• Title/Summary/Keyword: PC12h cells

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Molecular Characterization of Ischemia-Responsive Protein 94 (irp94) Response to Unfolded Protein Responses in the Neuron

  • Kim Seung-Whan;Kwon Ki-Sang;Shin Kee-Sun;Kim Seung-Ho;Kwon O-Yu
    • Biomedical Science Letters
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    • v.12 no.2
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    • pp.81-89
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    • 2006
  • The ischemia-responsive 94 gene (irp94) encoding a 94 kDa endoplasmic reticulum resident protein was investigated its molecular properties associated with unfoled protein responses. First, the expression of irp94 mRNA was tested after the reperfusion of the transient forebrain ischemia induction at the central nervous system in three Mongolian gerbils. Second, irp94 expression in PC12 cells, which are derived from transplantable rat pheochromocytoma cultured in the DMEM media, was tested at transcriptional and translational levels. The half life of irp94 mRNA was also determined In PC12 cells. Last, the changes of irp94 mRNA expression were investigated by the addition of various ER stress inducible chemicals (A23187, BFA, tunicamycin, DTT and $H_2O_2$) and proteasome inhibitors, and heat shock. High level expression of irp94 mRNA was detected after 3 hours reperfusion in the both sites of the cerebral cortex and hippocampus of the gerbil brain. The main regulation of irp94 mRNA expression in PC 12 cells was determined at the transcriptional level. The half life of irp94 mRNA in PC12 cells was approximately 5 hours after the initial translation. The remarkable expression of irp94 mRNA was detected by the treatment of tunicamycin, which blocks glycosylation of newly synthesized polypeptides, and $H_2O_2$, which induces apoptosis. When PC12 cells were treated with the cytosol proteasome inhibitors such as ALLN (N-acetyl-leucyl-norleucinal) and MG 132 (methylguanidine), irp94 mRNA expression was increased. These results indicate that expression of irp94 was induced by ER stress including oxidation condition and glycosylation blocking in proteins. Expression of irp94 was increased when the cells were chased after heat shock, suggesting that irp94 may be involved in recovery rather than protection against ER stresses. In addition, irp94 expression was remarkably increased when cytosol proteasomes were inhibited by ALLN and MG 132, suggesting that irp94 plays an important role for maintaining the ERAD (endoplasmic reticulum associated degradation) function.

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Protective Effect of PineXol® against Amyloid-β-induced Cell Death (아밀로이드 베타로 유도된 신경세포 사멸에 대한 PineXol®의 보호효과)

  • Han, Kyung-Hoon;Lee, Seung-Hee;Park, Kwang-Sung;Song, Kwan-Young;Kim, Jung-Hee;Park, Eun-Kuk;Han, Sung-Hee
    • The Korean Journal of Food And Nutrition
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    • v.30 no.6
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    • pp.1279-1285
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    • 2017
  • $Amyloid-{\beta}$ protein ($A{\beta}$) is known to increase free radical production in neuronal cells, leading to cell death by oxidative stress. The purpose of this study was to evaluate the protective effects of $PineXol^{(R)}$ on $A{\beta}_{25-35}$ induced neuronal cell death. Rat pheochromocytoma (PC-12) cells were pre-treated with $100{\mu}g/mL$ of $PineXol^{(R)}$ for 2 h. The cells were exposed to single dose of $30{\mu}M$ $A{\beta}_{25-35}$ for 24 h. Cell death was assessed by a cell count kit-8 (CCK-8) assay, lactate and dehydrogenase (LDH) release assay. An Apoptotic process was analyzed by a protein expression of the Bcl-2 family using western blotting. Cell viability increased in PC-12 cells treated with both $A{\beta}_{25-35}$ and $PineXol^{(R)}$, compared to the control group. $PineXol^{(R)}$ induced a decrease of the Bcl-2 protein expression (p<0.05), while Bax and Sod1 increased (p<0.05), indicating attenuation of $A{\beta}_{25-35}$ induced apoptosis. These results suggest that $PineXol^{(R)}$ may be a good candidate for the prevention of Alzheimer's disease(AD).

Non-toxic and Anti-oxydative effect of Dioscoreae Rhizoma on PC12 Cell (안태(安胎)에 활용되는 산약(山藥)의 신경세포주에 대한 안전성 및 항산화효과에 대한 연구)

  • Nam, Ju-Young;Roh, Jin-Ju;Seung, Jun-Ho;Son, Mi-Young;Khil, Mee-Jeong;Sung, Jung-Suk;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.4
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    • pp.61-76
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    • 2006
  • Purpose : This study examined the non-toxic and the anti-oxidative effect of Dioscoreae Rhizoma on PC12 cells. Sanyak(Dioscoreae Rhizoma; chinese yam, shan yao) is well-known for its curing power for kidney, lung, spleen. Tonifies and augments the spleen and stomach. Tonifies the lung gi and augments the kidney yin. Tonifies the kidneys and also stabilizes and binds. it also binds the essence and treats spermatorrhea, frequent urination, and vaginal discharge. We are therefore interested in whether Dioscoreae Rhizoma is capable of causing abnormal apoptosis processes, and whether this condition can be rectified through Dioscoreae Rhizoma herb treatment. Methods : We used aqueous extract to treat PC12 cells with different concentrations treated with a water or a MeOH extract of Dioscoreae Rhizoma (0, x10, x20, x40, x80). The MTT (3, (4, 5-dimethyl-thiazol) 2, 5-diphenyl-tetraxolium bromide) reduction assay was employed to quantify the differences in cell activity and viability. The Bax expression level was monitored using western-blotting techniques. The patterns of the changes in expression were scanned and analyzed. Results : Bax and GSK-3${\beta}$ promotes cell death and down-regulated during the development of the PC12 cells. This is indicated that Dioscoreae Rhizoma is capable of inducing apoptosis in PC12 cells. The induced cell death and significantly inhibited by Dioscoreae Rhizoma, which can be explained by the increase in the inhibition of Bax and GSK-3${\beta}$ expression. It was also shown that Dioscoreae Rhizoma inhibits the release of $H_2O_2$ and prevents lipid peroxidation. Furthermore, the accumulation of wild type Bax protein significantly downregulated in a dose-dependent manner upon treatment with Dioscoreae Rhizoma. Conclusion : In conclusion, Dioscoreae Rhizoma can induce apoptosis via a Bax-dependent pathway or GSK-3${\beta}$ dependent pathway in PC12 cells into anti-oxidant and protective effect.

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Hypothermia Regulates Insulin-like Growth Factor 1 Gene Expression in PC12 Cells

  • Yoo, Bo-Kyung;Kwon, Kisang;Lee, Eun Ryeong;Kim, Seung-Whan;Yu, Kweon;Kwon, O-Yu
    • Biomedical Science Letters
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    • v.23 no.1
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    • pp.39-43
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    • 2017
  • In this study, we evaluated the effects of various hypothermic conditions ($32^{\circ}C$), including lithium chloride treatment, on insulin-like growth factor 1 (IGF-1) gene expression in PC12 cells. The results show that short-term hypothermic treatment (<1 day) resulted in relatively higher IGF-1 gene expression than did longer-term treatment (>1 day). Repeated switching between normal temperature and hypothermia every 2 h increased IGF-1 gene expression approximately 3-4-fold. These findings indicate that hypothermia dynamically regulates IGF-1 gene expression. This study could be helpful for the development of treatment and diagnostic strategies for ischemia.

The Changes of Growth Patterns and the Production of Brain-Derived Neurotrophic Factors (BDNFs) in Perfusion Cultivation of Human Neuroblastoma Cells

  • Hong, Jong-Soo;Lee, Joo-Nho;Kim, Sun-Hee;Park, Kyung-Yoo;Cho, Jin-Sang;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.323-327
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    • 1999
  • It was shown that brain-derived neurotrophic factors (BDNFs) secreted from human neuroblastoma cells can significantly improve the growth of the neurites of PC12 nerve cells. The addition of purified BDNFs elongated the neurites of PC 12 nerve cells two to three times more than the case where the addition was not made. The perfusion rate strongly affected the change of the size of human neuroblastoma cells because the cell size decreased as the perfusion rate increased. This could also influence the productivity of BDNF from the cells. It is also important to note that the BDNF production was decreased when the cell size was reduced. BDNF production rate also decreased at a fast perfusion rate in a smaller cell size. At the relatively fast perfusion rate of 18 ml/h, the ratio of apoptotic to necrotic cells dramatically decreased, which possibly caused the decrease of BDNF production. It has been proven that the secretion of BDNF from human neuroblastoma cells was a partially growth-related process by yielding 6.2$\times l0^{-8}/g$ of BDNF/cell/h of growth related parameter and $0.48{\times}l0^{-9}/g$ of BDNF/cell/h of nongrowth-related parameter in a growth kinetic model. In addition, it was also found that the perfusion rate played a very important role in controlling the cell death mechanism.

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Protective Effect of Fucoidan Extract from Ecklonia cava on Hydrogen Peroxide-Induced Neurotoxicity

  • Park, Seon Kyeong;Kang, Jin Yong;Kim, Jong Min;Park, Sang Hyun;Kwon, Bong Seok;Kim, Gun-Hee;Heo, Ho Jin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.40-49
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    • 2018
  • We evaluated the antioxidant activity and neuronal cell-protective effect of fucoidan extract from Ecklonia cava (FEC) on hydrogen peroxide ($H_2O_2$)-induced cytotoxicity in PC-12 and MC-IXC cells to assess its protective effect against oxidative stress. Antioxidant activities were examined using the ABTS radical scavenging activity and malondialdehyde-inhibitory effect, and the results showed that FEC had significant antioxidant activity. Intracellular ROS contents and neuronal cell viability were investigated using the DCF-DA assay and MTT reduction assay. FEC also showed remarkable neuronal cell-protective effect compared with vitamin C as a positive control for both $H_2O_2$-treated PC-12 and MC-IXC cells. Based on the neuronal cell-protective effects, mitochondrial function was analyzed in PC-12 cells, and FEC significantly restored mitochondrial damage by increasing the mitochondrial membrane potential (${\Delta}{\Psi}m$) and ATP levels and regulating mitochondrial-mediated proteins (p-AMPK and BAX). Finally, the inhibitory effects against acetylcholinesterase (AChE), which is a critical hydrolyzing enzyme of the neurotransmitter acetylcholine in the cholinergic system, were investigated ($IC_{50}$ value = 1.3 mg/ml) and showed a mixed (competitive and noncompetitive) pattern of inhibition. Our findings suggest that FEC may be used as a potential material for alleviating oxidative stress-induced neuronal damage by regulating mitochondrial function and AChE inhibition.

Antioxidant Capacity and Protective Effects on Neuronal PC-12 Cells of Domestic Bred Kiwifruit (국내 육성 참다래의 항산화능 및 PC-12 신경세포 보호 효과)

  • Lee, Inil;Lee, Bong Han;Eom, Seok Hyun;Oh, Chang-Sik;Kang, Hee;Cho, Youn-Sup;Kim, Dae-Ok
    • Horticultural Science & Technology
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    • v.33 no.2
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    • pp.259-267
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    • 2015
  • This study was conducted to comparatively evaluate antioxidant capacity (AC) of seven cultivars of kiwifruit (Actinidia spp.) and their protective effects on neuronal PC-12 cells. The contents of total phenolics (TP) and total flavonoids (TF) of kiwifruits were also examined. Five cultivars of kiwifruit, Actinidia chinensis (cv. Haehyang and cv. Haegeum), A. eriantha (cv. Bidan), A. arguta ${\times}$ A. deliciosa (cv. Mansoo), and A. arguta (cv. Chiak), were bred in Korea, while two cultivars, A. deliciosa (cv. Hayward) and A. linguiensis (accession number 041AE), originated from New Zealand and China, respectively. Skin extracts of kiwifruit showed higher TP, TF, and AC than flesh extracts. The highest levels of TP and AC were found in cv. Bidan flesh extract among cultivars studied, but the TF content of cv. Bidan flesh extract was the lowest. The kiwifruit bred in Korea had higher AC than their counterparts. AC of kiwifruit had a highly positive linear correlation with TP and TF. The flesh extracts from cv. Hayward, cv. Haehyang, and cv. Haegeum significantly (p < 0.05) prevented PC-12 cells from oxidative stress induced using $H_2O_2$ compared to a control with $H_2O_2$ only. Overall, our results suggest that kiwifruit bred in Korea may offer a good source of antioxidants and serve as functional materials.

ACTIVATION OF P38 MAP KINASE AND AP-1 DURING THE PROMOTION OF NEURITE EXTENSION OF PC-12 CELLS BY 15-DEOXY-$\Delta$12,14-PROSTAGLANDIN J2

  • Song, Y.S.;Oh. J.H.;Park, K.S.;Jung, K.M.;Lee, M.K.;Jung, H.K.;Jung, S.Y.;Hong, J.T.
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.05a
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    • pp.110-110
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    • 2002
  • 15-Deoxy-$\Delta$12, 14-prostaglandin J2 (15-deoxy-PGJ2), a naturally occurring ligand activates the peroxisome proliferator-activated receptor-$\gamma$(PPAR-$\gamma$). It was known to have promoting ability of nerve growth factor(NGF)-induced neurite extension. However, it is not clear yet as to what signaling pathway is involved in its promoting ability of neurite extension.(omitted)

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Anti-tumor activities of Panax quinquefolius saponins and potential biomarkers in prostate cancer

  • He, Shan;Lyu, Fangqiao;Lou, Lixia;Liu, Lu;Li, Songlin;Jakowitsch, Johannes;Ma, Yan
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.273-286
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    • 2021
  • Background: Prostate carcinoma is the second most common cancer among men worldwide. Developing new therapeutic approaches and diagnostic biomarkers for prostate cancer (PC) is a significant need. The Chinese herbal medicine Panax quinquefolius saponins (PQS) have been reported to show anti-tumor effects. We hypothesized that PQS exhibits anti-cancer activity in human PC cells and we aimed to search for novel biomarkers allowing early diagnosis of PC. Methods: We used the human PC cell line DU145 and the prostate epithelial cell line PNT2 to perform cell viability assays, flow cytometric analysis of the cell cycle, and FACS-based apoptosis assays. Microarray-based gene expression analysis was used to display specific gene expression patterns and to search for novel biomarkers. Western blot and quantitative real-time PCR were performed to demonstrate the expression levels of multiple cancer-related genes. Results: Our data showed that PQS inhibited the viability of DU145 cells and induced cell cycle arrest at the G1 phase. A significant decrease in DU145 cell invasion and migration were observed after 24 h treatment by PQS. PQS up-regulated the expression levels of p21, p53, TMEM79, ACOXL, ETV5, and SPINT1 while it down-regulated the expression levels of bcl2, STAT3, FANCD2, DRD2, and TMPRSS2. Conclusion: PQS promoted cells apoptosis and inhibited the proliferation of DU145 cells, which suggests that PQS may be effective for treating PC. TMEM79 and ACOXL were expressed significantly higher in PNT2 than in DU145 cells and could be novel biomarker candidates for PC diagnosis.