• Title/Summary/Keyword: 14-3-3 isoform

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Dexamethasone Downregulates Expressions of 14-3-3β and γ-Isoforms in Mice with Eosinophilic Meningitis Caused by Angiostrongylus cantonensis Infection

  • Tsai, Hung-Chin;Chen, Yu-Hsin;Yen, Chuan-Min;Chung, Li-Yu;Wann, Shue-Ren;Lee, Susan Shin-Jung;Chen, Yao-Shen
    • Parasites, Hosts and Diseases
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    • v.57 no.3
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    • pp.249-256
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    • 2019
  • Steroids are commonly used in patients with eosinophilic meningitis caused by A. cantonensis infections. The mechanism steroids act on eosinophilic meningitis remains unclear. In this mouse experiments, expressions of 14-3-3 isoform ${\beta}$ and ${\gamma}$ proteins significantly increased in the CSF 2-3 weeks after the infection, but not increasedin the dexamethasone-treated group. Expression of 14-3-3 ${\beta}$, ${\gamma}$, ${\varepsilon}$, and ${\theta}$ isoforms increased in brain meninges over the 3-week period after infection and decreased due to dexamethasone treatment. In conclusion, administration of dexamethasone in mice with eosinophilic meningitis decreased expressions of 14-3-3 isoform proteins in the CSF and in brain meninges.

14-3-3-Associated Proteins in Helicobacter pylori-Infected Gastric Epithelial Cells (Helicobacterpylori에 감염된 위상피세포에서 14-3-3 결합 단백질의 변화)

  • Chung, Hae-Yun
    • The Korean Journal of Food And Nutrition
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    • v.24 no.2
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    • pp.258-267
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    • 2011
  • 14-3-3 is a highly conserved, ubiquitously expressed protein family. It associates with diverse cellular proteins through its specific phosphoserine/phosphothreonine-binding activity and thus contributes to the regulation of crucial cellular processes such as metabolism, signal transduction, cell-cycle control, apoptosis, protein trafficking, transcription and stress responses. This study aims to determine changes in levels of 14-3-3 isoforms and 14-3-3 - associated proteins in Helicobacter pylori(H. pylori)-infected gastric epithelial AGS cells. AGS cells were stimulated with H. pylori(NCTC 11637) at the ratio of 300:1(bacterium:cell). Western blot analysis revealed that 14-3-3 $\sigma$ was elevated at 3 hr after H. pylori treatment. Other isoforms were not significantly affected by H. pylori infection. Using immunoprecipitation to 14-3-3 $\sigma$, followed by proteomic analysis, we found that S phase kinase associated protein isoform 2 bound to 14-3-3 $\sigma$ has increased. In contrast, three proteins (DEAD-box polypeptide 3, heterogeneous nuclear ribonucleoprotein H2 and WD repeat-containing protein isoform 1) bound to 14-3-3 decreased by H. pylori infection. Our results suggest that 14-3-3 may play an important regulatory role in H. pylori-induced signal transduction in gastric epithelial cells.

The Clinicopathological and Prognostic Impact of 14-3-3 Protein Isoforms Expression in Human Cholangiocarcinoma by Immunohistochemistry

  • Wu, Qiao;Liu, Chang-Zheng;Tao, Lian-Yuan;Yu, Lan;Liu, Wei;Chen, Song-Sen;He, Xiao-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1253-1259
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    • 2012
  • The 14-3-3 proteins are highly conserved, ubiquitous molecules involved in a variety of biologic phenomena, such as cell cycle control, and apoptosis. However, their expression in cholangiocarcinoma has not been previously characterized. In this paper, immunohistochemistry using specific anti-14-3-3 monoclonal antibodies was performed on formalin-fixed;, paraffin embedded archival tissue from 86 patients of cholangiocarcinoma. We also examined the correlation between expression and survival rate and clinicopathologic factors such as tumor location, tumor size, pathologic differentiation, lymphatic permeation, lymph node metastasis, and tumor stage. Positive 14-3-3 proteins expression was observed for 6 isoforms (${\beta}$, ${\sigma}$, ${\gamma}$, ${\theta}$, ${\delta}$, ${\eta}$) of these proteins in 86 patients of cholangiocarcinoma. ${\beta}$ and ${\sigma}$ isoform immunoreactivity was correlated with lymph node metastasis, tumor stage and patients' survival rate. In addition, ${\delta}$ isoform immunoreactivity showed trends with tumor location, tumor size, pathologic differentiation and tumor stage, while the ${\theta}$ isoform was correlated with pathologic differentiation. These results indicated that upregulated expression of some isoforms of 14-3-3 may be a common mechanism for evading apoptosis in cholangiocarcinoma, so that targeting 14-3-3 may be a novel promising strategy for the treatment of this tumor.

R-type Calcium Channel Isoform in Rat Dorsal Root Ganglion Neurons

  • Fang, Zhi;Hwang, Jae-Hong;Kim, Joong-Soo;Jung, Sung-Jun;Oh, Seog-Bae
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.1
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    • pp.45-49
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    • 2010
  • R-type $Ca_v2.3$ high voltage-activated $Ca^{2+}$ channels in peripheral sensory neurons contribute to pain transmission. Recently we have demonstrated that, among the six $Ca_v2.3$ isoforms ($Ca_v2.3a{\sim}Ca_v2.3e$), the $Ca_v2.3e$ isoform is primarily expressed in trigeminal ganglion (TG) nociceptive neurons. In the present study, we further investigated expression patterns of $Ca_v2.3$ isoforms in the dorsal root ganglion (DRG) neurons. As in TG neurons, whole tissue RT-PCR analyses revealed the presence of two isoforms, $Ca_v2.3a$ and $Ca_v2.3e$, in DRG neurons. Single-cell RT-PCR detected the expression of $Ca_v2.3e$ mRNA in 20% (n=14/70) of DRG neurons, relative to $Ca_v2.3a$ expression in 2.8% (n=2/70) of DRG neurons. $Ca_v2.3e$ mRNA was mainly detected in small-sized neurons (n=12/14), but in only a few medium-sized neurons (n=2/14) and not in large-sized neurons, indicating the prominence of $Ca_v2.3e$ in nociceptive DRG neurons. Moreover, $Ca_v2.3e$ was preferentially expressed in tyrosine-kinase A (trkA)-positive, isolectin B4 (IB4)-negative and transient receptor potential vanilloid 1 (TRPV1)-positive neurons. These results suggest that $Ca_v2.3e$ may be the main R-type $Ca^{2+}$ channel isoform in nociceptive DRG neurons and thereby a potential target for pain treatment, not only in the trigeminal system but also in the spinal system.

Potential Involvement of p62, a Phosphotyrosine-independent Ligand of SH2 Domain of $p56^{lck}$, on UV-induced Apoptosis in Jurkat T-cell Line ($p56^{lck}$ SH2 domain 결합 단백질 p62가 Jurkat T-세포주의 세포예정사에 미치는 영향)

  • 정인실
    • Development and Reproduction
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    • v.2 no.2
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    • pp.165-171
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    • 1998
  • p62 is a novel cytoplsmic protein that binds to SH2 domain of p56$^{lck}$, lymphocyte-specific protein tyrosine kinase, and the expression of p62 was observed in most tissues. In addition p62 interacts with various proteins including ubiquitin and atypical PKC isoform, indicating its diverse biological role in different tissues. However, little is known about functional connection between p62 and its binding proteins. In the present study, a novel cellular protein, p62 has been shown to bind to 14-3-3 $\tau$ isoform that is specific for T cells. Moreover, overexpression of p62 in T cells caused to delay onset of UV-induced apoptosis characterized by DNA fragmentation and breakdown of poly (ADP-ribose) polymerase (PARP). Lately, 14-3-3 proteins have been shown to mediate survival signal via interacting proapoptotic Bad protein in the Iymphocyte. These results suggested the presence of p62-mediated regulatory mechanism during apoptosis in T cells, in which activation-induced apoptotic signal could be interfered by p62 and 14-3-3 protein.n.

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Interleukin-32 in Inflammatory Autoimmune Diseases

  • Kim, Soohyun
    • IMMUNE NETWORK
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    • v.14 no.3
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    • pp.123-127
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    • 2014
  • Interleukin-32 (IL-32) is a cytokine inducing crucial inflammatory cytokines such as tumor necrosis factor-${\alpha}(TNF{\alpha})$ and IL-6 and its expression is elevated in various inflammatory autoimmune diseases, certain cancers, as well as viral infections. IL-32 gene was first cloned from activated T cells, however IL-32 expression was also found in other immune cells and non-immune cells. IL-32 gene was identified in most mammals except rodents. It is transcribed as multiple-spliced variants in the absence of a specific activity of each isoform. IL-32 has been studied mostly in clinical fields such as infection, autoimmune, cancer, vascular disease, and pulmonary diseases. It is difficult to investigate the precise role of IL-32 in vivo due to the absence of IL-32 gene in mouse. The lack of mouse IL-32 gene restricts in vivo studies and restrains further development of IL-32 research in clinical applications although IL-32 new cytokine getting a spotlight as an immune regulatory molecule processing important roles in autoimmune, infection, and cancer. In this review, we discuss the regulation and function of IL-32 in inflammatory bowel diseases and rheumatoid arthritis.

Spatiotemporal expression of RCAN1 and its isoform RCAN1-4 in the mouse hippocampus after pilocarpine-induced status epilepticus

  • Cho, Kyung-Ok;Jeong, Kyoung Hoon;Cha, Jung-Ho;Kim, Seong Yun
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.1
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    • pp.81-88
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    • 2020
  • Regulator of calcineurin 1 (RCAN1) can be induced by an intracellular calcium increase and oxidative stress, which are characteristic features of temporal lobe epilepsy. Thus, we investigated the spatiotemporal expression and cellular localization of RCAN1 protein and mRNA in the mouse hippocampus after pilocarpine-induced status epilepticus (SE). Male C57BL/6 mice were given pilocarpine hydrochloride (280 mg/kg, i.p.) and allowed to develop 2 h of SE. Then the animals were given diazepam (10 mg/kg, i.p.) to stop the seizures and sacrificed at 1, 3, 7, 14, or 28 day after SE. Cresyl violet staining showed that pilocarpine-induced SE resulted in cell death in the CA1 and CA3 subfields of the hippocampus from 3 day after SE. RCAN1 immunoreactivity showed that RCAN1 was mainly expressed in neurons in the shammanipulated hippocampi. At 1 day after SE, RCAN1 expression became detected in hippocampal neuropils. However, RCAN1 signals were markedly enhanced in cells with stellate morphology at 3 and 7 day after SE, which were confirmed to be reactive astrocytes, but not microglia by double immunofluorescence. In addition, realtime reverse transcriptase-polymerase chain reaction showed a significant upregulation of RCAN1 isoform 4 (RCAN1-4) mRNA in the SE-induced hippocampi. Finally, in situ hybridization with immunohistochemistry revealed astrocytic expression of RCAN1-4 after SE. These results demonstrate astrocytic upregulation of RCAN1 and RCAN1-4 in the mouse hippocampus in the acute and subacute phases of epileptogenesis, providing foundational information for the potential role of RCAN1 in reactive astrocytes during epileptogenesis.

EXPRESSION AND FUNCTIONAL CHARACTERIZATION OF AMINO ACID TRANSPORT SYSTEM L IN SAOS2 HUMAN OSTEOGENIC SARCOMA CELLS (사람 골육종 세포 Saos2에서 아미노산 수송계 L의 발현 및 기능적 특성)

  • Kim, Su-Gwan;Kim, Hyun-Ho;Kim, Chang-Hyun;Kim, Do-Kyung
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.3
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    • pp.200-208
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    • 2006
  • Amino acids are required for protein synthesis and energy sources in all living cells. The amino acid transport system L is a major nutrient transport system that is responsible for $Na^+$-independent transport of neutral amino acids including several essential amino acids. In malignant tumors, the L-type amino acid transporter 1 (LAT1), the first isoform of system L, is highly expressed to support tumor cell growth. In the present study, the expression and functional characterization of amino acid transport system L were, therefore, investigated in Saos2 human osteogenic sarcoma cells. RT-PCR and western blot analyses have revealed that the Saos2 cells expressed the LAT1 and the L-type amino acid transporter 2 (LAT2), the second isoform of system L, together with their associating protein heavy chain of 4F2 antigen (4F2hc) in the plasma membrane, but the expression of LAT2 was very weak. The uptakes of [${14}^C$]L-leucine by Saos2 cells were $Na^+$-independent and were completely inhibited by the system L selective inhibitor, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). The affinity of [${14}^C$]L-leucine uptake and the inhibition profiles of [${14}^C$]L-leucine uptake by various amino acids in the Saos2 cells were comparable with those for the LAT1 expressed in Xenopus oocytes. The majority of [${14}^C$]L-leucine uptake is, therefore, mediated by LAT1 in the Saos2 cells. These results suggest that the transports of neutral amino acids including several essential amino acids into Saos2 human osteogenic sarcoma cells are for the most part mediated by LAT1. Therefore, the Saos2 human osteogenic sarcoma cells are excellent tools for examine the properties of LAT1. Moreover, the specific inhibition of LAT1 in tumor cells might be a new rationale for anti-tumor therapy.

Role of Lysyl Oxidase Family during Odontoblastic Differentiation of Human Dental Pulp Cells Induced with Odontogenic Supplement (인간치수세포에서 상아모세포의 분화과정 동안 Lysyl Oxidase Family의 역할)

  • Lee, Hwa-Jeong;Han, Soo-Yeon
    • Journal of dental hygiene science
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    • v.13 no.3
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    • pp.296-303
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    • 2013
  • Lysyl oxidase (LOX), extracellular matrix enzyme, is catalyzing lysine-derived crosslinks in collagen and elastin. Recently, several LOX-like proteins (LOXL, LOXL2, LOXL3 and LOXL4) have been identified in human but their specific functions are still largely unknown. The purpose of this study was to evaluate the function of the LOX family genes during odontoblastic differentiation of human dental pulp (HDP) cells induced with odontogenic supplement (OS). The messenger RNA (mRNA) expression of LOX family genes and differentiation markers was assessed by reverse transcriptase polymerase chain reaction analysis (RT-PCR). The formation of mineralization nodules was evaluated by alrizarin red S staining. Amine oxidase activity of HDP cells was measured by peroxidase-coupled fluormetric assay. The expressions of differentiation markers, such as alkaline phosphatase (ALP), osteopontin (OPN), osteocalcin (OCN), dentin matrix protein1 (DMP1), dentin sialophosphoprotein (DSPP) in HDP cells were increased after treatment with OS media. The LOX and LOXL mRNA expression were gradually increased in OS media, whereas LOX enzyme activities were markedly detected on day 7. The mRNA expression and LOX enzyme activity of collagen type I was very similar to the pattern of LOX gene. In this study, the expression of LOX and its isoforms, and activity of LOX were highly regulated during odontoblastic differentiation. Thus, these results suggest that LOX plays a key role in odontoblastic differentiation of HDP cells.

Relationships between Myosin Light Chain Isoforms, Muscle Fiber Characteristics, and Meat Quality Traits in Porcine Longissimus Muscle

  • Choi, Young-Min;Ryu, Youn-Chul;Lee, Sang-Hoon;Kim, Byoung-Chul
    • Food Science and Biotechnology
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    • v.14 no.5
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    • pp.639-644
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    • 2005
  • The aim of this study was to investigate the effect of the myosin light chain (MLC) isoforms on the muscle fiber characteristics and meat quality traits in porcine longissimus muscle. Pale, soft, exudative (PSE) samples had a lower content of essential light chain (ELC) 1S isoforms and a higher proportion of the fiber type IIB than the reddish-pink, firm, non-exudative (RFN) samples. These compositions suggest that the PSE pork has a higher glycolytic and a lower oxidative capacity than the RFN pork. Therefore, these characteristics of PSE pork might affect the metabolic rate and meat quality traits, including protein solubility. In addition, the indicator traits of the postmortem metabolic rate were related to the ELC 1F/3F ratio ($pH_{45\;min}$: r = -0.43, P < 0.001; R-value: r = 0.53, P < 0.001). These results suggest that the MLC isoform composition can affect the postmortem metabolic rate and meat quality traits.