Effects of cytotoxic agents on functional integrity and antioxidant enzymes in clonal beta-cells

SF Picton, Janie McCluskey, Peter Flatt, Neville McClenaghan

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Effects of cytotoxic agents and hydrogen peroxide were examined using pancreatic BRIN-BD11 cells and the parental insulinoma RINm5F cell line. Cell viability was determined using the MTT colorimetric assay and the TUNEL assay was used to assess apoptosis and acridine orange assay was used to determine levels of apoptosis versus necrosis. RT-PCR studies were employed to investigate the effects of the toxins on the expression of antioxidative enzymes, superoxide dismutase (SOD), glutathionine peroxidase (GPX) and catalase (CAT). Streptozotocin, hydrogen peroxide, alloxan and ninhydrin exerted time- and concentration-de pendent toxic effects on BRIN-BD11 and RINm5F cells. RT-PCR showed that 90 minutes exposure of BRIN-BD11 cells or RINm5F cells to 5 mM ninhydrin down regulates SOD, GPX and CAT antioxidative enzymes. Glutathionine peroxicase gene expression was also down regulated in both types of cell by hydrogen peroxide. There were no significant differences in antioxidant gene expression after exposure to the other toxins under the conditions employed. TUNEL assay revealed that streptozotocin (8 mM) and hydrogen peroxide (125 muM) had no significant effect on the number of cells undergoing apoptosis. However after exposure to ninhydrin (5 mM) almost 100% of the non-viable BRIN-BD11 cells and around 50% of the RINm5F cells were dying by apoptosis. With the BRIN-BD11 cells there was around a 30% increase in the number of apoptotic cells compared with 50% in the RINm5F cells after exposure to alloxan (16 mM). The results indicate multiple effects of cytotoxic agents on functional integrity and antioxidant enzyme gene expression in clonal beta-cells,
LanguageEnglish
PagesS70-S77
JournalDiabetes and Metabolism
Volume28
Issue number6, Par
Publication statusPublished - Dec 2002

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Cytotoxins
Antioxidants
Enzymes
Ninhydrin
Hydrogen Peroxide
Apoptosis
Alloxan
In Situ Nick-End Labeling
Streptozocin
Gene Expression
Catalase
Peroxidase
Superoxide Dismutase
Cell Count
Polymerase Chain Reaction
Acridine Orange
Insulinoma
Poisons
Cell Survival
Necrosis

Cite this

Picton, SF ; McCluskey, Janie ; Flatt, Peter ; McClenaghan, Neville. / Effects of cytotoxic agents on functional integrity and antioxidant enzymes in clonal beta-cells. In: Diabetes and Metabolism. 2002 ; Vol. 28, No. 6, Par. pp. S70-S77.
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abstract = "Effects of cytotoxic agents and hydrogen peroxide were examined using pancreatic BRIN-BD11 cells and the parental insulinoma RINm5F cell line. Cell viability was determined using the MTT colorimetric assay and the TUNEL assay was used to assess apoptosis and acridine orange assay was used to determine levels of apoptosis versus necrosis. RT-PCR studies were employed to investigate the effects of the toxins on the expression of antioxidative enzymes, superoxide dismutase (SOD), glutathionine peroxidase (GPX) and catalase (CAT). Streptozotocin, hydrogen peroxide, alloxan and ninhydrin exerted time- and concentration-de pendent toxic effects on BRIN-BD11 and RINm5F cells. RT-PCR showed that 90 minutes exposure of BRIN-BD11 cells or RINm5F cells to 5 mM ninhydrin down regulates SOD, GPX and CAT antioxidative enzymes. Glutathionine peroxicase gene expression was also down regulated in both types of cell by hydrogen peroxide. There were no significant differences in antioxidant gene expression after exposure to the other toxins under the conditions employed. TUNEL assay revealed that streptozotocin (8 mM) and hydrogen peroxide (125 muM) had no significant effect on the number of cells undergoing apoptosis. However after exposure to ninhydrin (5 mM) almost 100{\%} of the non-viable BRIN-BD11 cells and around 50{\%} of the RINm5F cells were dying by apoptosis. With the BRIN-BD11 cells there was around a 30{\%} increase in the number of apoptotic cells compared with 50{\%} in the RINm5F cells after exposure to alloxan (16 mM). The results indicate multiple effects of cytotoxic agents on functional integrity and antioxidant enzyme gene expression in clonal beta-cells,",
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Picton, SF, McCluskey, J, Flatt, P & McClenaghan, N 2002, 'Effects of cytotoxic agents on functional integrity and antioxidant enzymes in clonal beta-cells', Diabetes and Metabolism, vol. 28, no. 6, Par, pp. S70-S77.

Effects of cytotoxic agents on functional integrity and antioxidant enzymes in clonal beta-cells. / Picton, SF; McCluskey, Janie; Flatt, Peter; McClenaghan, Neville.

In: Diabetes and Metabolism, Vol. 28, No. 6, Par, 12.2002, p. S70-S77.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effects of cytotoxic agents on functional integrity and antioxidant enzymes in clonal beta-cells

AU - Picton, SF

AU - McCluskey, Janie

AU - Flatt, Peter

AU - McClenaghan, Neville

N1 - Workshop on the Endocrine Pancreas, BRUSSELS, BELGIUM, DEC 08, 2001

PY - 2002/12

Y1 - 2002/12

N2 - Effects of cytotoxic agents and hydrogen peroxide were examined using pancreatic BRIN-BD11 cells and the parental insulinoma RINm5F cell line. Cell viability was determined using the MTT colorimetric assay and the TUNEL assay was used to assess apoptosis and acridine orange assay was used to determine levels of apoptosis versus necrosis. RT-PCR studies were employed to investigate the effects of the toxins on the expression of antioxidative enzymes, superoxide dismutase (SOD), glutathionine peroxidase (GPX) and catalase (CAT). Streptozotocin, hydrogen peroxide, alloxan and ninhydrin exerted time- and concentration-de pendent toxic effects on BRIN-BD11 and RINm5F cells. RT-PCR showed that 90 minutes exposure of BRIN-BD11 cells or RINm5F cells to 5 mM ninhydrin down regulates SOD, GPX and CAT antioxidative enzymes. Glutathionine peroxicase gene expression was also down regulated in both types of cell by hydrogen peroxide. There were no significant differences in antioxidant gene expression after exposure to the other toxins under the conditions employed. TUNEL assay revealed that streptozotocin (8 mM) and hydrogen peroxide (125 muM) had no significant effect on the number of cells undergoing apoptosis. However after exposure to ninhydrin (5 mM) almost 100% of the non-viable BRIN-BD11 cells and around 50% of the RINm5F cells were dying by apoptosis. With the BRIN-BD11 cells there was around a 30% increase in the number of apoptotic cells compared with 50% in the RINm5F cells after exposure to alloxan (16 mM). The results indicate multiple effects of cytotoxic agents on functional integrity and antioxidant enzyme gene expression in clonal beta-cells,

AB - Effects of cytotoxic agents and hydrogen peroxide were examined using pancreatic BRIN-BD11 cells and the parental insulinoma RINm5F cell line. Cell viability was determined using the MTT colorimetric assay and the TUNEL assay was used to assess apoptosis and acridine orange assay was used to determine levels of apoptosis versus necrosis. RT-PCR studies were employed to investigate the effects of the toxins on the expression of antioxidative enzymes, superoxide dismutase (SOD), glutathionine peroxidase (GPX) and catalase (CAT). Streptozotocin, hydrogen peroxide, alloxan and ninhydrin exerted time- and concentration-de pendent toxic effects on BRIN-BD11 and RINm5F cells. RT-PCR showed that 90 minutes exposure of BRIN-BD11 cells or RINm5F cells to 5 mM ninhydrin down regulates SOD, GPX and CAT antioxidative enzymes. Glutathionine peroxicase gene expression was also down regulated in both types of cell by hydrogen peroxide. There were no significant differences in antioxidant gene expression after exposure to the other toxins under the conditions employed. TUNEL assay revealed that streptozotocin (8 mM) and hydrogen peroxide (125 muM) had no significant effect on the number of cells undergoing apoptosis. However after exposure to ninhydrin (5 mM) almost 100% of the non-viable BRIN-BD11 cells and around 50% of the RINm5F cells were dying by apoptosis. With the BRIN-BD11 cells there was around a 30% increase in the number of apoptotic cells compared with 50% in the RINm5F cells after exposure to alloxan (16 mM). The results indicate multiple effects of cytotoxic agents on functional integrity and antioxidant enzyme gene expression in clonal beta-cells,

M3 - Article

VL - 28

SP - S70-S77

JO - Diabetes and Metabolism

T2 - Diabetes and Metabolism

JF - Diabetes and Metabolism

SN - 1262-3636

IS - 6, Par

ER -