• Journal of Life Science
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• v.21 no.2
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• pp.286-291
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• 2011

Hypoxia is a common condition found in a wide range of solid tumors and is often associated with metastasis and poor clinical outcomes. In the present study, we found that HIF-$1{\alpha}$ was induced by cobalt chloride (500 ${\mu}M$) treatment on human lung cancer cells, A549 and H460, for 24 hr. However, cobalt chloride (500 ${\mu}M$) did not affect cell proliferation of A549 and H460 in 48 hr. Cobalt chloride (500 ${\mu}M$) additionally induced epithelial-to-mesenchymal-like transition (EMT) such as reduced E-cadherin expression and increased ${\alpha}$-SMA expression. These results were confirmed by immunofluorecence experiment in H460 cells. E-cadherin was localized on the outer cell membrane. However, when the cells were treated with 500 ${\mu}M$ cobalt chloride for 24 hr, diffuse E-cadherin staining was observed, characteristic of a migratory mesenchymal phenotype. We also found that cobalt chloride induced integrin ${\beta}3$ expression and FAK phosphorylation in human lung cancer cells using western blotting and FACS anlaysis. Our data suggest that integrin ${\beta}3$-induced FAK phosphorylation may be developed into target molecules for blocking tumor metastasis.

• Journal of Dental Anesthesia and Pain Medicine
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• v.16 no.3
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• pp.175-184
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• 2016

Background: This study investigated the effect of remifentanil pretreatment on Cos-7 cells exposed to oxidative stress, and the influence of remifentanil on intracellular autophagy and apoptotic cell death. Methods: Cells were divided into 4 groups: (1) Control: non-pretreated cells were incubated in normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$). (2) $H_2O_2$: non-pretreated cells were exposed to $H_2O_2$ for 24 h. (3) RPC+$H_2O_2$: cells pretreated with remifentanil were exposed to $H_2O_2$ for 24 h. (4) 3-MA+RPC+$H_2O_2$: cells pretreated with 3-Methyladenine (3-MA) and remifentanil were exposed to $H_2O_2$ for 24 h. We determined the cell viability of each group using an MTT assay. Hoechst staining and FACS analysis of Cos-7 cells were performed to observe the effect of remifentanil on apoptosis. Autophagy activation was determined by fluorescence microscopy, MDC staining, and AO staining. The expression of autophagy-related proteins was observed using western blotting. Results: Remifentanil pretreatment increased the viability of Cos-7 cells exposed to oxidative stress. Hoechst staining and FACS analysis revealed that oxidative stress-dependent apoptosis was suppressed by the pretreatment. Additionally, fluorescence microscopy showed that remifentanil pretreatment led to autophagy-induction in Cos-7 cells, and the expression of autophagy-related proteins was increased in the RPC+$H_2O_2$ group. Conclusions: The study showed that remifentanil pretreatment stimulated autophagy and increased viability in an oxidative stress model of Cos-7 cells. Therefore, we suggest that apoptosis was activated upon oxidative stress, and remifentanil preconditioning increased the survival rate of the cells by activating autophagy.

• Polymer(Korea)
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• v.38 no.2
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• pp.156-163
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• 2014

Hydroxyapatite has osteoconductivity, biocompatibility and noninflammatory, and it has been used clinically as artificial bone. In this study, we prepared hydroxyapatite/poly(lactic-co-glycolic acid) (PLGA) scaffolds using 0, 10, 20, 40 and 60 wt% of hydroxyapatite. We analyzed compressive strength, SEM analysis and FTIR for mechanical property of 3D hydroxyapatite/PLGA scaffolds. For biocompatibility tests, cell proliferation and viability were measured via MTT assay and SEM. We analyzed RT-PCR, FACS, histology (H&E, ED-1) for anti-inflammatory effect. This study showed that hydroxyapatite hybrid scaffolds have low inflammatory reaction compared with the PLGA. This result has a potential for the application of artificial bone graft material.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.12 no.2
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• pp.20-52
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• 1999

In order to investigate the effect of Naetakchungumsankamibang(NTCGS) water extract on the skin tumor induced by 3-MCA and immunological responses in mice, the cytotoxicity against SK-MEL-2 cells and total number of tumors induced by 3-MCA were measured. The numbers of WBC, platelets and RBC, plaque forming cells, hemagglutinin titer, hemolysis titer, carbon clearance, proliferation of splenocyte by thymidine uptake assay, splenic leukocyte by FACS analysis and $TNF-{\alpha}$ were also measured for the evaluation of the immunological responses. The results were obtained as follows: 1. In cytotoxicity against SK-MEL-2 cells, concentration inhibiting cell growth up to below $20\%$ of control was recognized at 1mg/ml of NTCGS. 2. In Inhibitory effect on the skin tumor induced by 3-MCA, the results showed a strong inhibitory effect of NTCGS. 3. In hematological changes in the tumor bearing mice, the numbers of WBC decreased significantly in NTCGS treated group as compared with control. 4. In hematological changes in the tumor bearing mice, the numbers of platelets increased significantly in NTCGS treated group as compared with control. 5. In hematological changes in the tumor bearing mice, the numbers of RBC increased with no significance in NTCGS treated group as compared with control. 6. Effects of the plaque forming cells in the tumor bearing mice, NTCGS treated group exhibited a significant effect compared with control. 7. In terms of the effects on hemagglutinin titer, NTCGS treated group showed higher level than control, without significance. 8. In terms of the effects on hemolysis titer, NTCGS treated group showed higher level than control, without significance. 9. In terms of the effects on phagocytic index K in Balb/C mice, NTCGS treated group showed significant difference from control. 10. In terms of the effects on proliferation of splenocyte by thymidine uptake assay, NTCGS showed significant effect at the concentration of 0.5mg/ml. 11. In terms of the effects on splenic leukocyte of Balb/C mice by FACS analysis, NTCGS treated group showed significantly higher level of helper T cell, B cell and macrophage than in control. 12. In terms of the effects on the secretion of $TNF-{\alpha}$, the treated group showed significant effect at the concentration of 1mg/ml of NTCGS. Based on the results summarized above, NTCGS is considered to have antitumor activity and immunological responses against skin tumor, and to be usable fur the treatment.

• Journal of Life Science
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• v.28 no.11
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• pp.1316-1320
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• 2018

The effects of a green tea extract (GTE) were examined using the MCF-7 human breast cancer cell line. Cell viability assays using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays revealed that GTE had a significant cytotoxic effect on MCF-7 cells, depending on the concentration of GTE. Western blotting of p53 and its related proteins, p21/cip1 and CDK2, after GTE treatment revealed that a significant and concentration dependent increase in p53 protein in response to GTE. The levels of p21/cip1 proteins were also increased at low GTE concentrations were significantly increased even at the highest GTE concentrations. However, the level of CDK2 was significantly decreased by treatment with high concentrations of GTE. These results indicate that treatment with GTE increased the p53 level in MCF-7 cells, and this activation of p53 markedly elevated the levels of p21/cip1proteins, which, in turn, inhibited CDK2 expression in the MCF-7 cells. The inhibition of CDK2 expression might then affect cell cycle progression. Subsequent FACS analysis indicated that GTE treatment the gradually increased progression of the MCF-7 to the G1 phase. These results clearly demonstrate that the anti-tumor effect of GTE in MCF-7 cells is regulated by p53 arrest of the MCF-7 cells at the G1 stage of cell cycle.

• The Korea Journal of Herbology
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• v.33 no.4
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• pp.19-26
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• 2018

Objectives : The aim of this study was to investigate the immunopotentiating activity of combine extract that Silkworm and Cinnamomum cassia. Recently, acute epidemic diseases such as cold and viral respiratory diseases have been emerging. So, interested in immunity enhancement has been increasing, and research on natural products to promote immunity activity has been actively conducted. Methods : To confirm the immunopotentiating activity effect, Silkworm (SW), Cinnamomum cassia (CC), and SWCC combined extracts were treated 14 days at 300 mg/kg/day. The changes of glutamic oxalacetic transaminase (GOT), glutamic pyruvate transaminase (GPT) in serum were analyzed after experiment. The changes in the total spleen cell number were measured. Immune cells in spleen were analyzed using fluorescence activated cell sorter (FACS). also, analyzed the expression of cytokines in spleen. Results : Total number of cells in the spleen and FACS analysis of T lymphocytes activated in the spleen showed that the SWCC combined treated group had much higher frequency of active cells than both single groups. The ratio of CD4+CD8+, CD4+CD69+ and CD4+CD25+ T cells in spleen, SWCC is higher than other groups except Nor in CD4+, CD4+CD69+, CD4+CD25+ T cells. The results of this study suggest that SWCC can help immune function via IL-2, IL-10, IL-12, IFN-${\gamma}$ cytokine production, increased T lymphocytes and splenocyte proliferation. Conclusion : Therefore, these results suggested that the SWCC combined extracts administration increase stronger immunity enhancement than when SW and CC adminstration.

• Journal of Korean Academy of Nursing
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• v.38 no.2
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• pp.321-331
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• 2008

Purpose: To examine the relationship between body fat percentage (BFP) and N-K cell activity (NKCA) in Korean breast and rectal cancer patients just after diagnosis. Methods: With 35 subjects enrolled between November 2002 and May 2003, Bioelectrical Impedance Analysis was used to estimate BFP. FACS Analysis was used to measure N-K cell activity. The relationships between BFP and NKCA were identified by using curve estimation, simple regression, and multiple regression, Results: The mean BFPs of the subjects and all the sub-groups were higher than acceptable BFPs. Both the mean NKCAs of male and female subjects were lower than that of healthy women. NKCA was explained by BFP with a 14.9% variance in the total subjects (p<.05). There were significant negative relationships between BFP and NKCA after controlling age, type of cancer, and stage of cancer while no significant relationship was found after controlling for gender. The relationships between BFP and NKCA in the sub-groups of female, breast cancer, and stage I, and II were significant. The relationships between male, rectal cancer, and the stage III, and VI sub-groups were not identified, but they revealed a mild to moderate steep in curve estimation. Conclusion: Weight reduction could prevent the risk and advancement of breast and rectal cancer in Koreans.

• The Journal of Korean Medicine
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• v.25 no.4
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• pp.79-89
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• 2004

Saussurea lappa and Taraxacum mongolicum have been used for herbal medicinal treatments against cancers in East Asia. We performed this study to understand the molecular basis underlying the anti-tumor effects of two herbs and analyzed the effects of these medicinal herbs on proliferation and on expression of cell growth/apoptosis related molecules by using an AGS gastric cancer cell line. The treatments of Saussurea lappa dramatically reduced cell viabilities in a dose and time-dependent manner, but Taraxacum mongolicum did not. FACS analysis and Annexin V staining assay also showed that Saussurea lappa induces apoptotic cell death of AGS. Expression analyses via RT-PCR and Western blots revealed that Saussurea lappa increased expression of the p53 and its downstream effector p21/sup Waf1/, and that the both increased expression of apoptosis related Bax and cleavage of active caspase-3 protein. We also confirmed the translocation of Bax to mitochondria Collectively, our data demonstrate that Saussurea lappa induces growth inhibition and apoptosis of human gastric cancer cells, and these effects are correlated with down- and up-regulation of growth-regulating apoptotic and tumor suppressor genes, respectively.

• Korean Journal of Plant Resources
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• v.25 no.6
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• pp.693-699
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• 2012

This study was conducted to investigate the effects of Woohwangcheungsimweun (ox bezoar), deer antlers, and wild ginseng on induction of cardiomyocyte differentiation using the established mouse embryonic stem (ES) cells. The expression of atrial natriuretic peptide (ANP) was highest in Woohwangcheungsimweun treatment group. The expression of rabbit anti-GATA-4(GATA-4) and troponin (TnI) were highest in wild ginseng and Woohwangcheungsimweun treatment groups, respectively. Fluorescence activated cell sorting (FACS) analysis showed that the expression of ANP was highest in Dimethyl sulfoxide(DMSO) and Woohwangcheungsimweun treatment groups. The expression of GATA-4 was relatively high in wild ginseng treatment group. The expression of TnI was highest in Woohwangcheungsimweun treatment group. In the gene expression analysis, DMSO greatly inhibited GATA-4 expression to 25% of control. Woohwangcheungsimweun treatment caused to increase cTnI and cardiac ANP expression significantly. Wild ginseng extract upregulated GATA-4 gene expression. In conclusion, DMSO widely used as cardiomyocyte differentiation inducer did not show significant effects on the expression of ANP, GATA-4 and TnI in this study. Woohwangcheungsimweun showed upregulation of ANP and TnI expression. Wild ginseng extract showed greater effects than DMSO on GATA-4 expression. These results might suggest that the combination of Woohwangcheungsimweun and wild ginseng extract treatment can be expected to increase expressions of all three genes.

• IMMUNE NETWORK
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• v.9 no.6
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• pp.243-247
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• 2009

In this study, we report the development of a new dual reporter vector system for the analysis of promoter activity. This system employs green fluorescence emitting protein, EGFP, as a reporter, and uses red fluorescence emitting protein, DsRed, as a transfection control in a single vector. The expression of those two proteins can be readily detected via flow cytometry in a single analysis, with no need for any further manipulation after transfection. As this system allows for the simultaneous detection of both the control and reporter proteins in the same cells, only transfected cells which express the control protein, DsRed, can be subjected to promoter activity analysis, via the gating out of all un-transfected cells. This results in a dramatic increase in the promoter activity detection sensitivity. This novel reporter vector system should prove to be a simple and efficient method for the analysis of promoter activity.