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EC-18, a synthetic monoacetyldiacylglyceride, inhibits hematogenous metastasis of KIGB-5 biliary cancer cell in hamster model / Myung-Hwan Kim ; Heung Moon Chang ; Tae Won Kim ; Sung Koo Lee ; Jung-Sun Park ; Young-Hoon Kim ; Tae Yoon Lee ; Se Jin Jang ; Chul-Won Suh ; Tae-Suk Lee ; Sang-Hee B Kim ; Sung-Gyu Lee 1
[요약] 1
Introduction 1
Materials and methods 2
Animals and cell lines 2
EC-18 2
T cell proliferation assay 2
Elispot assay of T cells 2
Measurement of cytokine secretion 2
Measurement of [Ca²+(이미지참조)] influx by flowcytometry 2
Cytotoxic T Lymphocytes (CTL) Assay 2
RT-PCR analysis of TLR-4 3
Real-time quantitative PCR analysis of TLR-4 3
Western blot analysis of TLR-4 3
In vivo experimental protocol 3
Statistical analysis 3
Results 3
T cell proliferation assay 3
IL-2 secretion by EC-18 treated T cells 3
Secretion of cytokines by EC-18 treated T cells 4
Measurement of [Ca²+(이미지참조)] influx of EC-18 4
Cytolytic activity of T cells from CE-18 treated hamsters 4
TLR-4 mRNA expressions of LPS stimulated biliary cancer cells with and without EC-18 4
TLR-4 protein expressions of LPS stimulated biliary cancer cells with and without EC-18 4
Pathology of metastatic biliary cancer in the lungs 5
Discussion 6
ACKNOWLEDGMENT 7
REFERENCES 7
EC-18 (monoacetyldiacylglyceride) stimulates T cell production of IL-2, IL-4, IL-12,
IFN-γ, and GM-CSF in vitro. To study the effects of these cytokines stimulated by
EC-18 on cancer cells, we applied hamster biliary cancer model, a difficult cancer
to treat. Cancer (KIGB-5) cells were given intravenously to produce hematogenous
metastatic lung lesions which were treated with EC-18 at 10, 25, and 50 mg/kg/day
respectively. The fourth group was untreated control. At 4th, 8th, and 12th week the
lungs were examined. EC-18 treated groups showed only a few microscopic lung
lesions and no evidence of metastatic lesion with highest dose whereas widespread
gross lung lesions were observed in untreated control. To investigate whether the
anti-tumor effect of EC-18 is associated with suppression of tumor cell Toll-like receptor
4 (TLR-4) expression in addition to stimulation of the immune cells, KIGB-5 cells
were exposed to LPS with or without EC-18. TLR-4 mRNA and protein expression,
measured by reverse transcriptase PCR (RT-PCR), real-time quantitative PCR and
western blot analysis, showed suppression of TLR-4 expression in KIGB-5 cells
treated with EC-18 compared with control. In conclusion, EC-18 has a significant
anti-tumor effect in this experimental model of biliary cancer suggesting potential
for clinical application to this difficult cancer.*표시는 필수 입력사항입니다.
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