Retinoids, better known as vitamin A, have been reported to inhibit the growth of several breast cancer cell lines in culture and to reduce breast tumor growth in animal models. Furthermore, retinoids can augment the action of other breast cancer cell growth inhibitors both in vitro and in vivo. Clinically, interest has increased in the potential use of retinoids for the prevention and treatment of human breast cancer. We have examine the effect of all-trans retinoic acid(tRA) and 9-cis retinoic acid(9-cis RA) on human breast cancer cell(MCF-10A, T47-D, MCF-7) proliferation using MTT assay and cell cycle analysis(FACS). Overexpression of cyclin D1 protein is observed in the majority of breast cancers, suggesting that dysregulated expression of cyclin D1 might be a critical event in breast cancer carcinogenesis. We investigated whether tRA and 9-cis RA might affect expression of cyclin D1 on human breast cancer cells(MCF-10A, T47-D, MCF-7) using RT-PCR and west-ern bolt. In MCF-10A cells, either tRA or 9-cis RA treatment did not affect the cell proliferation. In T47-D cells and MCF-7 cells, either tRA or 9-cis RA treatment showed the inhibition of the cell proliferation over control cells and also inhibit the estrogen stimulated cell proliferation when it was given together with estrogen. The effect of retinoids was dose- and time- dependent. T47-D cells treated with 1.0 $\muM$ tRA undergo G0/G1-phase arrest by Day 5. MCF-7 cells treated with 1.0 $\muM$ tRA undergo S-phase arrest by Day 5. All-trans retinoic acid(tRA) and 9-cis retinoic acid(9-cis RA) inhibited the cyelin D1 mRNA and protein expression levels of human MCF-7 and T47-D breast carcinoma cells in vitro. The data indicate that retinoids can reduce cyclin D1 expression levels in a variety of breast cell lines in vitro and result in inhibition of cell proliferation. tRA-mediated growth inhibition and cyclin D1 expression inhibition is more potent than 9-cis RA mediated that. tRA-mediated inhibition effect is more potent on T47-D cells than on MCF-7 cells. Our data suggest that retinoids activity is different according to property of cell lines. Future chemoprevention of breast cancer studies using retinoids will be necessary to determine the mechanism of the retinoids-mediated growth inhibition.
Journal of Physiology & Pathology in Korean Medicine
/
v.21
no.5
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pp.1278-1284
/
2007
Solanum Iyratum Thunb (Solanaceae) has multiple applications in korean traditional medicine because of its cytotoxic, immunological and anti-inflammatory activities. However, no study on the anti-arthritic activity of Solanum Iyratum Thunb has been reported in vivo. Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which ultimately leads to the destruction of cartilage and bone. Cytokine production were assessed during CIA(collagen-induced arthritis) model mice in lymph node (LN), in knee joint and spleen, using ELISA. DBA/1j mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with Solanum Iyratum Thunb (SLT) orally at 400, 200 mg/kg once a day for 4 weeks. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. SLT significantly suppressed the progression of CIA and inhibited the production of TNF-alpha and IFN-gamma in serum and spleen cell culture supernatant. The erosion of cartilage was dramatically reduced in mouse knees after treatment with SLT. In conclusion, our results demonstrates that SLT significantly suppressed the progression of CIA. This action was characterized by suppression of arthritis index, cartilage erosion and synovial cell infiltration and the decreased production of $TNF-{\alpha}$, $IFN-{\gamma}$, CD4+, CD19+, CD3+/CD69+ cells (in lymph node), CD11b+/Gr-1 + (in knee joint).
Kim, Dong-Ju;Ryu, Su-Noh;Han, Sang-Jun;Kim, Hwa-Young;Kim, Jung-Hak;Hong, Seong-Gil
The Korean Journal of Food And Nutrition
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v.24
no.3
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pp.273-281
/
2011
Rice bran is byproducts of the hulling of rice, an important food resource in Korea. Various studies have been reported immune-enhancing effects of rice bran cultured with Lentinus edodes. In particular black rice bran contains anthocyanin, and the effects of antioxidant have been reported. The objective of the this study was to investigate the possible immune-enhancing effects of black rice bran substance extracted from a submerged culture of Lentinus edodes with black rice bran (crude fermentation-polysaccharide, CFP) and products(crude fermentation-polysaccharide-S. cerevisiae CFP-S, crude fermentation-polysaccharide-L. gasseri, CFP-L) which are of secondary fermentation of by using Saccharomyces cerevisiae and Lactobacillus gasseri in the Blab/c male mice. We found that supplementation of CFP, CFP-S and CFP-L enhanced macrophage and splenocyte proliferation compared to the control group(NC) in mice. Also, we measured the concentration of cytokines(IFN-${\gamma}$, TNF-${\alpha}$, IL-6) secreted by activated macrophage and splenocyte. The results of the experiment are that supplementation of CFP and CFP-S increased the macrophage and splenocyte proliferation compared to the control group but supplementation of CFP-L decreased the splenoyte proliferation compared to the control group(without mitogen and treated with LPS). When macrophage and splenocyte were stimulated by CFP and CFP-S supplementation, it was increased IFN-${\gamma}$, TNF-${\alpha}$ and IL-6 concentration compared with the control group. These results suggest that the capacity of CFP and CFP-S seem to act as a potent immune modulator causing augmentation of immune cell activity, and enhance the immue function through regulating cytokine production capacity by activated macrophage and splenocyte in mice.
Two kinds of Korean rice-wine (Yakju) with different process and ingredients, and Japanese rice-wine (Sake) were chosen for this study, and throughly dried and solubilized in water or cell culture medium. In vitro cytotoxicity assays of the solubilized wine solids exhibited that maximum dilution factors for inhibition of B 16BL6 mouse melanoma cell growth were 16X for herbal medicine-added rice-wine (Korean rice-wine I) and typical Korean rice-wine (Korean rice-wine II), and 8X for Japanese rice-wine. Their cytotoxic effects on HRT18 human colon adenocarcinoma cells were even lower than those on B16BL6 cells. The morphology of the tumor cells were changed by addition of the solubilized wine solids. Inhibitory effect of the rice-wine on in vivo tumor growth and metastasis were monitored after implantation of B16BL6 cells into C57BL/6 mice with daily feeding the solubilized wine solids. Compared to non-fed control groups, B16BL6 tumor growth and metastasis to lung were clearly inhibited by feeding the wine solids, in order of Korean rice-wine I > Korean rice-wine II > Japanese rice-wine. The data of in vitro cytotoxicity and the cell shape changes indicate that the inhibitory effect of tumor progression may be attributed to tumor cell differentiation or immune stimulation induced by certain components in the rice-wine, rather than direct cytotoxicity of the components.
Polyorach, S.;Poungchompu, O.;Wanapat, M.;Kang, S.;Cherdthong, A.
Asian-Australasian Journal of Animal Sciences
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v.29
no.9
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pp.1273-1279
/
2016
The objectives of this study were to determine an optimal cultivation time for populations of yeast and lactic acid bacteria (LAB) co-cultured in fermented milk and effects of soybean meal fermented milk (SBMFM) supplementation on rumen degradability in beef cattle using nylon bag technique. The study on an optimal cultivation time for yeast and LAB growth in fermented milk was determined at 0, 4, 8, 24, 48, 72, and 96 h post-cultivation. After fermenting for 4 days, an optimal cultivation time of yeast and LAB in fermented milk was selected and used for making the SBMFM product to study nylon bag technique. Two ruminal fistulated beef cattle ($410{\pm}10kg$) were used to study on the effect of SBMFM supplementation (0%, 3%, and 5% of total concentrate substrate) on rumen degradability using in situ method at incubation times of 0, 2, 4, 6, 12, 24, 48, and 72 h according to a Completely randomized design. The results revealed that the highest yeast and LAB population culture in fermented milk was found at 72 h-post cultivation. From in situ study, the soluble fractions at time zero (a), potential degradability (a+b) and effective degradability of dry matter (EDDM) linearly (p<0.01) increased with the increasing supplemental levels and the highest was in the 5% SBMFM supplemented group. However, there was no effect of SBMFM supplement on insoluble degradability fractions (b) and rate of degradation (c). In conclusion, the optimal fermented time for fermented milk with yeast and LAB was at 72 h-post cultivation and supplementation of SBMFM at 5% of total concentrate substrate could improve rumen degradability of beef cattle. However, further research on effect of SBMFM on rumen ecology and production performance in meat and milk should be conducted using in vivo both digestion and feeding trials.
Journal of the Korean Society of Food Science and Nutrition
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v.33
no.2
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pp.455-462
/
2004
Prostate cancer is a leading cause of cancer death in American men and evidences point to significant life style/diet components as risk factors for its development or prevention. Two large cohort studies have identified the consumption of tomatoes or high Plasma levels of Iycopene as associated with reduced risk. A number of other substances such as quercetin, phytoene, phytofluene, cyclolycopene, salicylates and tomatine in tomato besides lycopene could have anticancer activity and may be acting synergistically with lycopene. Lycopene at almost physiologically feasible concentrations, reduces cell viability by cell cycle arrest and apoptosis and modulates the cyclin pathways as well as increasing intercellular communication. However, it is not clear whether lycopene or its oxidation products are more bioactive. Tomato product supplementation results in plasma accumulation of phytoene, Phytofluene, the lycopene oxidation product, and cyclolycopene at significant concentrations and lycopene supplementation, either as a tomato product or as beadlets, results in maximal mean plasma lycopene concentrations of ∼ 1 $\mu$M which is at the lower limit of its activity in cell culture. Rats and mice are poor accumulators of lycopene and other carotenoids making them poor models for the study of cancer prevention and control. Of the 19 animal studies for various cancer sites, lycopene showed a positive effect in 10 studies but negative in 2 prostate cancer studies. In vivo prevention of leukocyte DNA damage in humans has been mostly studied using tomato product supplementation but lycopene supplementation appeared to reduce oxidative DNA damage as well as tomato product supplementation. Lycopene appears to be bioactive in intefering with carcinogenesis but the actions of phytoene, phytofluene or cyclolycopene cannot be ruled out since these compounds were present in most of the lycopene material used for these studies. Although lycopene remains as a promising agent, especially for cancer control, exploring interactions with other tomato phytochemicals and with current prostate cancer therapies should be encouraged.
Jin, Eun-Sun;Min, Joongkee;Jeon, Sang Ryong;Choi, Kyoung Hyo;Jeong, Je Hoon
Journal of Korean Neurosurgical Society
/
v.53
no.4
/
pp.207-212
/
2013
Objective : Recent studies have shown encouraging progress toward the use of autogenic and allogenic mesenchymal stem cells (MSCs) to arrest, or even lead to partial regeneration in, intervertebral disc (IVD) degeneration. However, this technology is still in its infancy, and further development is required. The aim of this study was to analyze whether rat adipose-derived mesenchymal stem cells (ADMSC) can differentiate towards IVD-like cells after treatment with transforming growth factor ${\beta}3$ (TGF-${\beta}3$) in vitro. We also performed quantitative analysis of gene expression for ADMSC only, ADMSCs treated with TGF-${\beta}3$, and co-cultured ADMSCs treated with TGF-${\beta}3$. Methods : ADMSCs were sub-cultured to homogeneity and used in fluorocytometry assays for CD11, CD45, and CD90/Thy1. ADMSCs were differentiated in spheroid culture towards the chondrogenic lineage by the presence of TGF-${\beta}3$, dexamethasone, and ascorbate. We also co-cultured pure ADMSCs and nucleus pulposus cells in 24-well plates, and performed immunohistochemical staining, western blotting, and RT-PCR for quantitative analysis of gene expression. Results : Results of fluorocytometry were positive for CD90/Thy1 and negative for CD11 and CD45. TGF-${\beta}3$-mediated induction of ADMSCs led to the expression of the differentiation markers of intervertebral disc-like cells, such as aggrecan, collagen II, and sox-9. Co-cultured ADMSCs treated with TGF-${\beta}3$ showed higher expression of differentiation markers and greater extracellular matrix production compared with ADMSCs treated with TGF-${\beta}3$ alone. Conclusion : ADMSC treated with TGF-${\beta}3$ may be an attractive source for regeneration therapy in degenerative IVD. These findings may also help elucidate the pathologic mechanism of MSC therapy in the degeneration of IVD in vivo.
Proceedings of the Korean Society of Embryo Transfer Conference
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2002.11a
/
pp.95-95
/
2002
Human embryonic stem (hES) cells can be induced to differentiate into tyrosine hydroxylase expressing (TH+) cells that may serve as an alternative for cell replacement therapy for Parkinson's disease (PD). To examine in vitro differentiation of hES (MB03, registered in NIH) cells into TH+ cells, hES cells were induced to differentiate according to the 4-/4+ protocol using retinoic acid (RA), ascorbic acid (AA), and/or lithium chloride (LiCl) followed by culture in N2 medium for 14 days, during which time the differentiation occurs. Immunocytochemical stainings of the cells revealed that approximately 21.1% of cells treated with RA plus AA expressed TH protein that is higher than the ratio of TH+ cells seen in any other treatment groups (RA, RA+LiCl or RA+AA+LiCl). In order to see the differentiation pattern in vivo and the ability of in vitro differentiation-induced cells in easing symptomatic motor function of PD animal model, cells (2 $\times$ 10$^{5}$ cells/2${mu}ell$) undergone 4-/4+ protocol using RA plus AA without any further treatment were transplanted into unilateral striatum of MPTP-lesioned PD animal model (C57BL/6). Following the surgery, motor behavior of the animals was examined by measuring the retention time on an accelerating rotar-rod far next 10 weeks. No significant differences in retention time of the animals were noticed until 2 weeks post-graft; however, it increased markedly at 6 weeks and 10 weeks time point after the surgery. Immunohistochemical studies confirmed that a reasonable number of TH+ cells were found at the graft site as well as other remote sites, showing the migrating nature of embryonic stem cells. These results suggest that in viかo differentiated hES cells relieve symptomatic motor behavior of PD animal model and should be considered as a promising alternative for the treatment of PD.
Background: The existing research results on the combined toxicity of these pollutants using mammals, such as rodents, are insufficient, especially in relation to changes in the immune system. Objectives: This study aims at evaluating the cellular immune response to PE-MPs solely or when combined with Pb, which possess excellent adsorption capacity with PE-MPs and is commonly co-exposed in our daily lives. Methods: The study investigated the cellular immune function of 9-week ICR mice with 28 days exposure to PE-MPs (2 mg/mouse/day) and Pb (0.1 mM in distilled water) individually and in combination. PE-MPs were administered via gastric intubation while the lead intake was conducted via the oral drinking water route. Cellular immunity was evaluated by analyzing the production for TH1 cytokines namely, TNF-α and IFN-𝛾 and TH2 cytokines, IL-4 and IL-6 in culture supernatants from polyclonally activated splenic mononuclear cells ex vivo. Results: Both the PE-MPs only and the PE-MPs+Pb exposure group revealed an increased TH1 response with elevated TNF-α and IFN-𝛾 levels and downregulated TH2 response with low IL-4, and IL-6 production levels compared to the control group. Furthermore, an increased IFN-𝛾/IL-4 ratio was found in the PE-MPs only and PE-MPs+Pb exposure groups, which indicated the skewedness to TH1 response. Meanwhile, reduced blood hemoglobin levels and increased levels of IL-4, the dominant TH2 cytokine in the Pb-only exposure group, were observed. Conclusions: Our current findings on the predominance of TH1 immune response in the PE-MPs and PE-MPs+Pb groups suggest that PE-MPs could be responsible for the predominant induction of the cellular immune changes. This finding could be used as an important landmark in research related to TH1 predominance, such as autoimmune diseases. It suggests that additional research on immune modulation using longer exposure durations or the same exposure route is required to elucidate stronger findings.
Objective: Hatching of the blastocyst from the zona pellucida (ZP) is a key event in mammalian implantation. In vivo, two factors have been identified as possible mediators of hatching: lysis of the ZP by substances elaborated either from the embryo or female reproductive tract and pressure exerted on the zona by expansion of the blastocyst. Two methods of zona manipulation were already in use to enhance the ability of embryos to hatch: mechanical PZD and chemical ZD by acidic Tyrode's solution. But several controversies of each method have been reported. The purpose of this study was to investigate the effect of pronase for mouse embryo hatching. Methods: Mouse embryos were obtained following ovulation induction of $F_1$ animals. Fresh and cryo-thawed morula embryos were exposed to 0.5, 1.0, 2.0, 5.0 ${\mu}g/ml$ pronase in Ham's F10 for 72 hrs. Main outcome measures were the rates of partial hatching and completely hatched blastocysts, and cell number of it. Results: In fresh and cryo-thawed group, the rates of completely hatched blastocyst were significantly higher in 5 ${\mu}g/ml$ pronase treatment group than control group. There was no difference in completely hatched blastocyst total cell number between pronase treatment group and control group. This suggest that pronase treatment did not harmful in mouse embryo development. In pronase treatment group, zona pellucida were thinner than control group. Conclusion: The addition of pronase to culture media may accelerate the hatching of embryo. So, enzymatic treatment of the zona may provide a valuable and effective assisted hatching technique for human in-vitro fertilization-embryo transfer.
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