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Article ; Online: Effects of cyclopiazonic acid and dexamethasone on serotonin-induced calcium responses in vascular smooth muscle cells.

Selli, Cigdem / Tosun, Metiner

2016 Volume 72, Issue 2, Page(s) 245–253

Abstract: We previously observed that sarcoendoplasmic reticulum Ca(2+) ATPase (SERCA) blockade by cyclopiazonic acid (CPA) significantly potentiates serotonin (5-hydroxytryptamine (5-HT))-induced vascular contractions. Furthermore, 5-HT receptor antagonist ... ...

Abstract	We previously observed that sarcoendoplasmic reticulum Ca(2+) ATPase (SERCA) blockade by cyclopiazonic acid (CPA) significantly potentiates serotonin (5-hydroxytryptamine (5-HT))-induced vascular contractions. Furthermore, 5-HT receptor antagonist methysergide partially inhibited CPA-potentiated 5-HT contractions. In the present study, we further investigated whether SERCA inhibition potentiates 5-HT-induced Ca(2+) responses along with attenuating the receptor antagonism by store-operated Ca(2+) (SOC) entry and protein kinase C (PKC)-mediated mechanisms. The effects of dexamethasone that was previously shown to induce SOC entry and enhance 5-HT responses were also tested. For this purpose, intracellular Ca(2+) levels were monitored in A7r5 embryonic rat vascular smooth muscle cells by spectrofluorometry using the fluorescent indicator fura-2. The results showed that CPA, although not dexamethasone, significantly potentiated 5-HT-induced Ca(2+) elevations. Ketanserin partially decreased 5-HT-induced and CPA-potentiated Ca(2+) elevations whereas both PKC inhibitor D-sphingosine and SOC entry blocker 2-aminoethoxydiphenyl borate (2-APB) abolished the remaining responses. The data suggests that diminished antagonistic effect on 5-HT-induced Ca(2+) elevations in the presence of SERCA inhibition is induced by SOC entry and PKC activation.
MeSH term(s)	Animals ; Anti-Inflammatory Agents/chemistry ; Anti-Inflammatory Agents/pharmacology ; Calcium Channel Blockers/pharmacology ; Calcium Signaling/drug effects ; Cell Line ; Dexamethasone/antagonists & inhibitors ; Dexamethasone/pharmacology ; Enzyme Inhibitors/chemistry ; Enzyme Inhibitors/pharmacology ; Indoles/antagonists & inhibitors ; Indoles/pharmacology ; Ketanserin/pharmacology ; Methysergide/pharmacology ; Muscle, Smooth, Vascular/drug effects ; Muscle, Smooth, Vascular/metabolism ; Protein Kinase C/antagonists & inhibitors ; Protein Kinase C/metabolism ; Protein Kinase Inhibitors/pharmacology ; Rats ; Sarcoplasmic Reticulum Calcium-Transporting ATPases/antagonists & inhibitors ; Sarcoplasmic Reticulum Calcium-Transporting ATPases/metabolism ; Serotonin/chemistry ; Serotonin/metabolism ; Serotonin Antagonists/pharmacology ; Serotonin Receptor Agonists/chemistry ; Serotonin Receptor Agonists/pharmacology ; Vasoconstrictor Agents/antagonists & inhibitors ; Vasoconstrictor Agents/pharmacology ; Vasodilator Agents/antagonists & inhibitors ; Vasodilator Agents/pharmacology
Chemical Substances	Anti-Inflammatory Agents ; Calcium Channel Blockers ; Enzyme Inhibitors ; Indoles ; Protein Kinase Inhibitors ; Serotonin Antagonists ; Serotonin Receptor Agonists ; Vasoconstrictor Agents ; Vasodilator Agents ; Serotonin (333DO1RDJY) ; Dexamethasone (7S5I7G3JQL) ; Ketanserin (97F9DE4CT4) ; Protein Kinase C (EC 2.7.11.13) ; Sarcoplasmic Reticulum Calcium-Transporting ATPases (EC 3.6.3.8) ; cyclopiazonic acid (X9TLY4580Z) ; Methysergide (XZA9HY6Z98)
Language	English
Publishing date	2016-06
Publishing country	Spain
Document type	Comparative Study ; Journal Article
ZDB-ID	1325104-1
ISSN	1877-8755 ; 0034-9402 ; 1138-7548
ISSN (online)	1877-8755
ISSN	0034-9402 ; 1138-7548
DOI	10.1007/s13105-016-0474-8
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Article: Varenicline Prevents LPS-Induced Inflammatory Response via Nicotinic Acetylcholine Receptors in RAW 264.7 Macrophages.

Baris, Elif / Efe, Hande / Gumustekin, Mukaddes / Arici, Mualla Aylin / Tosun, Metiner

Frontiers in molecular biosciences

2021 Volume 8, Page(s) 721533

Abstract: The cholinergic anti-inflammatory pathway plays an important role in controlling inflammation. This study investigated the effects of varenicline, an α7 nicotinic acetylcholine receptor (α7nAChR) agonist, on inflammatory cytokine levels, cell ... ...

Abstract	The cholinergic anti-inflammatory pathway plays an important role in controlling inflammation. This study investigated the effects of varenicline, an α7 nicotinic acetylcholine receptor (α7nAChR) agonist, on inflammatory cytokine levels, cell proliferation, and migration rates in a lipopolysaccharide (LPS)-induced inflammation model in RAW 264.7 murine macrophage cell lines. The cells were treated with increasing concentrations of varenicline, followed by LPS incubation for 24 h. Prior to receptor-mediated events, anti-inflammatory effects of varenicline on different cytokines and chemokines were investigated using a cytokine array. Nicotinic AChR-mediated effects of varenicline were investigated by using a non-selective nAChR antagonist mecamylamine hydrochloride and a selective α7nAChR antagonist methyllycaconitine citrate. TNFα, IL-1β, and IL-6 levels were determined by the ELISA test in cell media 24 h after LPS administration and compared with those of dexamethasone. The rates of cellular proliferation and migration were monitored for 24 h after drug treatment using a real-time cell analysis system. Varenicline decreased LPS-induced cytokines and chemokines including TNFα, IL-6, and IL-1β via α7nAChRs to a similar level that observed with dexamethasone. Varenicline treatment decreased LPS-induced cell proliferation, without any nAChR involvement. On the other hand, the LPS-induced cell migration rate decreased with varenicline via α7nAChR. Our data suggest that varenicline inhibits LPS-induced inflammatory response by activating α7nAChRs within the cholinergic anti-inflammatory pathway, reducing the cytokine levels and cell migration.
Language	English
Publishing date	2021-10-12
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2814330-9
ISSN	2296-889X
ISSN	2296-889X
DOI	10.3389/fmolb.2021.721533
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Article: Implications of Possible HBV-Driven Regulation of Gene Expression in Stem Cell-like Subpopulation of Huh-7 Hepatocellular Carcinoma Cell Line.

Demir, Ayse Banu / Benvenuto, Domenico / Karacicek, Bilge / Erac, Yasemin / Spoto, Silvia / Angeletti, Silvia / Ciccozzi, Massimo / Tosun, Metiner

Journal of personalized medicine

2022 Volume 12, Issue 12

Abstract: Elevated levels of STIM1, an endoplasmic reticulum ... ...

Abstract	Elevated levels of STIM1, an endoplasmic reticulum Ca
Language	English
Publishing date	2022-12-14
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662248-8
ISSN	2075-4426
ISSN	2075-4426
DOI	10.3390/jpm12122065
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Article: Effects of cell seeding density on real-time monitoring of anti-proliferative effects of transient gene silencing.

Selli, Cigdem / Erac, Yasemin / Tosun, Metiner

Journal of biological research (Thessalonike, Greece)

2016 Volume 23, Page(s) 20

Abstract: Background: Real-time cellular analysis systems enable impedance-based label-free and dynamic monitoring of various cellular events such as proliferation. In this study, we describe the effects of initial cell seeding density on the anti-proliferative ... ...

Abstract	Background: Real-time cellular analysis systems enable impedance-based label-free and dynamic monitoring of various cellular events such as proliferation. In this study, we describe the effects of initial cell seeding density on the anti-proliferative effects of transient gene silencing monitored via real-time cellular analysis. We monitored the real-time changes in proliferation of Huh7 hepatocellular carcinoma and A7r5 vascular smooth muscle cells with different initial seeding densities following transient receptor potential canonical 1 (TRPC1) silencing using xCELLigence system. Huh7 and A7r5 cells were seeded on E-plate 96 at 10,000, 5000, 1250 and 5000, 2500 cells well Results: TRPC1 silencing did not inhibit the proliferation rates of Huh7 cells at 10,000 cells well Conclusions: Data suggest that the effects of transient silencing on cell proliferation differ depending on the initial cell seeding density. While high seeding densities mask the significant changes in proliferation, the inhibitory effects of silencing become apparent at lower seeding densities as the entry into log phase is delayed. Using the optimal initial seeding density is crucial when studying the effects of transient gene silencing. In addition, the results suggest that TRPC1 may contribute to proliferation and phenotypic switching of vascular smooth muscle cells.
Language	English
Publishing date	2016-12-01
Publishing country	Greece
Document type	Journal Article
ZDB-ID	2186880-3
ISSN	2241-5793 ; 1790-045X
ISSN (online)	2241-5793
ISSN	1790-045X
DOI	10.1186/s40709-016-0057-4
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Article: Effects of 1-(2-trifluoromethylphenyl)-imidazole (TRIM) on receptor-independent and -dependent contractile responses in rat aorta.

Selli, Çiğdem / Eraç, Yasemin / Tosun, Metiner

Turkish journal of medical sciences

2016 Volume 46, Issue 4, Page(s) 1209–1214

Abstract: Background/aim: This study investigates whether 1-(2-trifluoromethylphenyl)-imidazole (TRIM), originally proposed as a nitric oxide synthase inhibitor and also suggested to be an inhibitor of store-operated calcium entry in mouse anococcygeal muscle, ... ...

Abstract	Background/aim: This study investigates whether 1-(2-trifluoromethylphenyl)-imidazole (TRIM), originally proposed as a nitric oxide synthase inhibitor and also suggested to be an inhibitor of store-operated calcium entry in mouse anococcygeal muscle, inhibits receptor-independent and -dependent responses in rat thoracic aorta. Materials and methods: Cyclopiazonic acid- and serotonin-induced vascular responses were investigated in aortic segments isolated from male Sprague Dawley rats using isolated tissue experiments. Changes in intracellular calcium levels were also monitored via front surface fluorescence measurements in fura-2-loaded embryonic rat vascular smooth muscle cell line A7r5. Results: TRIM inhibited serotonin-mediated vascular contractions without affecting cyclopiazonic acid-induced responses. In addition, TRIM caused a nonlinear rightward shift in the serotonin concentration-response curve, possibly via serotonin receptor modulation. Conclusion: TRIM may have an impact on investigation of tissue-specific receptor-independent and -dependent vascular responses. It may also be used as a lead compound in the development of selective serotonin receptor modulators.
MeSH term(s)	Animals ; Aorta ; Calcium ; Imidazoles ; Male ; Muscle Contraction ; Muscle, Smooth, Vascular ; Rats ; Rats, Sprague-Dawley
Chemical Substances	Imidazoles ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2016-06-23
Publishing country	Turkey
Document type	Journal Article
ZDB-ID	1183461-4
ISSN	1303-6165 ; 1300-0144
ISSN (online)	1303-6165
ISSN	1300-0144
DOI	10.3906/sag-1502-109
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Article ; Online: Simultaneous measurement of cytosolic and mitochondrial calcium levels: observations in TRPC1-silenced hepatocellular carcinoma cells.

Selli, Cigdem / Erac, Yasemin / Tosun, Metiner

Journal of pharmacological and toxicological methods

2015 Volume 72, Page(s) 29–34

Abstract: Introduction: The measurement of intracellular Ca(2+), cytosolic or stored in organelles, i.e., mitochondria, gave valuable data for numerous areas of research. In case of tumor cells, mitochondrial Ca(2+) levels play essential roles in apoptosis along ... ...

Abstract	Introduction: The measurement of intracellular Ca(2+), cytosolic or stored in organelles, i.e., mitochondria, gave valuable data for numerous areas of research. In case of tumor cells, mitochondrial Ca(2+) levels play essential roles in apoptosis along with endoplasmic reticulum (ER) Ca(2+). In this study, we describe a Ca(2+) monitoring system that allows studying both adherent cells and tissues and discuss data obtained from hepatocellular carcinoma cells and rat thoracic aorta by using this system. Methods: For this purpose, two apparatus, one for adherent cells and the other for intact rat aorta, were designed and produced. With this system, changes in cytosolic Ca(2+) levels following store-operated calcium (SOC) entry induced by sarco/endoplasmic reticulum Ca(2+) ATPase (SERCA) blockers were recorded in different hepatocellular carcinoma cells. Furthermore, cytosolic and mitochondrial Ca(2+) levels were simultaneously measured in TRPC1-silenced Huh7 hepatocellular carcinoma cells. In addition, the effects of trifluoromethylphenylimidazole (TRIM) on cyclopiazonic acid (CPA)-, serotonin (5-HT)-, and phenylephrine (PE)-induced changes in isometric force and cytosolic Ca(2+) levels were determined simultaneously in rat thoracic aorta. The effects of aging on PE-induced responses were also investigated. Results: After SOC entry activation, cytosolic Ca(2+) levels were increased, as expected in all hepatocellular carcinoma cells. Mitochondrial Ca(2+) levels following CPA-induced ER depletion were significantly (p<.05) diminished in TRPC1-silenced Huh7 cells. In addition, TRIM partially inhibited both 5-HT-induced contractions and cytosolic Ca(2+) levels without affecting CPA and PE responses. PE-induced contractions and cytosolic Ca(2+) levels were similar in aorta from young and old (3 and 22 months, respectively) rats. Discussion: We confirmed that the system provides valuable data about intracellular Ca(2+) dynamics by allowing simultaneous measurements and sequential addition of compounds in adherent cells. The decrease in mitochondrial Ca(2+) loading following CPA-induced ER depletion in TRPC1-silenced Huh7 cells suggests a possible role of TRPC1 in hepatocellular carcinoma cell apoptosis. The system also enables the simultaneous measurement of isometric force and cytosolic Ca(2+) levels and promotes understanding vascular physiology and disease.
MeSH term(s)	Animals ; Aorta/drug effects ; Aorta/physiology ; Calcium/chemistry ; Calcium/metabolism ; Carcinoma, Hepatocellular/metabolism ; Cell Line, Tumor ; Cytosol/chemistry ; Cytosol/metabolism ; Endoplasmic Reticulum/drug effects ; Endoplasmic Reticulum/physiology ; Gene Silencing ; Humans ; Indoles/pharmacology ; Liver Neoplasms/metabolism ; Mitochondria/chemistry ; Mitochondria/metabolism ; Phenylephrine/pharmacology ; Rats ; Serotonin/pharmacology ; TRPC Cation Channels/genetics ; TRPC Cation Channels/metabolism ; Vasoconstriction/drug effects ; Vasoconstriction/physiology
Chemical Substances	Indoles ; TRPC Cation Channels ; transient receptor potential cation channel, subfamily C, member 1 ; Phenylephrine (1WS297W6MV) ; Serotonin (333DO1RDJY) ; Calcium (SY7Q814VUP) ; cyclopiazonic acid (X9TLY4580Z)
Language	English
Publishing date	2015-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1105919-9
ISSN	1873-488X ; 1056-8719
ISSN (online)	1873-488X
ISSN	1056-8719
DOI	10.1016/j.vascn.2014.12.004
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Article: Simultaneous measurement of cytosolic and mitochondrial calcium levels: Observations in TRPC1-silenced hepatocellular carcinoma cells

Selli, Cigdem / Erac, Yasemin / Tosun, Metiner

Journal of pharmacological and toxicological methods. 2015 Mar., Apr., v. 72

2015

Abstract: The measurement of intracellular Ca²⁺, cytosolic or stored in organelles, i.e., mitochondria, gave valuable data for numerous areas of research. In case of tumor cells, mitochondrial Ca²⁺ levels play essential roles in apoptosis along with endoplasmic ... ...

Abstract	The measurement of intracellular Ca²⁺, cytosolic or stored in organelles, i.e., mitochondria, gave valuable data for numerous areas of research. In case of tumor cells, mitochondrial Ca²⁺ levels play essential roles in apoptosis along with endoplasmic reticulum (ER) Ca²⁺. In this study, we describe a Ca²⁺ monitoring system that allows studying both adherent cells and tissues and discuss data obtained from hepatocellular carcinoma cells and rat thoracic aorta by using this system.For this purpose, two apparatus, one for adherent cells and the other for intact rat aorta, were designed and produced. With this system, changes in cytosolic Ca²⁺ levels following store-operated calcium (SOC) entry induced by sarco/endoplasmic reticulum Ca²⁺ ATPase (SERCA) blockers were recorded in different hepatocellular carcinoma cells. Furthermore, cytosolic and mitochondrial Ca²⁺ levels were simultaneously measured in TRPC1-silenced Huh7 hepatocellular carcinoma cells. In addition, the effects of trifluoromethylphenylimidazole (TRIM) on cyclopiazonic acid (CPA)-, serotonin (5-HT)-, and phenylephrine (PE)-induced changes in isometric force and cytosolic Ca²⁺ levels were determined simultaneously in rat thoracic aorta. The effects of aging on PE-induced responses were also investigated.After SOC entry activation, cytosolic Ca²⁺ levels were increased, as expected in all hepatocellular carcinoma cells. Mitochondrial Ca²⁺ levels following CPA-induced ER depletion were significantly (p<.05) diminished in TRPC1-silenced Huh7 cells. In addition, TRIM partially inhibited both 5-HT-induced contractions and cytosolic Ca²⁺ levels without affecting CPA and PE responses. PE-induced contractions and cytosolic Ca²⁺ levels were similar in aorta from young and old (3 and 22months, respectively) rats.We confirmed that the system provides valuable data about intracellular Ca²⁺ dynamics by allowing simultaneous measurements and sequential addition of compounds in adherent cells. The decrease in mitochondrial Ca²⁺ loading following CPA-induced ER depletion in TRPC1-silenced Huh7 cells suggests a possible role of TRPC1 in hepatocellular carcinoma cell apoptosis. The system also enables the simultaneous measurement of isometric force and cytosolic Ca²⁺ levels and promotes understanding vascular physiology and disease.
Keywords	adenosinetriphosphatase ; aorta ; apoptosis ; calcium ; cyclopiazonic acid ; endoplasmic reticulum ; hepatoma ; mitochondria ; neoplasm cells ; phenylephrine ; physiology ; rats ; serotonin ; toxicology
Language	English
Dates of publication	2015-03
Size	p. 29-34.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	1105919-9
ISSN	1873-488X ; 1056-8719
ISSN (online)	1873-488X
ISSN	1056-8719
DOI	10.1016/j.vascn.2014.12.004
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Article ; Online: Differential expression of store-operated calcium- and proliferation-related genes in hepatocellular carcinoma cells following TRPC1 ion channel silencing.

Selli, Cigdem / Pearce, Dominic A / Sims, Andrew H / Tosun, Metiner

Molecular and cellular biochemistry

2016 Volume 420, Issue 1-2, Page(s) 129–140

Abstract: TRPC1 and store-operated Ca(2+) (SOC) entry have previously been associated with hepatocellular carcinoma cell proliferation. The aim of the study was to determine genes and processes associated with TRPC1 down-regulation and the resulting increase of ... ...

Abstract	TRPC1 and store-operated Ca(2+) (SOC) entry have previously been associated with hepatocellular carcinoma cell proliferation. The aim of the study was to determine genes and processes associated with TRPC1 down-regulation and the resulting increase of SOC entry and decrease in hepatocellular carcinoma cell proliferation. For this purpose, transcriptome analysis was performed to determine differentially expressed genes in TRPC1-silenced Huh7 cells. SOC entry- and proliferation-related genes correlated with TRPC1 down-regulation were also examined. Changes in SOC entry and cell proliferation were monitored in the TRPC1-silenced and parental cells and found to be significantly increased and decreased, respectively, in TRPC1-silenced cells. A total of 71 genes were significantly differentially expressed (40 up- and 31 down-regulated), including four mitogen-activated protein kinase (MAPK) signalling-associated genes. STIM1 levels were significantly up-regulated and negatively correlated with TRPC1 levels. In addition, expression of two cell cycle regulation genes, CDK11A/11B and URGCP, was observed to decrease, whereas ERBB3 and FGFR4, pro-survival genes, increased significantly in TRPC1-silenced cells. In conclusion, these results suggest reciprocal alterations in TRPC1 and STIM1 levels and a role for STIM1 in the regulation of SOC entry in TRPC1-silenced Huh7 cells. In addition to TRPC1, STIM1 may participate in Huh7 cell proliferation by regulating SOC entry. Alterations in MAPK signalling genes may be involved in diminished cell proliferation in TRPC1-silenced Huh7 cells. Similarly, changes in cell cycle regulating genes in TRPC1-silenced cells indicate possible cell cycle arrest along with compensatory up-regulation of ERBB3 growth factor receptor-amongst others-to maintain hepatocellular carcinoma cell proliferation.
MeSH term(s)	Carcinoma, Hepatocellular/genetics ; Carcinoma, Hepatocellular/metabolism ; Cell Cycle Checkpoints ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Humans ; Liver Neoplasms/genetics ; Liver Neoplasms/metabolism ; Neoplasm Proteins/biosynthesis ; Neoplasm Proteins/genetics ; TRPC Cation Channels/genetics ; TRPC Cation Channels/metabolism
Chemical Substances	Neoplasm Proteins ; TRPC Cation Channels ; transient receptor potential cation channel, subfamily C, member 1
Language	English
Publishing date	2016-09
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	184833-1
ISSN	1573-4919 ; 0300-8177
ISSN (online)	1573-4919
ISSN	0300-8177
DOI	10.1007/s11010-016-2776-0
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Article ; Online: Effects of passage number on proliferation and store-operated calcium entry in A7r5 vascular smooth muscle cells.

Erac, Yasemin / Selli, Cigdem / Filik, Pelin / Tosun, Metiner

Journal of pharmacological and toxicological methods

2014 Volume 70, Issue 1, Page(s) 1–5

Abstract: Introduction: Embryonic rat aortic smooth muscle cells, A7r5, have been used extensively as an in vitro vascular smooth muscle cell model. They are usually provided at 11th passage by supplier and generally used before 25th passage. However, the exact ... ...

Abstract	Introduction: Embryonic rat aortic smooth muscle cells, A7r5, have been used extensively as an in vitro vascular smooth muscle cell model. They are usually provided at 11th passage by supplier and generally used before 25th passage. However, the exact passage number (P#) used is not reported in general. Methods: In this study, A7r5 cells (P#<18 and P#>23) were used in quantitative gene (5-HT2A and SERCA2b) expression as well as in functional analyses including measurements of real-time changes in cell proliferation and in 5-HT- and CPA-induced intracellular Ca(2+) levels. Results: CPA-induced SR Ca(2+) release and store-operated Ca(2+) entry significantly increased (p<.05) while cell proliferation decreased in P#>23 cells (p<.01). Furthermore, 5-HT-induced Ca(2+) elevations and 5-HT2A mRNA levels did not change whereas SERCA2b mRNA levels and CPA-induced [Ca(2+)]i levels were significantly elevated in P#>23 cells (p<.05, p<.01, respectively). Discussion: Changes in SERCA2b expression and SOCE may contribute to suppression of cell proliferation during A7r5 subculturing. Therefore, passage numbers and subculturing procedure should be reported and taken into account during expressional and functional analyses for an accurate comparison of published data.
MeSH term(s)	Animals ; Aorta, Thoracic/metabolism ; Aorta, Thoracic/physiology ; Calcium/metabolism ; Cell Line ; Cell Proliferation/physiology ; Muscle, Smooth, Vascular/metabolism ; Muscle, Smooth, Vascular/physiology ; Myocytes, Smooth Muscle ; RNA, Messenger/metabolism ; Rats ; Sarcoplasmic Reticulum Calcium-Transporting ATPases/metabolism ; Serotonin/metabolism
Chemical Substances	Atp2a2 protein, rat ; RNA, Messenger ; Serotonin (333DO1RDJY) ; Sarcoplasmic Reticulum Calcium-Transporting ATPases (EC 3.6.3.8) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2014-03-12
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1105919-9
ISSN	1873-488X ; 1056-8719
ISSN (online)	1873-488X
ISSN	1056-8719
DOI	10.1016/j.vascn.2014.03.001
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Article: Effects of passage number on proliferation and store-operated calcium entry in A7r5 vascular smooth muscle cells

Erac, Yasemin / Selli, Cigdem / Filik, Pelin / Tosun, Metiner

Journal of pharmacological and toxicological methods. 2014 July, Aug., v. 70, no. 1

2014

Abstract: Embryonic rat aortic smooth muscle cells, A7r5, have been used extensively as an in vitro vascular smooth muscle cell model. They are usually provided at 11th passage by supplier and generally used before 25th passage. However, the exact passage number ( ... ...

Abstract	Embryonic rat aortic smooth muscle cells, A7r5, have been used extensively as an in vitro vascular smooth muscle cell model. They are usually provided at 11th passage by supplier and generally used before 25th passage. However, the exact passage number (P#) used is not reported in general.In this study, A7r5 cells (P#<18 and P#>23) were used in quantitative gene (5-HT₂A and SERCA2b) expression as well as in functional analyses including measurements of real-time changes in cell proliferation and in 5-HT- and CPA-induced intracellular Ca²⁺ levels.CPA-induced SR Ca²⁺ release and store-operated Ca²⁺ entry significantly increased (p<.05) while cell proliferation decreased in P#>23 cells (p<.01). Furthermore, 5-HT-induced Ca²⁺ elevations and 5-HT₂A mRNA levels did not change whereas SERCA2b mRNA levels and CPA-induced [Ca²⁺]ᵢ levels were significantly elevated in P#>23 cells (p<.05, p<.01, respectively).Changes in SERCA2b expression and SOCE may contribute to suppression of cell proliferation during A7r5 subculturing. Therefore, passage numbers and subculturing procedure should be reported and taken into account during expressional and functional analyses for an accurate comparison of published data.
Keywords	calcium ; cell proliferation ; genes ; models ; rats ; smooth muscle ; toxicology
Language	English
Dates of publication	2014-07
Size	p. 1-5.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	1105919-9
ISSN	1873-488X ; 1056-8719
ISSN (online)	1873-488X
ISSN	1056-8719
DOI	10.1016/j.vascn.2014.03.001
Database	NAL-Catalogue (AGRICOLA)

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