u-0126 and Gallbladder-Neoplasms

Primary cancer of the gallbladder is not unusual. Most cases of gallbladder cancer are found at an advanced stage, accompanied by the invasion to the liver, metastases to the lymph nodes and distant organs, and peritoneal dissemination. In this study, we first examined the effect of mitogen-activated protein kinase kinase (MEK) inhibitors on the production of matrix metalloproteinases (MMPs), urokinase-type plasminogen activator (uPA), and tissue inhibitors of metalloproteinases (TIMPs) in a human gallbladder cancer cell line, NOZ cells in vitro. MEK inhibitors (PD98059 and U0126) inhibited the production of MMP-2, MMP-9 and high MW uPA, and upregulated TIMPs (TIMP-1, TIMP-2 and TIMP-3). Subsequently, we examined the effect of U0126 on invasion and metastasis of orthotopically inoculated NOZ cells in nude mice. Direct liver invasion by cancer cells was detected in all of the mice in the control group, but in only one mouse in the U0126-treated group. Most of the primary tumors in the U0126-treated group expanded to the liver, but did not invade into the liver. Vessel invasion in the liver was evident in 4 out of 5 mice in the control group, but in only one mouse in the U0126-treated group. Lymph node metastases and peritoneal dissemination were recognized in all of the mice in both groups. All 5 mice in the U0126-treated group, and 4 out of 5 mice in the vehicle control group, had metastases in the lungs. The present results suggest that a MEK inhibitor, U0126, prolonged the survival of the mice with NOZ tumor by inhibiting direct liver invasion and vessel invasion of the cancer cells via down-regulation of the matrix degrading ability of the cancer cells.

Topics: Animals; Butadienes; Cell Line, Tumor; Cell Survival; Enzyme Inhibitors; Flavonoids; Gallbladder Neoplasms; Humans; Liver; Liver Neoplasms; Lymphatic Metastasis; MAP Kinase Kinase 1; Mice; Mice, Nude; Models, Anatomic; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplasm Transplantation; Nitriles; Urokinase-Type Plasminogen Activator

Most cases of gallbladder cancer are found at an advanced stage accompanied by invasion to the liver, metastases to the lymph nodes and distant organs, and peritoneal dissemination. Then, the treatment for advanced gallbladder cancer is often ineffective, and the prognosis of this disease is poor. The specific aim of this study was to establish a model system for developing new therapeutic strategies, such as molecular target therapy, for human advanced gallbladder cancer. We used a human gallbladder cancer cell line, NOZ with K-ras mutation in this experiment. Then, we established a novel orthotopic inoculation model for gallbladder cancer by using NOZ cells in nude mice. Mitogen-activated protein kinase (MAPK) in NOZ cells was constitutively activated, and the activation of MAPK provided autonomous proliferation of NOZ cells. All of the mice orthotopically inoculated by NOZ cells developed tumors at the gallbladder. Direct invasion into the liver, and bloody ascites were observed. Metastases to the mediastinal lymph nodes were also recognized in all of the mice examined. Moreover, most of the mice showed lung metastases. Survival duration was 29-50 days after inoculation. Nude mice with NOZ tumor at gallbladder were treated by an intraperitoneal injection of a MAPK kinase inhibitor U0126 (25 micro mole/kg) twice a week. U0126 (p=0.0110, one-way ANOVA) significantly prolonged the survival duration of the mice with NOZ gallbladder tumor. Our orthotopic inoculation model is useful for developing new therapeutic strategies, such as molecular target therapy for advanced gallbladder cancer.

Topics: Animals; Blotting, Western; Butadienes; Cell Division; Cell Line, Tumor; Enzyme Activation; Enzyme Inhibitors; Gallbladder; Gallbladder Neoplasms; Genes, ras; Humans; Immunohistochemistry; Liver; Mice; Mice, Nude; Mitogen-Activated Protein Kinases; Mutation; Neoplasm Metastasis; Neoplasm Transplantation; Nitriles; RNA; Transcription, Genetic; Tumor Suppressor Protein p53