1.Effect of lysophosphatidic acid on differentiation of embryonic neural stem cells into neuroglial cells in rats in vitro.
Acta Physiologica Sinica 2007;59(6):759-764
To study the effect of lysophosphatidic acid (LPA) on the differentiation of embryonic neural stem cells (NSCs) into neuroglial cells in rats in vitro, both oligodendrocytes and astrocytes were detected by their marker proteins galactocerebroside (Gal-C) and glial fibrillary acidic protein (GFAP), respectively, using double-labeling immunocytochemistry. RT-PCR assay was also used for analyzing the expression of LPA receptors in NSCs. Our results showed that: (1) LPA at different concentrations (0.01-3.0 mumol/L) was added to culture medium and cell counting was carried out on the 7th day in all groups. Exposure to LPA led to a dose-dependent increase of oligodendrocytes with the response peaked at 1.0 mumol/L, with an increased percentage of 32.6% (P<0.01) of total cells as compared to that of 8.5% in the vehicle group. (2) LPA showed no effect on the differentiation of NSCs into astrocytes. (3) RT-PCR assay showed that LPA(1) and LPA(3) receptors were strongly expressed while LPA(2) receptor expressed weakly in NSCs. These results suggest that LPA at low concentration might act as an extracellular signal through the receptors in NSCs, mainly LPA(1) and LPA(3) receptors, to promote the differentiation of NSCs into oligodendrocytes, while it exhibits little, if any, conceivable effect on the differentiation of NSCs into astrocytes.
Animals
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Cell Differentiation
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drug effects
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Cells, Cultured
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Lysophospholipids
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pharmacology
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Neural Stem Cells
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cytology
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drug effects
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Neuroglia
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cytology
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Rats
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Receptors, Lysophosphatidic Acid
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metabolism
2.Synovial fluid of patients with rheumatoid arthritis induces alpha-smooth muscle actin in human adipose tissue-derived mesenchymal stem cells through a TGF-beta1-dependent mechanism.
Hae Young SONG ; Min Young KIM ; Kyung Hye KIM ; Il Hwan LEE ; Sang Hun SHIN ; Jung Sub LEE ; Jae Ho KIM
Experimental & Molecular Medicine 2010;42(8):565-573
Rheumatoid arthritis (RA) is a chronic, inflammatory autoimmune disorder that causes the immune system to attack the joints. Transforming growth factor-beta1 (TGF-beta1) is a secreted protein that promotes differentiation of synovial fibroblasts to alpha-smooth muscle actin (alpha-SMA)-positive myofibroblasts to repair the damaged joints. Synovial fluid from patients with RA (RA-SF) induced expression of alpha-SMA in human adipose tissue-derived mesenchymal stem cells (hASCs). RA-SF-induced alpha-SMA expression was abrogated by immunodepletion of TGF-beta1 from RA-SF with anti-TGF-beta1 antibody. Furthermore, pretreatment of hASCs with the TGF-beta type I receptor inhibitor SB431542 or lentiviral small hairpin RNA-mediated silencing of TGF-beta type I receptor expression in hASCs blocked RA-SF-induced alpha-SMA expression. Small interfering RNA-mediated silencing of Smad2 or adenoviral overexpression of Smad7 (an inhibitory Smad isoform) completely inhibited RA-SF-stimulated alpha-SMA expression. These results suggest that TGF-beta1 plays a pivotal role in RA-SF-induced differentiation of hASCs to alpha-SMA-positive cells.
Actins/*metabolism
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Adipose Tissue/*cytology
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Arthritis, Rheumatoid/*metabolism
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Humans
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Mesenchymal Stem Cells/*metabolism
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Receptors, Lysophosphatidic Acid/metabolism
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Signal Transduction
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Smad2 Protein/metabolism
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Stress Fibers/metabolism
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Synovial Fluid/*metabolism
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Transforming Growth Factor beta1/*metabolism
3.Role of LPA and the Hippo pathway on apoptosis in salivary gland epithelial cells.
Sung Min HWANG ; MeiHong JIN ; Yong Hwan SHIN ; Seul KI CHOI ; Eun NAMKOONG ; MinKyoung KIM ; Moon Yong PARK ; Kyungpyo PARK
Experimental & Molecular Medicine 2014;46(12):e125-
Lysophosphatidic acid (LPA) is a bioactive lysophospholipid involved in numerous physiological responses. However, the expression of LPA receptors and the role of the Hippo signaling pathway in epithelial cells have remained elusive. In this experiment, we studied the functional expression of LPA receptors and the associated signaling pathway using reverse transcriptase-PCR, microspectrofluorimetry, western blotting and immunocytochemistry in salivary gland epithelial cells. We found that LPA receptors are functionally expressed and involved in activating the Hippo pathway mediated by YAP/TAZ through Lats/Mob1 and RhoA/ROCK. Upregulation of YAP/TAZ-dependent target genes, including CTGF, ANKRD1 and CYR61, has also been observed in LPA-treated cells. In addition, based on data suggesting that tumor necrosis factor (TNF)-alpha induces cell apoptosis, LPA upregulates TNF-induced caspase-3 and cleaved Poly(ADP-ribose)polymerase (PARP). However, small interfering RNA treatment to Yes-associated protein (YAP) or transcriptional co-activator with a PDZ-binding motif (TAZ) significantly decreased TNF-alpha- and LPA-induced apoptosis, suggesting that YAP and TAZ modulate the apoptotic pathway in salivary epithelial cells.
Adaptor Proteins, Signal Transducing/genetics/metabolism
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*Apoptosis
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Cell Line
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Epithelial Cells/*cytology/metabolism
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Gene Expression Regulation
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Humans
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Intracellular Signaling Peptides and Proteins/genetics/metabolism
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Lysophospholipids/*metabolism
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Phosphoproteins/genetics/metabolism
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Protein-Serine-Threonine Kinases/*metabolism
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RNA Interference
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RNA, Small Interfering/genetics
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Receptors, Lysophosphatidic Acid/genetics/*metabolism
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Salivary Glands/*cytology/metabolism
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*Signal Transduction
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Tumor Necrosis Factor-alpha/metabolism
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rho-Associated Kinases/metabolism
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rhoA GTP-Binding Protein/metabolism
4.Lysophosphatidic acid receptor 2 and Gi/Src pathway mediate cell motility through cyclooxygenase 2 expression in CAOV-3 ovarian cancer cells.
Kang Jin JEONG ; Soon Young PARK ; Ji Hye SEO ; Kyung Bok LEE ; Wahn Soo CHOI ; Jeung Whan HAN ; Jae Ku KANG ; Chang Gyo PARK ; Yong Kee KIM ; Hoi Young LEE
Experimental & Molecular Medicine 2008;40(6):607-616
Lysophosphatidic acid (LPA) is a bioactive phospholipids and involves in various cellular events, including tumor cell migration. In the present study, we investigated LPA receptor and its transactivation to EGFR for cyclooxygenase-2 (COX-2) expression and cell migration in CAOV-3 ovarian cancer cells. LPA induced COX-2 expression in a dose-dependent manner, and pretreatment of the cells with pharmacological inhibitors of Gi (pertussis toxin), Src (PP2), EGF receptor (EGFR) (AG1478), ERK (PD98059) significantly inhibited LPA- induced COX-2 expression. Consistent to these results, transfection of the cells with selective Src siRNA attenuated COX-2 expression by LPA. LPA stimulated CAOV-3 cell migration that was abrogated by pharmacological inhibitors and antibody of EP2. Higher expression of LPA2 mRNA was observed in CAOV-3 cells, and transfection of the cells with a selective LPA2 siRNA significantly inhibited LPA-induced activation of EGFR and ERK, as well as COX-2 expression. Importantly, LPA2 siRNA also blocked LPA-induced ovarian cancer cell migration. Collectively, our results clearly show the significance of LPA2 and Gi/Src pathway for LPA-induced COX-2 expression and cell migration that could be a promising drug target for ovarian cancer cell metastasis.
Butadienes/pharmacology
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Cell Line, Tumor
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Cell Movement/drug effects/*physiology
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Cyclooxygenase 2/*biosynthesis
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Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors/metabolism
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Female
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Flavonoids/pharmacology
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GTP-Binding Protein alpha Subunits, Gi-Go/antagonists & inhibitors/*metabolism
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Humans
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Lysophospholipids/pharmacology
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Nitriles/pharmacology
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Ovarian Neoplasms/metabolism/*pathology
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Pertussis Toxin/pharmacology
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Protein-Tyrosine Kinases/antagonists & inhibitors/*metabolism
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Proto-Oncogene Proteins/antagonists & inhibitors/*metabolism
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Pyrimidines/pharmacology
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Receptor, Epidermal Growth Factor/antagonists & inhibitors/metabolism
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Receptors, Lysophosphatidic Acid/*metabolism
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Receptors, Prostaglandin E/metabolism
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Signal Transduction
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Transcriptional Activation
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Tyrphostins/pharmacology