiltrative property of pancreatic cancer cells (4), to poor therapeutic options, and to the current incomplete knowledge concerning the pathogenesis and biology of the tumor (3). The current methods of treating pancreatic cancer are far from satisfactory, and not all patients benefit from these methods of treatment; the overall results of systemic and regional chemotherapy for pancreatic cancer are disappointing. To date, the most effective chemotherapy drug has been gemcitabine (difluorodeoxycytidine), which produces a response rate of 4–36% in pancreatic cancer patients (5–7). However, despite these relatively high response rates, a complete response is achieved in very few patients. Therefore, it is unclear whether chemotherapy actually prolongs survival. In this report, we describe the case of a patient in whom effective results were achieved using a new combination of the chemotherapeutic agents, gemcitabine and α-interferon (α-INF).