• Title/Summary/Keyword: Fas and Fas-L

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β-elemene Induces Caspase-dependent Apoptosis in Human Glioma Cells in vitro through the Upregulation of Bax and Fas/FasL and Downregulation of Bcl-2

  • Li, Chen-Long;Chang, Liang;Guo, Lin;Zhao, Dan;Liu, Hui-Bin;Wang, Qiu-Shi;Zhang, Ping;Du, Wen-Zhong;Liu, Xing;Zhang, Hai-Tao;Liu, Yang;Zhang, Yao;Xie, Jing-Hong;Ming, Jian-Guang;Cui, Yu-Qiong;Sun, Ying;Zhang, Zhi-Ren;Jiang, Chuan-Lu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10407-10412
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    • 2015
  • Background: ${\beta}$-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of ${\beta}$-elemene against glioma cells remains unclear. In the present study, we assessed effects of ${\beta}$-elemene on human glioma cells and explored the underlying mechanism. Materials and Methods: Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of ${\beta}$-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of ${\beta}$-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and ${\beta}$-elemne treatment group. Results: With increase in the concentration of ${\beta}$-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability ($IC_{50}$) was $48.5{\mu}g/mL$ for 24h. ${\beta}$-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with ${\beta}$-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by ${\beta}$-elemene in a time and does-dependent manner. Conclusions: Our results indicate that ${\beta}$-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.

Effects of Red Ginseng Extract on Zearalenone Induced Spermatogenesis Impairment in Rat

  • Cho, Eun-Sang;Ryu, Si-Yun;Jung, Ju-Young;Park, Bae-Keun;Son, Hwa-Young
    • Journal of Ginseng Research
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    • v.35 no.3
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    • pp.294-300
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    • 2011
  • Zearalenone (ZEA) is a phenolic resorcylic acid lactone compound produced by several species of Fusarium. ZEA has toxic effects in the testes of domestic and laboratory animals. Korean red ginseng (KRG), the steamed root of Panax ginseng Meyer, has multiple pharmacological effects such as vasorelaxation, anti-thrombosis, anti-hypertension, etc. In this study, we investigated the effects of KRG extract on testicular toxicity induced by ZEA. Rats were treated with 300 mg/kg oral doses of KRG for 4 weeks every other day. The rats were then treated with a single dose of 5 mg/kg ZEA delivered intraperitoneally, whereas control rats received only doses of the vehicle. As a result, germ cell apoptosis induced by ZEA was decreased by KRG pre-treatment. In addition, Fas and Fas-L expression was reduced in rats that received KRG pre-treatment compared to ones treated with ZEA alone. In conclusion, impaired spermatogenesis resulting from ZEA treatment was prevented by KRG through Fas-Fas L modulating.

Platycodin D Induced NF-$textsc{k}$B Activation and Apoptosis in Immortalized Keratinocytes

  • Ahn, Kwang-Seok;Hahn, Bum-Soo;Lee, Eun-Bang;Kim, Yeong-Shik
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.195.3-196
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    • 2003
  • In this study, we investigated the molecular pathways targeted by platycodin D, which could involve apoptosis in immortalized human keratinocytes (HaCaT). We demonstrated that platycodin D-mediated apoptosis of HaCaT cells exhibited representative features, including DNA fragmentation, caspase-3, caspase-8 activation, and upregulation of Fas and FasL expression, but not p53 activation. To investigate the events involved in activation-induced FasL upregulation, we have examined mRNA accumulation, protein expression, and NF-$\kappa$B activity to elucidate transcription level in the HaCaT cell line treated with platycodin D. (omitted)

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Rhizoma Rehmanniae induced Apoptosis in Human Cervical Carcinoma HeLa Cells (생지황(生地黃)이 자궁경부암세포(子宮經部癌細胞)(HeLa cell)에 미치는 영향(影響))

  • Kim, Joo-Yeon;Jo, Ok-Hyon;Choe, Chang-Min;Cho, Han-Baek
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.1
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    • pp.69-80
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    • 2006
  • Purpose : This study is to evaluate the synergistic cytotoxicity of Rhizoma Rehmanniae(RR), in adriamycin-treated HeLa human cervical carcinoma cells. Methods : We culture HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% $CO_2$. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : The combination of RR and adriamycin synergistically augmented the cytotoxicity of HeLa cells. The apoptotic cell death was accompanied by the activation of caspase-3 and -8 as well as cleavage of poly(ADP- ribose) polymerase (PARP) in HeLa cells. The co-treatment of RR with adriamycin didn't have any effect on either the expression of Bcl-2 or that of Bax. Interestingly, a synergistic increase in apoptosis by the combination of two drugs was accompanied by the enhancement of Pas and Fas ligand (FasL) expression in HeLa cells. Taken together, the combination of RR and adriamycin significantly augmented the apoptotic cytotoxicity of Fas-positive cells, such as HeLa cells. The pathway is not involved in mitochondria-dependent pathway. Conclusion : RR induces apoptosis in HeLa cells via p38 MAPK activation.

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Effects of Compounds from Physalis angulata on Fatty Acid Synthesis and Glucose Metabolism in HepG2 Cells via the AMP-activated Protein Kinase Pathway

  • Hoa, Hoang Thai;Thu, Nguyen Thi;Dong, Nguyen Thuong;Oanh, Tran Thi;Hien, Tran Thi;Ha, Do Thi
    • Natural Product Sciences
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    • v.26 no.3
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    • pp.200-206
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    • 2020
  • The ability of the total extract from Physalis angulata; three fractions after partitioning with n-hexane, ethyl acetate (TBE), and water; and four withanolides (compounds 1 - 4) to phosphorylate 5'-adenosine monophosphate-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) in HepG2 cells was evaluated. The TBE fraction (50 ㎍/mL) activated p-ACC and p-AMPK expression most strongly. Compounds 1 - 4 (10 μM) upregulated p-ACC expression at different levels. Compound 4 induced the most significant changes in p-AMPK expression, followed by 1 and 2. Sterol regulatory element-binding proteins (SREBPs) play a functional role in the transcriptional regulation of the lipogenic pathway, including fatty acid synthase (FAS) and ACC. The effects of compounds 2 and 4 (10 μM) on FAS and SREBP-1c expression under high glucose conditions (30 mM) in HepG2 cells were evaluated further. Both dose-dependently inhibited FAS and SREBP-1c expression as well as lipid accumulation (1 - 10 μM) were compared to high-concentration glucose control, which upregulated FAS and SREBP-1c. These results suggest that compounds 2 and 4 upregulate AMPK, suppress FAS and SREBP-1c, and have potential effects on glucose and lipid metabolism.

Effects of Dietary Energy Density on Growth, Carcass Quality and mRNA Expression of Fatty Acid Synthase and Hormone-sensitive Lipase in Finishing Pigs

  • Liu, Z.H.;Yang, F.Y.;Kong, L.J.;Lai, C.H.;Piao, X.S.;Gu, Y.H.;Ou, X.Q.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.10
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    • pp.1587-1593
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    • 2007
  • A single factorial experiment was conducted to test the effects of three dietary levels of energy on mRNA expression of fatty acid synthase (FAS-mRNA) and hormone-sensitive lipase (HSL-mRNA) and their association with intramuscular fat in finishing pigs. 72 crossbred (Large $White{\times}Rongchang$) barrows with an average initial body weight of 20.71 (s.e. 0.1) kg, were randomly allotted to three dietary treatments (11.75, 13.05 and 14.36 MJ DE/kg) and fed until slaughtered at 100 or 101 kg. The diets were iso-nitrogenous and iso-essential amino acids. The growth performances including the duration of finishing were changed linearly (p<0.05) or quadratically (p<0.05) with increased dietary energy levels. The effects of dietary energy content on the percentage of external fat, intramuscular backfat and the fat thickness were linear (p<0.05). The content of dietary energy increased FAS-mRNA linearly or quadratically, while HSL-mRNA decreased linearly or quadratically in backfat and Longissmus dorsi muscle. Meanwhile, significant positive correlations (p<0.05) were found between energy level and intramuscular fat, FAS-mRNA or the ratio of FAS-mRNA to HSL-mRNA, between the ratio of FAS-mRNA to HSL-mRNA and intramuscular fat. However, the correlations between HSL mRNA and dietary energy or intramuscular fat were negative (p<0.05). The results indicated that dietary energy level regulates lipid accumulation, especially intramuscular fat, possibly by modulating the mRNA of FAS and HSL together rather than individually.

Trans Fatty Acid Isomers of Processed Foods Commonly Consumed in Korea (한국인 상용 가공식품의 trans 지방산 이성체)

  • 노경희;원미숙;송영선
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.3
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    • pp.325-337
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    • 2003
  • This study was designed to determine the distribution of trans fatty acids (tFAs) isomers of Processed foods commonly consumed in Korea. The tFAs positional isomers were analyzed using GC/MS spectrometer with HP-23 cis/trans FAME, capillary column (50m $\times$ 0.20 mm, id., 0.2 ${\mu}{\textrm}{m}$ film thickness) for 41 food samples. TFAs isomers were identified by comparing retention time with standards and GC/MS spectrum. In margarines, the content of tFAs ranged from 4.0% to 25.16% and the most abundant positional isomer of tFAs was C18:1 $\Delta$9t. In oils and fats, lards contained higher levels of tFAs (5.70~16.54%) than shortenings (6.77~10.55%). Shortenings contained higher levels of C18:1 $\Delta$9t (3.1~5.1%) than lard (1.6~4.3%), but corn oils had no tFAs. In seasonings, mayonnaise had no C16:1 $\Delta$9t, whereas C18:3t was detected. The content of tFAs in confectioneries was wide (16.20~52.16%). Among them, instant popcorns contained the highest amount of tFAs. Milk and dairy products showed even distribution of tFAS such as C18:1t, C18:2t, and C18:3t. Predominant tFAS isomer of condensed milk and ice cream was C16:1 $\Delta$9t. Frozen french fries and fried chicken contained higher levels of C18:1$\Delta$9t (9.4%), whereas grilled pork (jowl) had no C18:1 $\Delta$9t. The amount of tFAs per serving size was the highest in popcorn, followed by frozen pizza, frozen french fries, fried chicken, and bakeries.

Inducing Apoptosis of NCI-H157 Human Lung Carcinoma Cells via Activation of Caspase Cascade by Combination Treatment with Arsenic Trioxide and Sulindac (NCI-H157 폐암 세포주에서 Caspase Cascade 활성을 통한 Arsenic Trioxide와 Sulindac 병합요법의 세포고사효과)

  • Kim, Hak Ryul;Yang, Sei Hoon;Jeong, Eun Taik
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.4
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    • pp.381-392
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    • 2004
  • Arsenic trioxide($As_2O_3$) was introduced into the treatment of refractory or relapsed acute promyelocytic Ieukemia. Some investigators have reported that arsenic trioxide had induced apoptosis in a variety of solid human tumor cell lines, including non-small cell lung cancer. Non-steroidal anti-inflammatory drugs(NSAIDs) are powerful chemopreventive agents for gastrointestinal cancers and the growth of established tumors are reduced by inducing apoptosis. It's also reported that NSAIDs enhanced tumor response to chemotherapeutic drugs or radiation. In this study, we aimed to determine whether combination of arsenic trioxide with sulindac augmented its apoptotic potential in NCI-H157 human lung cancer cells. The human lung cancer cell line NCI-H157 was treated with arsenic trioxide and sulindac. Cell viability was measured by the MTT assay. Apoptosis was measured by nuclear staining and flow cytometric analysis. The catalytic activity of the caspase families were measured by the fluorogenic cleavage of biosubstrates. The western blotting were also performed to define the mechanical basis of apoptosis. Combination treatment of arsenic trioxide and sulindac decreased the viability of NCI-H157 human lung cancer cells in a dose-dependent manner. The catalytic activity of caspase-3, 8 and 9 proteases were increased after combination treatment. Consistently PARP was cleaved from 116kDa to 85kDa fragments, and the expression of ICAD was decreased by time-dependent manner. Also combination treatment increased the expression of Fas and Fas/L. Combination therapy of arsenic trioxide with sulindac augments cell death and induces apoptosis via the activation of caspase cascade in NCI-H157 human lung carcinoma cells.

Study on Transcriptional Factors Activation Change of HL-60 cell Apoptosis by Hedyotis Diffusa's Methanol Extract (백화사설초 메탄올 추출물에 의한 HL-60 세포고사과정에 있어서의 transcriptional factors 활성변화 연구)

  • Park, Sang-Goo;Lee, Ji-Hyun;Moon, Gu;Moon, Suk-Jae;Won, Jin-Hee;Park, Lae-Gil
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.6 no.1
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    • pp.67-79
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    • 2000
  • Objective : Hedyotis diffusa has been used as an anticancer agent for several decades in oriental medicine. We test whether the methanol extract of the herb affects transcriptional activation factors including $NF-{\kappa}B$ and AP-1. Methods : 1. HL-60 cells were treated with various concentrations(from 200 to $50{\mu}g/ml$) of methanol extract and $H_2O$ extract($200{\mu}g/ml$)of hedyotis diffusa, After 48h later, the cells were tested for viability by MTT assay. 2. The HL-60 cells were treated with $200{\mu}g/ml$ of methanol extract for the indicated periods. First. Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$ and AP-1. Second. Nuclear extracts were isolated and reacted with p50, p65. c-rel pan-Jun, c-Jun, JunB. JunD antibody on ice for 30min. Finally The cell lysates were prepared and analyzed by western blotting using anti-Fas, anti-FasL and anti-p53 antibody. Results : 1. The methanol extract decreases the viability of human lymphoid origin leukemia HL-60 cells in a dose-dependent manner. 2. $NF-{\kappa}B$ is rapidly activated by the addition of the methanol extract, reaches a peak at 30min and gradually returns to resting level. We confirm that $NF-{\kappa}B$ is a heterodimer mainly composed of p65 subunit with c-Rel. 3. Transcriptional activation of AP-1 is detected at 30min and reaches a maximum at 1hr after stimulation of the cells with the methanol extract. AP-1 is mainly composed with Jur-D and partially Jug-B proteins. 4. the methanol extract of Hedyotis diffusa induces the expression of Fas, Fas ligand and p53 proteins of HL-60 cells in a time dependent fashion. Conclusions : These results suggest that the methanol extract of Hedyotis diffusa exerts anticancer effects to induce the death of human leukomic HL-60 cells via activation of trascriptional factors such as $NF-{\kappa}B$ and AP-1, increase in expression of Fas mediated signalling proteins, and induction of tumor suppressor gene. p53.

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