Econazole induces increases in free intracellular Ca2+ concentrations in human osteosarcoma cells

2005 ◽  
Vol 24 (9) ◽  
pp. 453-458 ◽  
Author(s):  
H-T Chang ◽  
C-S Liu ◽  
C-T Chou ◽  
C-H Hsieh ◽  
C-H Chang ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Phospholipase C ◽  
Dependent Manner ◽  
Osteosarcoma Cells ◽  
Independent Manner ◽  
Concentration Dependent Manner ◽  
Human Osteosarcoma Cells ◽  
Intracellular Ca ◽  
In Vitro Effects

Econazole is an antifungal drug with different in vitro effects. However, econazole's effect on osteoblast like cells is unknown. In human MG63 osteosarcoma cells, the effect of econazole on intracellular Ca2+ concentrations ([Ca2+]i) was explored by using fura-2. At a concentration of 0.1 μM, econazole started to cause a rise in [Ca2+]i in a concentration-dependent manner. Econazole-induced [Ca2+]i rise was reduced by 74% by removal of extracellular Ca2+. The econazole-induced Ca2+ influx was mediated via a nimodipine-sensitive pathway. In Ca2+ free medium, thapsigargin, an inhibitor of the endoplasmic reticulum Ca2+ ATPase, caused a [Ca2+]i rise, after which the increasing effect of econazole on [Ca2+]i was abolished. Pretreatment of cells with econazole to deplete Ca2+ stores totally prevented thapsigargin from releasing Ca2+. U73122, an inhibitor of phospholipase C, abolished histamine (an inositol 1,4,5-trisphosphate dependent Ca2+ mobilizer)-induced, but not econazoleinduced, [Ca2+]i rise. Econazole inhibited 76% of thapsigargin-induced store-operated Ca2+ entry. These findings suggest that in MG63 osteosarcoma cells, econazole increases [Ca2+]i by stimulating Ca2+ influx and Ca2+ release from the endoplasmic reticulum via a phospholipase C-independent manner. In contrast, econazole acts as a potent blocker of store-operated Ca2+ entry.

Carvedilol-induced elevation in cytosolic free Ca2+ level and apoptosis in SIRC corneal epithelial cells

2009 ◽  
Vol 29 (6) ◽  
pp. 477-487
Author(s):  
Pochuen Shieh ◽  
Chih-Hung Lee ◽  
Ng Ling Yi ◽  
Chung-Ren Jan
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Epithelial Cells ◽  
Phospholipase C ◽  
Dependent Manner ◽  
Corneal Epithelial Cells ◽  
Concentration Dependent Manner ◽  
Corneal Epithelial ◽  
Kinase C

The effect of the cardiovascular drug carvedilol on cytosolic free Ca2+ concentrations ([Ca 2+]i) and viability was examined in Statens Seruminstitut rabbit cornea (SIRC) corneal epithelial cells. [Ca2+]i and cell viability were measured using the fluorescent dyes fura-2 and 4-[3-[4-lodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] (WST-1), respectively. Carvedilol at concentrations between 1 and 30 μM increased [Ca 2+]i in a concentration-dependent manner. The Ca2+ signal was reduced partly by removing extracellular Ca2+. Carvedilol induced Mn2+ quench of fura-2 fluorescence implicating Ca2+ influx. The Ca2+ influx was inhibited by suppression of protein kinase C activity. In Ca2+-free medium, after pretreatment with 1 μM thapsigargin (an endoplasmic reticulum Ca 2+ pump inhibitor), carvedilol-induced [Ca2+]i rise was reduced; and conversely, carvedilol pretreatment inhibited a major part of thapsigargin-induced [Ca 2+]i rise. Addition of the phospholipase C inhibitor 1-[6-[[17 beta-3-methoxyestra-1,3,5(10)-trien-17-yl]amino] hexyl]-1H-pyrrole-2,5-dione (U73122; 2 μM) did not change carvedilol-induced [Ca2+]i rise. At concentrations between 5 and 70 μM, carvedilol killed cells in a concentration-dependent manner. The cytotoxic effect of 20 μM carvedilol was not reversed by prechelating cytosolic Ca2+ with BAPTA/AM. Apoptosis was induced by 5—70 μM carvedilol. Collectively, in SIRC corneal epithelial cells, carvedilol-induced [Ca2+]i rises by causing Ca2+ release from the endoplasmic reticulum in a phospholipase C-independent manner, and Ca 2+ influx via protein kinase C-regulated Ca2+ channels. Carvedilol-caused cytotoxicity was mediated by Ca2+-independent apoptosis in a concentration-dependent manner.

scholarly journals Effect of thimerosal on Ca2+ movement and viability in human oral cancer cells

2009 ◽  
Vol 28 (5) ◽  
pp. 301-308 ◽  
Author(s):  
LN Kuo ◽  
CJ Huang ◽  
YC Fang ◽  
CC Huang ◽  
JL Wang ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Oral Cancer ◽  
Cancer Cells ◽  
Phospholipase C ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Flow Cytometry Data ◽  
Entry Inhibitors ◽  
Oral Cancer Cells ◽  
Induced Apoptosis

The effect of thimerosal on cytosolic free Ca2+ concentrations ([Ca2+]i ) in human oral cancer cells (OC2) is unclear. This study explored whether thimerosal changed basal [Ca2+]i levels in suspended OC2 cells using fura-2. Thimerosal at concentrations between 1and 50 μM increased [Ca2+]i in a concentration-dependent manner. The Ca2+ signal was reduced partly by removing extracellular Ca 2+. Thimerosal-induced Ca2+ influx was not blocked by L-type Ca2+ entry inhibitors and protein kinase C modulators (phorbol 12-myristate 13-acetate [PMA] and GF109203X). In Ca2+-free medium, 50 μM thimerosal failed to induce a [Ca2+]i rise after pretreatment with thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor). Inhibition of phospholipase C with U73122 did not change thimerosal-induced [Ca2+]i rises. At concentrations between 5 and 10 μM, thimerosal killed cells in a concentration-dependent manner. The cytotoxic effect of 8 μM thimerosal was potentiated by prechelating cytosolic Ca2+ with the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetate/acetomethyl (BAPTA/ AM). Flow cytometry data suggested that 1—7 μM thimerosal-induced apoptosis in a concentration-dependent manner. Collectively, in OC2 cells, thimerosal-induced [Ca2+]i rises by causing phospholipase C-independent Ca2+ release from the endoplasmic reticulum and Ca2+ influx through non—L-type Ca2+ channels. Thimerosal killed cells in a concentration-dependent manner through apoptosis.

Carvacrol Suppresses Human Osteosarcoma Cells via the Wnt/β-Catenin Signaling Pathway

2021 ◽  
Vol 21 ◽  
Author(s):  
Songou Zhang ◽  
Lei He ◽  
Jinxiang Shang ◽  
Long Chen ◽  
Yifan Xu ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Colony Formation ◽  
Apoptotic Cell ◽  
Chinese Herbs ◽  
Migration And Invasion ◽  
Osteosarcoma Cell ◽  
Dependent Manner ◽  
Osteosarcoma Cells ◽  
Concentration Dependent Manner ◽  
Human Osteosarcoma Cells

Background: Carvacrol is a monoterpenic phenol extracted from traditional Chinese herbs, including oregano and thyme. Currently, carvacrol has been widely studied for its therapeutic role in central nervous system diseases, liver diseases and digestive system cancer. Objective: However, the role of carvacrol in osteosarcoma and its underlying molecular mechanism remain elusive. Here, we aimed to examine the anticancer effects of carvacrol on osteosarcoma. Methods: The effects of carvacrol on the osteosarcoma proliferation capacity were revealed by CCK-8 and colony formation assays. Flow cytometry and Hoechst assays were used to determine the effects of carvacrol on osteosarcoma cell apoptosis. The effect of carvacrol on migration and invasion of osteosarcoma cells was determined by wound healing and transwell tests. Protein expression was evaluated by WB assays. The suppressive effects of carvacrol on osteosarcoma in vivo were examined by a xenograft animal model, immunohistochemistry and HE staining. Results: We demonstrated that carvacrol treatment reduced viability and inhibited the colony formation of U2OS and 143B cells in a concentration-dependent manner. Apoptotic cell number increased after exposure to carvacrol. Meanwhile, the expression of Bax increased, and that of Bcl-2 decreased by carvacrol treatment. In addition, the MMP-9 expression and migration and invasion of 143B and U2OS cells were inhibited by carvacrol. We also found that these carvacrol-induced effects on osteosarcoma are associated with the regulation of the Wnt/β-catenin signaling pathway. Conclusion: Our findings suggest that carvacrol suppresses proliferation, migration, invasion and promotes apoptosis in osteosarcoma cells, in part by regulating the Wnt/β-catenin signaling pathway.

scholarly journals G-protein βγ subunits antagonize protein kinase C-dependent phosphorylation and inhibition of phospholipase C-β1

1997 ◽  
Vol 326 (3) ◽  
pp. 701-707 ◽  
Author(s):  
Irene LITOSCH
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phospholipase C ◽  
G Protein ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Negative Feedback Regulation ◽  
Kinase C ◽  
Stimulation Of

Protein kinase C (PKC) isoforms phosphorylated phospholipase C-β1 (PLC-β1) in vitro as follows: PKCα ≫ PKCϵ; not PKCζ. PLC-β3 was not phosphorylated by PKCα. G-protein βγ subunits inhibited the PKCα phosphorylation of PLC-β1 in a concentration-dependent manner. Half-maximal inhibition occurred with 500 nM βγ. G-protein βγ subunits also antagonized the PKCα-mediated inhibition of PLC-β1 enzymic activity. PKCα, in turn, inhibited the stimulation of PLC-β1 activity by βγ. There was little effect of PKCα on the stimulation of PLC-β1 by αq/11–guanosine 5′[γ-thio]triphosphate (GTP[S]). These findings demonstrate that G protein βγ subunits antagonize PKCα regulation of PLC-β1. Thus βγ subunits might have a role in modulating the negative feedback regulation of this signalling system by PKC.

scholarly journals Antiadipogenic Effects ofAster glehniExtract: In Vivo and In Vitro Effects

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Heon-Myung Lee ◽  
Gabsik Yang ◽  
Tae-Gue Ahn ◽  
Myung-Dong Kim ◽  
Agung Nugroho ◽  
...  
Keyword(s):  
Therapeutic Potential ◽  
Control Diet ◽  
Epididymal Adipose Tissue ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Master Regulators ◽  
In Vitro Effects

Aster glehni(AG) is a Korean traditional herb that grows in Ulleungdo Island, Republic of Korea. None of the several reports on AG include a determination of the effect of AG on adipogenesis. The primary aim of this study was to determine whether AG attenuates adipogenesis in mouse 3T3-L1 cells and epididymal fat tissue. AG blocked the differentiation of 3T3-L1 preadipocytes in a concentration-dependent manner and suppressed the expression of adipogenesis-related genes such asPPARγ,C/EBPα, andSREBP1c, the master regulators of adipogenesis. Male C57BL/6J mice were divided randomly and equally into 4 diet groups: control diet (CON), high-fat diet (HFD), HFD with 1% AG extract added (AG1), and HFD with 5% AG extract added (AG5). The experimental animals were fed HFD and the 2 combinations for 10 weeks. Mice fed HFD with AG gained less body weight and visceral fat-pad weight than did the mice fed HFD alone. Moreover, AG inhibited the expression of important adipogenic genes such asPPARγ,C/EBPα,SREBP1c,LXR, and leptin in the epididymal adipose tissue of the mice treated with AG1 and AG5. These findings indicate antiadipogenic and antiobesity effects of AG and suggest its therapeutic potential in obesity and obesity-related diseases.

Effect of Riluzole on Ca2+ Movement and Cytotoxicity in Madin-Darby Canine Kidney Cells

2006 ◽  
Vol 25 (8) ◽  
pp. 461-469 ◽  
Author(s):  
W-C Chen ◽  
H-H Cheng ◽  
C-J Huang ◽  
C-T Chou ◽  
S-I Liu ◽  
...  
Keyword(s):  
Mdck Cells ◽  
Dependent Manner ◽  
Madin Darby Canine Kidney ◽  
Concentration Dependent Manner ◽  
Intracellular Ca ◽  
Pre Treatment ◽  
Lateral Sclerosis ◽  
Darby Canine Kidney ◽  
Canine Kidney

Riluzole is a drug used in the treatment of amyotrophic lateral sclerosis; however, its in vitro action is unclear. In this study, the effect of riluzole on intracellular Ca2- concentration ([Ca2-]i) in Madin-Darby canine kidney (MDCK) cells was investigated using the Ca2--sensitive fluorescent dye, fura-2. Riluzole (100 -500 mM) caused a rapid and sustained increase of [Ca2-]i in a concentration-dependent manner (EC50 = 150 mM). Some 40 and 50% of this [Ca2-]i increase was prevented by the removal of extracellular Ca2- and the addition of La3-, respectively, but was unchanged by dihydropyridines, verapamil and diltiazem. In Ca2--free medium, thapsigargin -an inhibitor of the endoplasmic reticulum (ER) Ca2--ATPase -caused a monophasic [Ca2-]i increase, after which the increasing effect of riluzole on [Ca2-]iwas attenuated by 70%; in addition, pre-treatment with riluzole abolished thapsigargin-induced [Ca2-]i increases. U73122, an inhibitor of phospholipase C (PLC), abolished ATP (but not riluzole)-induced [Ca2-]i increases. At concentrations of 250 and 500 mM, riluzole killed 40 and 95% cells, respectively. The cytotoxic effect of riluzole (250 mM) was unaltered by pre-chelating cytosolic Ca2- with BAPTA. Collectively, in MDCK cells, riluzole rapidly increased [Ca2-]i by stimulating extra-cellular Ca2- influx via an La3--sensitive pathway and intracellular Ca2- release from the ER via, as yet, unidentified mechanisms. Furthermore, riluzole caused Ca2--unrelated cytotoxicity in a concentration-depen-dent manner.

scholarly journals Capacitative calcium entry is involved in steroidogenesis in bovine adrenocortical fasciculata cells

2000 ◽  
Vol 167 (3) ◽  
pp. 473-478 ◽  
Author(s):  
T Ebisawa ◽  
I Kondo ◽  
E Masaki ◽  
S Hori ◽  
M Kawamura
Keyword(s):  
Endoplasmic Reticulum ◽  
Incubation Medium ◽  
Dependent Manner ◽  
Atpase Inhibitor ◽  
Capacitative Calcium Entry ◽  
Concentration Dependent Manner ◽  
Glass Coverslip ◽  
Fura 2 ◽  
Intracellular Ca ◽  
Sustained Increase

Capacitative Ca(2+) entry into bovine adrenocortical fasciculata cells was investigated by using the mobilization of intracellular Ca(2+) concentration ([Ca(2+)](i)) and Ca(2+)-induced steroidogenesis as the indicators. Bovine adrenocortical fasciculata cells on a glass coverslip were loaded with fura-2. The [Ca(2+)](i) mobilization was detected by a change of fura-2 fluorescence intensity. In the intracellular Ca(2+) store depleted cells, the addition of Ca(2+) to the incubation medium elicited a marked and sustained increase in [Ca(2+)](i). In the intracellular Ca(2+) store non-depleted cells, the addition of thapsigargin, an endoplasmic reticulum Ca(2+)-ATPase inhibitor, in the absence of extracellular Ca(2+), induced a slight and transient increase in [Ca(2+)](i), but an extensive and sustained increase in [Ca(2+)](i) was obtained by adding Ca(2+) to the incubation medium after the thapsigargin treatment. The sustained increase induced by thapsigargin was not inhibited by nifedipine, but was inhibited by Zn(2+) and Cd(2+) in a concentration-dependent manner. The effect of Zn(2+) was more potent than that of Cd(2+). Thapsigargin stimulated steroidogenesis in the presence of extracellular Ca(2+). The steroidogenic effect of thapsigargin was inhibited by Zn(2+) and Cd(2+) but not by nifedipine. These results suggest that there is, in bovine adrenocortical fasciculata cells, a steroidogenesis-linked Ca(2+) entry process other than that involving voltage-operated Ca(2+) channels and that the process might be capacitative Ca(2+) entry.

scholarly journals Sestrin2-Mediated Autophagy Contributes to Drug Resistance via Endoplasmic Reticulum Stress in Human Osteosarcoma

2021 ◽  
Vol 9 ◽  
Author(s):  
Zhen Tang ◽  
Xinghui Wei ◽  
Tian Li ◽  
Wei Wang ◽  
Hao Wu ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Endoplasmic Reticulum Stress ◽  
Cell Apoptosis ◽  
Tumour Progression ◽  
Osteosarcoma Cell ◽  
Osteosarcoma Cells ◽  
Gene Target ◽  
Human Osteosarcoma ◽  
Human Osteosarcoma Cells

One contributor to the high mortality of osteosarcoma is its reduced sensitivity to chemotherapy, but the mechanism involved is unclear. Improving the sensitivity of osteosarcoma to chemotherapy is urgently needed to improve patient survival. We found that chemotherapy triggered apoptosis of human osteosarcoma cells in vitro and in vivo; this was accompanied by increased Sestrin2 expression. Importantly, autophagy was also enhanced with increased Sestrin2 expression. Based on this observation, we explored the potential role of Sestrin2 in autophagy of osteosarcoma. We found that Sestrin2 inhibited osteosarcoma cell apoptosis by promoting autophagy via inhibition of endoplasmic reticulum stress, and this process is closely related to the PERK-eIF2α-CHOP pathway. In addition, our study showed that low Sestrin2 expression can effectively reduce autophagy of human osteosarcoma cells after chemotherapy, increase p-mTOR expression, decrease Bcl-2 expression, promote osteosarcoma cell apoptosis, and slow down tumour progression in NU/NU mice. Sestrin2 activates autophagy by inhibiting mTOR via the PERK-eIF2α-CHOP pathway and inhibits apoptosis via Bcl-2. Therefore, our results explain one underlying mechanism of increasing the sensitivity of osteosarcoma to chemotherapy and suggest that Sestrin2 is a promising gene target.

scholarly journals In vitro effects of compounds isolated from Sideritis brevibracteata on bovine kidney cortex glutathione reductase.

2011 ◽  
Vol 58 (4) ◽  
Author(s):  
Berivan Tandogan ◽  
Ayşegül Güvenç ◽  
İhsan Çalış ◽  
Nuriye Nuray Ulusu
Keyword(s):  
Glutathione Reductase ◽  
Reduced Glutathione ◽  
Kidney Cortex ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Bovine Kidney ◽  
Aerial Parts ◽  
In Vitro Effects

Glutathione reductase (GR, E.C 1.6.4.2) is a flavoprotein that catalyzes NADPH-dependent reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH). The aim of this study was to investigate in vitro effects of phenolic compounds isolated from Sideritis brevibracteata on bovine kidney GR. The Sideritis species are widely found in nature and commonly used as medicinal plants. 7-O-glycosides of 8-OH-flavones (hypolaetin, isoscutellarein and 3'-hydroxy-4'-O-methylisoscutellarein) were isolated from aerial parts of Sideritis brevibracteata. These compounds inhibited bovine kidney cortex GR in a concentration-dependent manner. Kinetic characterization of the inhibition was also performed.

In vitro Effects of Chlorpyrifos on the Acetylcholinesterase Activity of Euryhaline Fish, Oreochromis mossambicus

2010 ◽  
Vol 65 (3-4) ◽  
pp. 303-306 ◽  
Author(s):  
Janapala Venkateswara Rao ◽  
Pathakoti Kavitha
Keyword(s):  
Ache Activity ◽  
Kinetic Studies ◽  
Gill Tissue ◽  
Acetylcholinesterase Activity ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
In Vitro Effects ◽  
Brain Ache Activity ◽  
Vitro Effect

The in vitro effect of a widely used organophosphorus insecticide, chlorpyrifos (CPP), on the acetylcholinesterase (AChE) activity was studied in vitro. The kinetic constants Km and Vmax and the bimolecular constant ki were determined in vitro. The in vitro AChE study indicated that CPP is neurotoxic and that it alters the apparent Km values widely in a concentration-dependent manner, resulting in a competitive type of inhibition. Based on the ki values, the sensitivity of AChE in brain is greater than that in gill tissue, at 7.3 · 10 - 5 M and 11.92 · 10 - 5 M, respectively. The study points to the importance of kinetic studies and the results suggest that in biomonitoring programmes brain AChE activity can be a good diagnostic tool for CPP toxicity.

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