studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 1161A
same results. A new 179 base pair silencer position
was identified which was located at 1153 bases upstream
from which was reported previously. The p68 protein could
suppress the AFP gene expression via binding to this lesion
directly and suppress the cell proliferation. In hepatocellular
carcinoma cells p68 may suppress not only AFP production
but also cell proliferation. In the clinical therapies reduction
of AFP can become the good prediction of carcinogenesis or
reduction the mass of carcinoma. This report revealed the one
aspect of the mechanism of these relationships.
Disclosures:
The following authors have nothing to disclose: Shunsuke Nojiri, Kei Fujiwara,
Noboru Shinkai, Etsuko Iio, Takashi Joh
1955
WITHDRAWN
1956
Macrophage colony-stimulating factor receptor antagonist
inhibits progression of hepatocellular carcinoma in
vivo.
Hiroshi Kono, Yoshihiro Akazawa, Shinji Furuya, Hideki Fujii; First
Department of Surgry, University of Yamanashi, Chuo, Japan
Aim; We previously reported that macrophage colony-stimulating
factor (M-CSF) is involved in hepatocarcinogenesis by
inducing an angiogenic factor via the hepatic Mφs. Therefore,
M-CSF could be targets of therapy in hepatocellular carcinoma
(HCC). The spurpose of this study was to investigate effects of
M-CSF receptor antagonist on HCC. Materials and Method;
1; isolated mouse hepatic macrophages (Mφs) or monocytes
(Mos) were cultured with media added the different dose of
M-CSF. Production of vascular endothelial growth factor (VEGF)
was assessed. Furthermore, isolated vascular endothelial cells
(VEC) were co-cultured with or without hepatic Mφs in presense
with M-CSF (100ng/ml) for 5 days and cell proliferations were
assessed in vitro. 2; C57/BL6 mice were treated with diethyl
nitrosamine (DEN) intraperitoneally. For treatment group,
M-CSF receptor antagonist (GW2580) was treated every day.
Incidence of tumors was assessed 38 weeks after treatment.
Furthermore, angiogenesis and distribution of CD163-positive
M2 Mφs were assessed. 3; Mouse HCC cells (MH134) were
implanted to same strain C3H mice by subcutaneous injection
(1 ×10 5 /animal). Tumor progression was assessed after
3 weeks. 4; In the nude mouse, human HCC cells (HuH7)
were implanted by intra-splenic injection (1 × 10 6 /animal).
Tumor progression in the spleen was assessed after 3 weeks.
5; MH134 or HuH7 (1 × 10 4 /well) was cultured with media
containing M-CSF (100ng/ml) in the presence or absence of
GW2580 (1ng/ml) for 7 days in vitro, and cell proliferation
was assessed. Result; 1; Production of VEGF increased both in
hepatic Mφs and MOs incubated with M-CSF in dose dependent
and time dependent manner. Furthermore, the production
was significantly greater in hepatic Mφs compared with that in
Mos. Importantly, proliferation of VEC was greatest in cells cultured
with hepatic Mφs in media with M-CSF. 2; Hepatic tumors
diagnosed as hepatocellular adenoma or HCC were observed
in animals treated with DEN. In contrast, tumor incidence was
significantly reduced in DEN-treated animals with GW2580.
Enhanced angiogenesis M2Mf population observed in the
DEN-treated animals was significantly blunted by treatment of
GW2580. 3 and 4; Growth of implanted both of HCC was
significantly inhibited by GW2580 in vivo. 5; Proliferations
both of HCC were not significant difference between in cells
cultured with M-CSF and cells cultured without M-CSF. Furthermore,
GW2580 did not inhibit proliferation of these cells in
vitro. Conclusions; M-CSF receptor antagonist markedly inhibited
tumor initiation and progression. Thus, M-CSF and/or its
receptor could be a new therapeutic target for HCC. .
Disclosures:
The following authors have nothing to disclose: Hiroshi Kono, Yoshihiro Akazawa,
Shinji Furuya, Hideki Fujii
1957
Promotion of Hepatocarcinogenesis by the Endocannabinoid
Anandamide and its Receptor CB1
Ki Tae Suk 1 , Ingmar Mederacke 1 , Geum Youn Gwak 2 , Robert F.
Schwabe 1 ; 1 Columbia University, New York, NY; 2 Sungkyunkwan
University, Seoul, Korea (the Republic of)
BACKGROUND: The endocannabinoid system (ECS) exerts key
roles in the development of liver fibrosis and fatty liver, two
diseases that promote hepatocarcinogenesis. Although cannabinoids
exert potent anti-tumor effects in vitro, the role of the
ECS in carcinogenesis in vivo remains elusive. AIM: To understand
the contribution of the ECS to hepatocarcinogenesis.
METHODS: Expression endocannabinoids (EC), EC-degrading
enzymes and EC receptors, was determined by LC-MS/MS,
qPCR and immunohistochemistry in mouse and patient samples.
The contribution of ECS to diethylnitrosamine (DEN)-induced
hepatocellular carcinoma (HCC) was determined in
mice deficient in fatty acid amide hydrolase (FAAH), the main
anandamide (AEA)-degrading enzyme, and in mice deficient
for cannabinoid receptor 1 (CB1), cannabinoid receptor 2
(CB2), or transient receptor potential cation channel subfamily
V member 1 (TRPV1). RESULTS: Murine and human HCCs displayed
activation of the ECS with elevated expression of CB1
and CB2 mRNA and protein, unaltered TRPV1 expression and
only moderate alterations of EC levels. Surprisingly, FAAH-deficient
mice, which have about 10-fold increased hepatic levels
of CB1 agonist AEA, displayed a 129% (p