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We found that EC-18 stimulated the activity of CD4 +, CD8 + cells and macrophages.ĮC-18 treated CD4 + and CD8 + cells in vitro stimulate the production of cytokines IL-2, IL-4, IL-12, IFN-γ, and GMCSF ( Fig. In this paper, we showed that EC-18 has a potent anti-tumor activity in experizmental hematogenous metastatic biliary cancer in Syrian golden hamsters. We previously reported that the water extracts from deer antler, Cervus nippon, had a stimulating effects on hematopoiesis in vitro and in vivo ( 4, 5), and we subsequently demonstrated that monoacetyldiglyceride (EC-18) was the component with the most potent stimulatory activity on the bone marrow stem cells, bone marrow stromal cells and immune cells. This suppression of metastasis is likely through stimultion of immune cells (T & B cells, DC & macrophages) in association with suppression of tumor cell TLR-4 expression. We report here the use of a novel agent of EC-18 as metastasis inhibiting agent in hamster biliary cancer model (KIGB-5). EC-18 has been shown to have inhibitory effects on TLR-4 expression of experimental biliary cancer cells (KIGB-5). Activation of TLR-4 in tumor cells was found to stimulate cancer cell proliferation, whereas inhibition of TLR-4 signaling retarded tumor growth and prolonged the survival of tumor bearing mice ( 7). Toll-like receptors (TLRs), especially TLR-4, has been shown to be important in both innate and adaptive immunity to various cancers. EC-18 was an effective agent as in inhibiting the growth of metastatic biliary cancer. Using the Syrian golden hamster model of biliary cancer, we observed previously that IL-2 encoded bone marrow stromal cells (ad/hIL-2 BMSC) suppressed metastatic tumor ( 6).
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We have shown that chemically synthesized EC-18 can stimulate the proliferation of hematopoietic stem cells, bone marrow stromal cells ( 4, 5), and immune system cells, including T and B lymphocytes, dendritic cells (DCs) and macrophages, both in vivo and in vitro.
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High dose Interleukin (IL)-2 has been tested against various malignant tumors, both in experimental models and in clinical studies, but its effectiveness has been hampered by undesirable side effects ( 3).ĮC-18 is a monoacetyldiglyceride (EC-18 specifically, 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol) that occurs naturally in various seed oils, in bovine udder and in milk fat, and has been isolated from the antlers of sika deer ( Cervus nippon TEMMENICK) these antlers have been used as a tonics in oriental medicine. A variety of biological response modifiers, including cytokines and bacterial products, have been employed in immunochemotherapy of various cancers. Chemotherapy and radiation therapy have been of limited value ( 1). Surgery still remains the only curative treatment, but many of these tumors are not resectable at the time of diagnosis. and Europe over the past three decades ( 1, 2). Although it occurs with greater frequency in Asian countries, its incidence has been steadily increased in U.S.A. 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.Ĭholangiocarcinoma is a malignancy arising from the epithelial cells of the bile duct or gallbladder. 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. 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. 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. At 4th, 8th, and 12th week the lungs were examined. 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. 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.
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EC-18 (monoacetyldiacylglyceride) stimulates T cell production of IL-2, IL-4, IL-12, IFN-γ, and GM-CSF in vitro.