• Korean Journal of Acupuncture
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• 제32권2호
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• pp.51-58
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• 2015

Objectives : Pinus densiflora gnarl, called Song-Jeol in Korean medicine, has been used to cure inflammatory diseases such as arthritis. In the present study, we evaluated inhibitory property of Song-Jeol pharmacopuncture(SJ) on C6 glioma cell migration. Methods : To evaluate cell viability on C6 glioma cells of SJ, the viability was assessed by using Ez-cytox assay kit. The cell migration was assessed by wound-healing assay and Boyden chamber assay, respectively. LPS-induced NO productions were determined by using the Griess reagent. The expression of iNOS and protein kinase $C(PKC)-{\alpha}$ were estimated by western blotting assay. Results : In the wound-healing assay and Boyden chamber assay, SJ showed a significant inhibition on serum-induced C6 glioma cell migration. In addition, NO production was decreased by SJ through suppression of iNOS expression in LPS-stimulated C6 glioma cell. Futhermore, LPS-induced protein kinase $C(PKC)-{\alpha}$ expression was effectively inhibited by SJ. Conclusions : These results demonstrated that SJ was useful for the suppression of the C6 glioma cell migration.

• Biomolecules & Therapeutics
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• 제8권2호
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• pp.140-146
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• 2000

Recently, we reported that immunostimulation of primary rat cortical astrocyte caused stimulation of glucose deprivation induced apoptotic cell death. To enhance the understanding of the mechanism of the potentiated cell death of clucose-deprived astrocyte by immunostimulation, we investigated the effect of immunostimulation on the glucose deprivation induced cell death of rat C6 glioma cells. Co-treatment of C6 glioma cells with lipopolysaccharide (LPS, $1\;{\mu}\textrm{g}/ml$) and interferon ${\gamma}(IFN{\gamma},\;100U/ml)$ is serum free condition caused marked elevationo f nitric oxide production ($>50\;{\mu}M$). In this condition, glucose deprivation caused significant release of lactate dehdrogenase (LDH) from C6 glioma cells while control cells did not show LDH release. To investigate whether elevated level of nitric oxide is responsible for the enhanced LDH release in glucose-deprived condition, C6 glioma cells were treated with 3-morphorinosydnonimine (SIN-1) and it was observed that SIN-1 caused increase in LDH release from glucose-deprived C6 glioma cells. Treatment of C6 glioma cells with $25\;{\mu}M$ of pyrrolidinedithiocarbamate (PDTC) which inhibit Nuclear factor kB (NF-kB) activation, caused complete inhibition of nitric oxide production. Treatment of C6 glioma cells with NO synthase inhibitors, $N^{G}$-nitro-L-arginine (NNA) or L-$N{\omega}$-nitro-L-arginine methyl ester (L-NAME), caused inhibition of nitric oxide production and also glucose deprivation induced cell death of cytokine-stimulated C6 glioma cells. In addition, diaminohydroxypyrimidine (DAHP, 5 mM) which inhibits the synthesis of tetrahydrobiopterine (BH4), one of essential cofactors for iNOS activity, caused complete inhibition of NO production from immunostimulated C6 glioma cells. The results from the present study suggest that immunostimulation causes potentiation of glucose deprivation induced death of C6 glioma cells which is mediated at least in part by the increased production of nitric oxide. The vulnerability of immunostimulated C6 glioma cells to hypoglycemic insults may implicate that the elevated level of cytokines in various ischemic and neurodegenerative diseases may play a role in their pathogenesis.

• Korean Journal of Acupuncture
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• 제30권4호
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• pp.212-219
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• 2013

Objectives : Cirsium japonicum is a traditional Korean medicine that has been used in the treatment of inflammatory diseases such as appendicitis, hepatitis, pulmonary abscess and tumor. The aim of study was to elucidate anti-migratory activity of CJP(Cirsium japonicum pharmacopuncture) through regulation of inflammatory mediators in C6 glioma cell. Methods : Nitric oxide(NO) production was determined by using nitrite assay. The cell migration was analyzed by wound-healing assay and Boyden chamber assay. The expression levels of iNOS, and protein kinase C(PKC)-${\alpha}$ were measured by western blotting assay. Results : CJP showed a significant decrease on NO production. Moreover, glioma cell migration was effectively suppressed by CJP. Furthermore, CJP inhibited the expressions of iNOS and PKC-${\alpha}$ in C6 glioma cells. Conclusions : These results suggest that CJP inhibits glioma cell migration and iNOS expression through regulation of PKC-${\alpha}$. Therefore, it is expected that CJP could be an effective agents for blocking malignant progression of glioma.

• 대한의생명과학회지
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• 제12권3호
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• pp.261-265
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• 2006

Toxic effect of Hexavalent chromium $(CrO_3)$ on various cells and organs has been well recognized. However, the mechanism and degree of cytotoxicity of $CrO_3$ remain unclear. This study was performed to examine the cytotoxicity of $CrO_3$ on $C_6$ glioma cells by measuring cell viability. The XTT assay, one of the sensitive methods to determine the cell viability, was taken to examine the viability of glioma cells treated with $CrO_3$. In this study, not only decreased the number of glioma cells but morphologic changes of them were noted and cell viability decreased in a time and dose-dependent manner after treated with various concentrations of $CrO_3$ for 48hours. $IC_{90}\;and\;IC_{50}$ values in XTT assay were determined at $25{\mu}M\;and\;55{\mu}M$ $CrO_3$, respectively. These results suggest that Hexavalent chromium has a highly cytotoxic effect and has a time and dose-dependent cytotoxicity on $C_6$ glioma cells.

• 대한의생명과학회지
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• 제12권4호
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• pp.451-455
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• 2006

It is demonstrated that inorganic mercury has cytotoxic effect on glial cells. Recently, oxygen radicals is involved in methylmercuric chloride (MMC)-induced cytotoxicity. But, the toxic mechanism of MMC is left unknown. The purpose of this study was to examine the cytotoxicity of MMC on $C_6$ glioma cells. The cytotoxicy was measured by cell viability using XTT assay in $C_6$ glioma cells. Colorimetric assay is regarded as a very sensitive screening method for the determination of the cell viability on various agents. In this study, MMC decreased cell viability according to the dose- and time dependent manners after $C_6$ glioma cells were grown with various concentrations of MMC for 48 hours. In the protective effect of allopurinol on MMC-induced cytotoxicity, allopurinol was effective in the prevention of MMC-induced cytotoxicity in these cultures. These results suggest that MMC has highly cytotoxic effect on $C_6$ glioma cells by the decrease of cell viavility, and free radical scavenger such as allopurinol was effective on organic mercury-induced cytotoxicity in these cultures.

• Nutrition Research and Practice
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• 제9권3호
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• pp.256-261
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• 2015

BACKGROUND/OBJECTIVES: Yacon (Samallanthus sonchifolius), a common edible plant grown throughout the world, is well known for its antidiabetic properties. It is also known to have several other pharmacological properties including anti-inflammatory, anti-oxidant, anti-allergic, and anti-cancer effects. To date, the effect of yacon on gliomas has not been studied. In this study, we investigated the effects of yacon on the migration and proliferation of C6 glioma cells stimulated by fetal bovine serum (FBS). MATERIALS/METHODS: Cell growth and proliferation were determined by evaluating cell viability using an EZ-Cytox Cell Viability Assay Kit. FBS-induced migration of C6 glioma cells was evaluated by performing the scratch wound healing assay and the Boyden chamber assay. We also used western blot analysis to determine the expression levels of extracellular signal-regulated kinase 1/2 (ERK1/2), a major regulator of migration and proliferation of glioma cells. Matrix metallopeptidase (MMP) 9 and TIMP-1 levels were measured by performing reverse transcription PCR. RESULTS: Yacon ($300{\mu}g/mL$) reduced both the FBS-induced proliferation of C6 glioma cells and the dose-dependent migration of the FBS-stimulated C6 cells. FBS-stimulated C6 glioma cells treated with yacon (200 and $300{\mu}g/mL$) showed reduced phosphorylation of ERK1/2 and inhibition of MMP 9 expression compared to those shown by the untreated FBS-stimulated C6 cells. In contrast, yacon (200 and $300{\mu}g/mL$) induced TIMP-1 expression. CONCLUSIONS: On the basis of these results, we suggest that yacon may exert an anti-cancer effect on FBS-stimulated C6 glioma cells by inhibiting their proliferation and migration. The most likely mechanism for this is down-regulation of ERK1/2 and MMP9 and up-regulation of TIMP-1 expression levels.

• 동의생리병리학회지
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• 제28권4호
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• pp.396-402
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• 2014

This study is designed to investigate cytoprotective effects of Platycodon grandiflorus (Jacq.) A.DC on C6 glioma cell apoptosis by oxidative stress. Experimental C6 glioma cells were classified into four groups as follows: normal group, PGE group, chemical groups, PGE+chemical groups. Oxidative stress that caused by chemicals in the C6 glioma cell, check the impact to Chemical group was administered normal group. Apoptotic effect protecting in order to observe the chemical group was administered PGE. We to observe effects of PGE on SOD inhibition, total glutathione production in C6 glioma cells were administered PGE. In case of administration PGE, apoptosis induced by Paraquat was significantly decreased. In case of administration PGE, apoptosis induced by SNP was significantly decreased. In case of administration PGE, apoptosis induced by $H_2O_2$ was significantly decreased. In case of administration PGE, apoptosis induced by Rotenone was decreased, but the statistical significance was not. In case of administration PGE, SOD inhibition activities significantly decreased. In case of administration PGE, Total glutathione did not affect the content. These results suggest that PGE is able to treat a disease caused by oxidative stress and prevent a aging. These results suggest that PGE is a disease caused by oxidative stress and aging, the prevention and treatment of food shall be able to be applied.

• The Korean Journal of Physiology and Pharmacology
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• 제2권4호
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• pp.419-425
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• 1998

To clarify the effect of extracellular ATP in cultured C6 glioma cells, ATP-induced cytosolic free calcium ($[Ca^{2+}]_i$) mobilization and cell proliferation were investigated. ATP-induced $[Ca^{2+}]_i$ increased in a dose-dependent manner $(10^{-7}\;M{\sim}10^{-3}\;M)$. ATP-induced $[Ca^{2+}]_i$ increases were slightly slowed in extracellular calcium-free conditions especially in sustained phase. ATP-induced $[Ca^{2+}]_i$ increment was also inhibited by the pretreatment of U73122, a phospholipase C (PLC) inhibitor, in a time-dependent manner. Suramin, a putative $P_{2Y}$ receptor antagonist, dose-dependently weakened ATP-induced $[Ca^{2+}]_i$ mobilization. Significant increases in cell proliferation were observed at 2, 3, and 4 days after ATP was added. Stimulated cell proliferation was also observed with adenosine at days 2 and 3. This cell proliferation was significantly inhibited by the treatment with suramin. Ionomycin also stimulated cell proliferation in a concentration-dependent manner. In conclusion, we suggest that extracellular ATP stimulates C6 glioma cell proliferation via intracellular free calcium mobilization mediated by purinoceptor.

• 대한의생명과학회지
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• 제12권3호
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• pp.275-279
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• 2006

It is demonstrated that cadmium has cytotoxic effect on glial cells, oxygen radicals are involved in cadmium-induced cytotoxicity. However, the toxic mechanism of cadmium is left unknown so far. The purpose of this study was to examine the cytotoxicity of $CdCl_2$ on $C_6$ glioma cells. The cytotoxicy was measured by cell viability via XTT assay in $C_6$ glioma cells. Colorimetric assay such as XTT assay is regarded as a very sensitive screening method for the determination of the cell viability on a lots of chemicals. In this study, $CdCl_2$ decreased cell viability according to the dose- and time dependent manners after $C_6$ glioma cells were treated with various concentrations of $CdCl_2$ for 48 hours. $IC_{90}\;and\;IC_{50}$ values for XTT assay was determined at $5{\mu}M\;and\;55{\mu}M$ of $CdCl_2$, respectively. These results suggest that $CdCl_2$ has highly cytotoxic effect on $C_6$glioma cells by the decrease of cell viability.

• Nutrition Research and Practice
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• 제10권3호
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• pp.259-264
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• 2016

BACKGROUND/OBJECTIVES: Stromal cell-derived growth factor 1 (SDF-1), also known as chemokine ligand 12, and chemokine receptor type 4 are involved in cancer cell migration. Compound K (CK), a metabolite of protopanaxadiol-type ginsenoside by gut microbiota, is reported to have therapeutic potential in cancer therapy. However, the inhibitory effect of CK on SDF-1 pathway-induced migration of glioma has not yet been established. MATERIALS/METHODS: Cytotoxicity of CK in C6 glioma cells was determined using an EZ-Cytox cell viability assay kit. Cell migration was tested using the wound healing and Boyden chamber assay. Phosphorylation levels of protein kinase C $(PKC){\alpha}$ and extracellular signal-regulated kinase (ERK) were measured by western blot assay, and matrix metallopeptidases (MMP) were measured by gelatin-zymography analysis. RESULTS: CK significantly reduced the phosphorylation of $PKC{\alpha}$ and ERK1/2, expression of MMP9 and MMP2, and inhibited the migration of C6 glioma cells under SDF-1-stimulated conditions. CONCLUSIONS: CK is a cell migration inhibitor that inhibits C6 glioma cell migration by regulating its downstream signaling molecules including $PKC{\alpha}$, ERK1/2, and MMPs.