Phospholipase C-β1 mediates α1-adrenergic receptor-stimulated activation of the sodium–hydrogen exchanger in Chinese hamster lung fibroblasts (CCL39)

2005 ◽  
Vol 83 (2) ◽  
pp. 123-132 ◽  
Author(s):  
J J Provost ◽  
S M Olmschenk ◽  
A L Metcalf ◽  
N Korpi ◽  
H Thronson ◽  
...  
Keyword(s):  
Phospholipase C ◽  
Cell Lines ◽  
Adrenergic Receptor ◽  
Chinese Hamster ◽  
Lung Fibroblasts ◽  
Specific Chemical ◽  
Sodium Hydrogen ◽  
Sodium Hydrogen Exchanger ◽  
Nhe1 Activity ◽  
Ccl39 Cells

The activation of the Na+–H+ exchanger 1 (NHE1) and extracellular-signal regulated kinase (ERK) phospho rylation in Chinese hamster lung fibroblasts (CCL39) was characterized in response to the specific α1-adrenergic agonist, phenylephrine (PE). Addition of 100 µmol PE/L increased the steady-state intracellular pH (pHi) by 0.16 ± 0.03 pH units, as well as increasing the phosphorylation of ERK. The response of NHE1 to PE in CCL39 cells was determined by the use of specific antagonists. Use of 2 specific chemical inhibitors of phosphoinositide-specific phospholipase C (PLC) reduced the ability of PE to activate either the exchanger or ERK. Studies were conducted in PLCβ-deficient cell lines derived from parental CCL39 cells. NHE1 activity in both mutant cell lines was increased in response to phorbal esters or lysophosphatidic acid, whereas the addition of PE only caused a minimal change in either pHi or ERK phosphorylation. These results, combined with reconstitution experiments with exogenously expressed PLCβ1, PLCβ2, or PLCβ3, revealed that stimulation of NHE1 activity by PE in CCL39 cells is a PLCβ1-coupled event. Furthermore, the data indicate that α1-adrenergic signaling of PLCβ is upstream of ERK activation. These data demonstrate that PLCβ1 is primarily involved in the activation of NHE1 in CCL39 fibroblasts.Key words: CCL39, sodium hydrogen exchanger, ERK, α1-adrenergic receptor, phospholipase Cβ.

scholarly journals Alpha 2-adrenergic agonists stimulate DNA synthesis in Chinese hamster lung fibroblasts transfected with a human alpha 2-adrenergic receptor gene.

1990 ◽  
Vol 1 (6) ◽  
pp. 445-451 ◽  
Author(s):  
K Seuwen ◽  
I Magnaldo ◽  
B K Kobilka ◽  
M G Caron ◽  
J W Regan ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Growth Factor ◽  
Dna Synthesis ◽  
Adrenergic Receptors ◽  
Adrenergic Receptor ◽  
Inositol Phosphates ◽  
Receptor Gene ◽  
Chinese Hamster ◽  
Lung Fibroblasts ◽  
Ccl39 Cells

To test the hypothesis that agents activating receptors negatively coupled to adenylyl cyclase (AC) can stimulate cell proliferation, we have expressed a human alpha 2-adrenergic receptor (alpha 2-C10) in CCL39 cells and studied the effects of alpha 2-agonists on reinitiation of DNA synthesis in quiescent cells. We report that the alpha 2-agonists epinephrine and clonidine stimulate [3H]-thymidine incorporation in synergy with fibroblast growth factor and that the alpha 2-antagonist yohimbine efficiently inhibits this response. Epinephrine- and clonidine-stimulated DNA synthesis is completely blocked by pertussis toxin and correlates well with the inhibition of prostaglandin E1-stimulated AC. Thus, their action closely resembles the action of serotonin in the same cell system, which is mediated through 5-HT1b receptors. In fact, serotonin- and epinephrine-stimulated DNA synthesis reinitiation is not additive, suggesting that both agents act through a common pathway. Interestingly, alpha 2-agonists also induced a moderate release of inositol phosphates, indicating that alpha 2-adrenergic receptors can interact both with the AC and phospholipase C messenger system. Activation of phosphoinositide (PI) turnover by epinephrine leads to a significant stimulation of Na+/H+ exchange but is insufficient to trigger a mitogenic response in CCL39 cells, as will be discussed. We found no evidence for epinephrine-induced activation of Na+/H+ exchange by a mechanism independent of PI breakdown.Our data show that alpha 2-adrenergic receptors can play a role in the regulation of cell proliferation in an appropriate context; also, the data support the hypothesis that receptors negatively coupled to AC must be taken into account as mediators of growth factor action in fibroblasts, in particular when activated in parallel with receptor tyrosine kinases.

scholarly journals Investigation of the specific α1‐adrenergic receptor pathway link to phosphorylation of sodium‐hydrogen exchanger by RhoA kinase

2007 ◽  
Vol 21 (6) ◽  
Author(s):  
David Luther Mork ◽  
Lisa Streitz ◽  
Robert Reames ◽  
Moses Wananu ◽  
Mark Wallert ◽  
...  
Keyword(s):  
Adrenergic Receptor ◽  
Sodium Hydrogen ◽  
Rhoa Kinase ◽  
Sodium Hydrogen Exchanger

Phosphorylation and Palmitoylation Dynamically Regulate Sodium Hydrogen Exchanger Isoform 1 (NHE1) Activity: Implications in Health and Disease

2020 ◽  
Vol 34 (S1) ◽  
pp. 1-1
Author(s):  
Moriah J. Hovde ◽  
Danielle E. Bolland ◽  
Clair Bakker ◽  
Taylor Cottle ◽  
Mark A. Wallert ◽  
...  
Keyword(s):  
Sodium Hydrogen ◽  
Sodium Hydrogen Exchanger ◽  
Nhe1 Activity ◽  
Health And Disease

scholarly journals Alkaline Cytosolic pH and High Sodium Hydrogen Exchanger 1 (NHE1) Activity in Th9 Cells

2016 ◽  
Vol 291 (45) ◽  
pp. 23662-23671 ◽  
Author(s):  
Yogesh Singh ◽  
Yuetao Zhou ◽  
Xiaolong Shi ◽  
Shaqiu Zhang ◽  
Anja T. Umbach ◽  
...  
Keyword(s):  
Cytosolic Ph ◽  
High Sodium ◽  
Sodium Hydrogen ◽  
Th9 Cells ◽  
Sodium Hydrogen Exchanger ◽  
Nhe1 Activity

scholarly journals Synthesis and anticancer effects of conjugates of doxorubicin and unsaturated fatty acids (LNA and DHA)

2019 ◽  
Vol 28 (12) ◽  
pp. 2153-2164 ◽  
Author(s):  
Magdalena Mielczarek-Puta ◽  
Marta Struga ◽  
Piotr Roszkowski
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Tumor Cells ◽  
Cell Lines ◽  
Unsaturated Fatty Acids ◽  
Chinese Hamster ◽  
Proton Nuclear Magnetic Resonance ◽  
Lung Fibroblasts ◽  
Amide Derivatives ◽  
Nuclear Magnetic

Abstract Doxorubicin (DOX) is a leading cytostatic drug with many adverse effects in use. We are still looking for methods that will allow us to preserve the therapeutic effect against the tumor cells and reduce the toxicity to the normal cells. In our work, we obtained amide derivatives of DOX by reaction of the amino group with α-linolenic (LNA) and docosahexaenoic (DHA) acids (2, 3), as well as double-substituted derivatives via amide and ester linkages (4, 5). The structures of the compounds were confirmed by Proton Nuclear Magnetic Resonance (1H NMR), Carbon-13 Nuclear Magnetic Resonance (13C NMR), and High Resolution Mass Spectrometry (HRMS) analyses. For all compounds 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cytotoxic effect on human cancer cell lines (SW480, SW620, and PC3) and Chinese hamster lung fibroblasts (V79) that were used as a control. The cytotoxic activity was established by calculation of the inhibitory concentration IC50. In addition, a cytotoxic capacity against tumor cells for tested compounds was expressed as a selectivity factor (selectivity index, SI). Lactate dehydrogenase (LDH) assay was performed for all compounds to assess the level of cell damage. To explain the basic mechanism of cell death induction the Annexin V-FITC/IP flow cytometry analysis was investigated. We found that all studied conjugates exhibit lower cytotoxicity but higher selectivity than DOX. Among the all derivatives, the conjugates formed by the amide and ester linkages (4, 5) were found to be more promising compared with conjugates (2, 3) formed only by the amide linkage. They show high cytotoxicity toward the tumor cell lines and moderate cytotoxicity towards the normal cell line.

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