• Title/Summary/Keyword: C6 cell

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Cellular Mechanism of Newly Synthesized Indoledione Derivative-induced Immunological Death of Tumor Cell

  • Oh, Su-Jin;Ryu, Chung-Kyu;Baek, So-Young;Lee, Hyun-Ah
    • IMMUNE NETWORK
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    • v.11 no.6
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    • pp.383-389
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    • 2011
  • Background: EY-6 is one of the newly synthesized indoledione derivatives to induce tumor cell-specific cell death. In this study, we investigated the mechanism of immunological death induced by EY-6 at mouse colon cancer cell as well as at the normal immune cell represented by dendritic cell. Methods: C57BL/6 mouse syngeneic colon cancer cell MC38 was treated with EY-6, and analyzed by MTT for viability test, flow cytometry for confirming surface expressing molecules and ELISA for detection of cytokine secretion. Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC. Results: EY-6 killed the MC38 tumor cells in a dose dependent manner (25, 50 and $100{\mu}M$) with carleticulin induction. And EY-6 induced the secretion of IFN-${\gamma}$ but not of TNF-${\alpha}$ from the MC38 tumor cells. EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation. The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation). Conclusion: Data indicate that the EY-6 induced tumor cell specific and immunological cell death by modulation of tumor cell phenotype and cytokine secretion favoring induction of specific immunity eliminating tumor cells.

Anti-proliferating Effects of Porphyra tenera Fractions on Several Cancer Cell Lines in uitro (김 분획물의 in vitro에서의 항발암효과)

  • Shin, Mi-Ok;Bae, Song-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.10
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    • pp.1514-1519
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    • 2005
  • This study was performed to investigate the effects of Porphyra tenera (PT) on cytotoxicity and quinone reductase (QR) activity in the cancer cells. PT was extracted with methanol and further fractionated into five different types: hexane (PTMH), ethyl-ether (PTMEE), ethylacetate (PTMEA) butanol (PTMB) and aquous (PTMA) partition layers. We determined the cytotoxic effect of these layers on C6, HepG2, MCF-7, and HT-29 cell lines by MTT assay. Among the various fractions, hexane (PTMH) of PT showed the strongest cytotoxic effect on C6, HepG2 and MCF-7 cell lines. PTMH displayed very low level of cytotoxicity at the lower concentration levels and at 300 $\mu$g/mL. PTMH resulted in 87.5$\%$ growth inhibition on C6 cell 70 $\%$ on the HepG2 cell and 89$\%$ on the MCF-7 cell, which were significantly high compared to other fractions. A 400 $\mu$g/mL PTMH concentration level, 99$\%$, 94.5$\%$ and 99$\%$ of cell growth inhibition were resulted on the same cell lines. On HT-29 cell line, both hexane (PTMH) and aqueous (PTMA) fraction of PT showed cytotoxic effects, but the Percentage was not as high as previous results tested on other cell lines such as C6 HepG2 and MCF-7 cell lines. Also, we observed quinone reductase (QR) inducing-effects in all fractions of PT on HepG2 cells. The QR inducing effects of the PTMH on HepG2 cells at 150 $\mu$g/mL concentration was 6.6 times higher than the control. Although further studies are needed, the present work suggests that PT was a potential to be used as a chemopreventive.

Effect of Cadmium on $C_6$ Glioma Cells in Culture

  • Son Young-Woo;Lee Kang-Chang
    • Biomedical Science Letters
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    • v.12 no.3
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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.

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Protective Effects of Ukgan-san in $CoCl_2$-induced Cell Death of C6 Glial Cells ($CoCl_2$로 유도된 C6 신경교세포의 사멸에 대한 억간산(抑肝散)의 보호 효과)

  • Cho, Mun-Young;Shin, Yong-Jeen;Ha, Ye-Jin;Woo, Chan;Kim, Ta-Jung;You, Ju-Yeon;Choi, Yong-Seok;Choi, Jung-Hoon;Shin, Sun-Ho
    • The Journal of Internal Korean Medicine
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    • v.34 no.2
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    • pp.178-191
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    • 2013
  • Objectives : In this study, we made an effort to investigate the protective mechanism of Ukgan-san (UGS) extracts on hypoxia-induced C6 glial cell death. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were analysed with microscope after staining with crystal violet (CV). Reactive oxygen species (ROS) formation was assessed by flow cytometer after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). We also analyzed expression of hypoxia-inducible factor-1 alpha (HIF-$1{\alpha}$) and p53, processing of procaspase-3 and procyclic acidic repetitive protein (PARP) by western blot method. Results : We estimated the elevated cell viability by UGS extract on $CoCl_2$-induced C6 glial cells. UGS attenuated $CoCl_2$-induced ROS formation in C6 glial cells and also showed a protective activity compared to antioxidants and exhibited abrogation of LDH-released by $CoCl_2$. UGS suppressed the typical apoptotic cell death markers, caspase-3 and PARP activation. UGS inhibited $CoCl_2$-induced HIF-1${\alpha}$ expression which is known as a major regulator for hypoxia-induced cell death, and suppressed p53 expression. Conclusions : These results suggest that UGS extract contains protective constituents for hypoxia-induced C6 glial cell death.

Identification of the Marker-Genes for Dioxin(2, 3, 7, 8- tetradibenzo-p-dioxin)-Induced Immune Dysfunction by Using the High-Density Oligonucleotide Microarray

  • Kim, Jeong-Ah;Lee, Eun-Ju;Chung, In Hye;Kim, Hyung-Lae
    • Genomics & Informatics
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    • v.2 no.2
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    • pp.75-80
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    • 2004
  • In a variety of animal species, the perinatal exposure of experimental animals to the 2,3,7,8-tetrachlorodibenzo­p-dioxin (TCDD) leads to the immune dysfunction, which is more severe and persistent than that caused by adult exposure. We report here the changes of gene expression and the identification of the marker-genes representing the dioxin exposure. The expressions of the transcripts were analyzed using the 11 K oligonucleotide­microarray from the bone marrow cells of male C57BL/6J mice after an intraperitoneal injection of $1{\mu}g$ TCDD/kg body weight at various time intervals: gestational 6.5 day(G6.5), 13.5 day(G13.5), 18.5 day(G18.5), and postnatal 3 (P3W)and 6 week (P6W). The type of self-organizing maps(SOM) representing the specific exposure dioxin could be identified as follows; G6.5D(C14), G13.5D(C0, C5, C10, C18), G18.5D(7): P3W(C2, C21), and P6W(C4, C15, C20). The candidate marker-genes were restricted to the transcripts, which could be consistently expressed greater than $\pm$2-fold in three experiments. The resulting candidates were 85 genes, the characteristics of that were involved in cell physiology and cell functions such as cell proliferation and immune function. We identified the biomarker-genes for dioxin exposure: smc -like 2 from SOM C14 for the dioxin exposure at G6.5D, focal adhesion kinase and 6 other genes from C0, and protein tyrosine phosphatase 4a2 and 3 other genes from C5 for G13.5D, platelet factor 4 from C7 for G18.5D, fos from C2 for P3W.

SCP Production from Mandarin Orange Peel Press Liquor (감귤과피 압착액을 기질로 한 SCP 생산)

  • 강신권;성낙계
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.556-562
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    • 1989
  • The bioconversion of mandarin orange peel press liquor to single cell protein (SCP) by two yeast strains, F-60, and C-7, which were isolated from mandarin orange peel was carried out and compared with that of using Candida utilis IFO 0598. Experiments were directed toward the high yield of biomass and high protein in cultures of the strains mentioned above. Candida utilis IFO 0598, F-60 and C-7 strains were cultivated at 3$0^{\circ}C$, pH 5.2 for 3 days in shaking flasks. The effects of some nutrients on cell growth were studied. Cell mass and protein content per cell mass were increased by addition of urea 1%, KH$_2$PO$_4$ 0.1% and MgSO$_4$ㆍ7$H_2O$ 0.05%, When the F-60 strain cultured under the optimal conditions, cell mass, growth yield and protein content were 41.2g/l, 53.9%, 59.7%, respectively. Cell mass was also increased up to 15% by modifying the fermentation condition on the bench type 20l jar fermentor. Crude fat content (10.3%) of dried C-7 cell was higher than those of C. utilis and F-60, 4.9% and 5.6% respectively. Total protein content of the F-60 strain was 59.7% per dry weight. And we compared their amino acid compositions with that of FAO provisional pattern. In the case of the F-60 strains, amino acid contents such as lysine, leucine and isoleucine were much higher than those of methionine, cystine and tryptophan.

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Response of Metastatic Cancer Cells to Thermal Changes in vitro (배양온도 변화에 대한 전이성 암세포의 반응)

  • Ahn, San-Gil;Kwon, Young-Ee;Choi, Ho-Soon;Kwon, Jung-Kyun;Yoo, Jin-Young;Kim, Jong-Ryong;Kim, Won-Kyu
    • Applied Microscopy
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    • v.37 no.4
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    • pp.239-248
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    • 2007
  • Alteration of temperature is one of cancer therapies. In general, severe hyperthermia(around $43^{\circ}C$) and hypothermia(around $18^{\circ}C$) trigger apoptosis through mitochondria, though the specific mechanism is still unknown. CC-t6 and GB-d1 cell lines, which were originally derived from human cholangiocarcinoma and gall bladder cancer, were established from a metastatic lymph node. To investigate the mechanism of metastatic cancer cell response to thermal stresses, hyperthermia($37^{\circ}C{\rightarrow}43^{\circ}C$) and hypothermia($37^{\circ}C{\rightarrow}17.4^{\circ}C$) were designed. Thermal stresses did not induce apoptosis but necrotic cell death. Any alterations of caspase-3, -9, cytochrome c, Bax, and Bcl-2 were not found in both hyperthermia and hypothermia exposed fells using western blot analysis. In the transmission electron microscopy, typical necrotic, but not apoptotic, changes were observed. These results suggest that temperature changes induce cell death through necrotic pathway in metastatic cancer in vitro, and it can be one of effective anticancer methods.

Aucklandia lappa Causes Cell Wall Damage in Candida albicans by Reducing Chitin and (1,3)-β-D-Glucan

  • Lee, Heung-Shick;Kim, Younhee
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.967-973
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    • 2020
  • The fungal cell wall is a major target of antifungals. In this study, we report the antifungal activity of an ethanol extract from Aucklandia lappa against Candida albicans. We found that the extract caused cell wall injury by decreasing chitin synthesis or assembly and (1,3)-β-D-glucan synthesis. A sorbitol protection assay demonstrated that the minimum inhibitory concentration (MIC) of the A. lappa extract against C. albicans cells increased eight-fold from 0.78 to 6.24 mg/ml in 72 h. Cell aggregates, which indicate damage to the cell wall or membrane, were commonly observed in the A. lappatreated C. albicans cells through microscopic analysis. In addition, the relative fluorescence intensities of the C. albicans cells incubated with the A. lappa extract for 3, 5, and 6 h were 92.1, 84.6, and 79.8%, respectively, compared to the controls, estimated by Calcofluor White binding assay. This result indicates that chitin content was reduced by the A. lappa treatment. Furthermore, synthesis of (1,3)-β-D-glucan polymers was inhibited to 84.3, 79.7, and 70.2% of that of the control treatment following incubation of C. albicans microsomes with the A. lappa extract at a final concentration equal to its MIC, 2× MIC, and 4× MIC, respectively. These findings suggest that the A. lappa ethanol extract may aid the development of a new antifungal to successfully control Candidaassociated disease.

Protective Effect of Korean Red Ginseng against 6-Hydroxydopamine-induced Nitrosative Cell Death via Fortifying Cellular Defense System (6-Hydroxydopamine으로 유도된 질소적 세포 사멸에 대한 고려홍삼 추출물의 보호효과)

  • Lee, Chan;Jang, Jung-Hee;Park, Gyu Hwan
    • YAKHAK HOEJI
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    • v.60 no.2
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    • pp.92-99
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    • 2016
  • Parkinson's disease (PD) is one of the representative neurodegenerative movement disorders with the selective loss of dopaminergic neurons in the substantia nigra. 6-Hydroxydopamine (6-OHDA) is widely used as an experimental model system to mimic PD and has been reported to cause neuronal cell death via oxidative and/or nitrosative stress. Therefore, daily intake of dietary or medicinal plants which fortifies cellular antioxidant capacity can exert neuroprotective effects in PD. In the present study, we have investigated the protective effect of Korean red ginseng (KRG) against 6-OHDA-induced nitrosative death in C6 glioma cells. Treatment of C6 cells with 6-OHDA decreased cell viability and increased expression of inducible nitric oxide synthase, production of nitric oxide as well as peroxynitrite, and formation of nitrotyrosine. 6-OHDA led to apoptotic cell death as determined by decreased Bcl-2/Bax, phosphorylation of JNK, activation of caspase-3, and cleavage of PARP. Conversely, pretreatment of C6 cells with KRG attenuated 6-ODHA-induced cytotoxicity, apoptosis, and nitrosative damages. To further elucidate the molecular mechanism of KRG protection against 6-OHDA-induced nitrosative cell death, we have focused on the cellular self-defense molecules against exogenous noxious stimuli. KRG treatment up-regulated heme oxygenase-1 (HO-1), a key antioxidant enzyme essential for cellular defense against oxidative and/or nitrosative stress via activation of Nrf2. Taken together, these findings suggest KRG may have preventive and/or therapeutic potentials for the management of PD.

The Effect of Korean Ginseng on the Immunotoxicity of Mitomycin C (Mitomycin C의 면역독성에 미치는 고려인삼의 영향)

  • 안영근;김주영;정종갑;김정훈;구자돈
    • YAKHAK HOEJI
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    • v.31 no.6
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    • pp.355-360
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    • 1987
  • The immunopotenciating effects of ethanol extract, butanol fraction and petroleum ether extract of Korean Ginseng on the immunotoxicity of mitomycin C were investigated in mice. A single administration of mitomycin C induced an apparent but relatively transient reduction of spleen weight, hemagglutination titer, Arthus reaction, RFC and natural killer cell activity. Ethanol extract of Ginseng significantly restored spleen weight, HA titer, RFC and natural killer cell activity. Butanol fraction of Ginseng significantly restored HA titer, RFC and natural killer cell activity. Petroleum ether extract of Ginseng very significantly restored spleen weight, Arthus reaction, DTH, RFC and natural killer cell activity.

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