Separation of pigmented and albino melanocytes and the concomitant evaluation of endogenous peroxide content using flow cytometry

Boissy, Raymond E.; Trinkle, Linda S.; Nordlund, James J.

doi:10.1002/cyto.990100616

Cited by 37 publications

(20 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fifty M DCHF (Mobitech) dissolved in dimethylformamide were added and cells incubated for 15 min at 37°C. Thereafter, cells were placed on ice and aliquots of 10,000 cells scanned in a Becton Dickinson FACSSORT according to Boissy et al (1989) released from approximately 2 ϫ 10 5 HeLa or F26 cells grown in cuvettes was determined as described previously (McCord and Fridovich, 1969;Loschen et al, 1971;Meier et al, 1989Meier et al, , 1990Schreck et al, 1992a).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Induction of Oxidative Stress by Okadaic Acid Is Required for Activation of Transcription Factor NF-κB

Schmidt

Traenckner

Meier

et al. 1995

Journal of Biological Chemistry

168

101

View full text Add to dashboard Cite

The widely used phosphatase 1 and 2A inhibitor okadaic acid is one of the many stimuli activating transcription factor NF-B in cultured cells. Phosphorylation of IB-␣, one of NF-B's inhibitory subunits, is a prerequisite for IB degradation and the subsequent liberation of transcriptionally active NF-B. This observation suggested that the phosphorylation status of IB is influenced by an okadaic acid-sensitive phosphatase. In this study, we provide evidence that the effect of okadaic acid on NF-B activation is indirect and dependent on the production of reactive oxygen intermediates rather than the inhibition of an IB-␣ phosphatase. Okadaic acid was found to be a strong inducer of cellular H 2 O 2 and superoxide production in two distinct cell lines. The structurally unrelated phosphatase inhibitor calyculin A also induced oxidative stress. The delayed onset of reactive oxygen production in response to okadaic acid correlated with the delayed activation of NF-B. Moreover, NF-B induction was optimal at the same okadaic acid concentration that caused optimal H 2 O 2 production. Both reactive oxygen intermediates production and NF-B activation were inhibited by the antioxidant pyrrolidine dithiocarbamate and 8-(diethylamino)octyl-3,4,5-trimethyoxybenzoate, a Ca 2؉ chelator. Future experiments using phosphatase inhibitors in intact cells must consider that the compounds can act as strong inducers of oxidative stress, which provides one explanation for their tumor-promoting activity.

show abstract

Section: Methodsmentioning

confidence: 99%

“…OA Increases Cellular ROI Production which Is Prevented by PDTC-DCFH is a dye that allows to monitor cellular ROI production by FACS analysis (Boissy et al, 1989). As shown in Fig.…”

Section: Oa Activates a Prototypic Nf-b Complex By Ib-␣ Degradation-inmentioning

confidence: 99%

Induction of Oxidative Stress by Okadaic Acid Is Required for Activation of Transcription Factor NF-κB

Schmidt

Traenckner

Meier

et al. 1995

Journal of Biological Chemistry

168

101

View full text Add to dashboard Cite

show abstract

“…ROS Determination. ROS was measured by a dichlorofluorescein assay described by Boissy et al (1989). In brief, cells were grown to ϳ80% confluence and incubated with 2 M DCFH-DA.…”

Section: C-abl Is Required For Curcumin-induced Apoptosis 63mentioning

confidence: 99%

c-Abl Kinase Regulates Curcumin-Induced Cell Death through Activation of c-Jun N-Terminal Kinase

Kamath

Jiang²,

Sun³

et al. 2006

Mol Pharmacol

View full text Add to dashboard Cite

Curcumin, a natural phenolic compound found in turmeric (Curcuma longa) exhibits anticancer properties, attributed to its antiproliferative and apoptosis-inducing activity. The ubiquitously expressed nonreceptor tyrosine kinase c-Abl regulates stress responses induced by oxidative agents such as ionizing radiation and H 2 O 2 . In this study, we show that c-Abl is an important component of the cell death response activated by curcumin and that Abl mediates this response partly through activation of c-Jun N-terminal kinase (JNK). Therefore, inhibition of Abl by STI571 [imatinib (Gleevec)] treatment or downregulation of Abl expression through Abl-specific short-hairpin RNA (shRNA) diminished cell death induction and JNK activation. Highlighting the interdependent nature of the Abl and JNK signaling in the curcumin-induced cell death response, a JNK inhibitor [anthra(1,9-cd)pyrazol-6(2H)-one-1,9-pyrazoloanthrone (SP600125)] caused very little cell death inhibition in STI571-pretreated cells and in Abl shRNA-expressing cells. Moreover, treatment with Abl and JNK inhibitor alone or together caused similar levels of cell death inhibition. Although p53 induction in response to curcumin treatment is dependent on Abl, we found that Abl3p53 signaling is not necessary for curcumin-induced cell death. Taken together, the results demonstrate the differential roles played by Abl3p53 and Abl3 JNK signaling events in modulating the cell death response to curcumin.

show abstract

“…Cells (1 ϫ 10 6 /ml) were loaded with 10 M DCFH-DA, and analyzed by Becton Dickenson FACSCAN with excitation and emission settings of 495 and 525 nm, respectively (16).…”

Section: Fluorescent Measurement Of Intracellular Reactive Oxygen Spementioning

confidence: 99%

BCL-2 expression or antioxidants prevent hyperglycemia-induced formation of intracellular advanced glycation endproducts in bovine endothelial cells.

Giardino¹,

Edelstein²,

Brownlee³

1996

J. Clin. Invest.

221

121

View full text Add to dashboard Cite

Hyperglycemia rapidly induces an increase in intracellular advanced glycation end products (AGEs) in bovine endothelial cells, causing an alteration in bFGF activity (Giardino, I., D. Edelstein, and M. Brownlee. 1994. J. Clin. Invest. 94:110-117). Because sugar or sugar-adduct autoxidation is critical for AGE formation in vitro, we evaluated the role of reactive oxygen species (ROS) in intracellular AGE formation, using bovine aortic endothelial cells. 30 mM glucose increased intracellular ROS formation by 250% and lipid peroxidation by 330%, while not affecting ROS in the media. In cells depleted of glutathione, intracellular AGE accumulation increased linearly with ROS generation as measured by immunoblotting and the fluorescent probe DCFH (AGE 0.258-3.531 AU* mm ր 5 ϫ 10 4 cells, DCF 57-149 mean AU, r ϭ .998, P Ͻ .002). Deferoxamine, ␣ -tocopherol, and dimethylsulfoxide each inhibited hyperglycemia-induced formation of both ROS and AGE. To differentiate an effect of ROS generation on AGE formation from an effect of more distal oxidative processes, GM7373 endothelial cell lines were generated that stably expressed the peroxidationsuppressing proto-oncogene bcl-2. bcl-2 had no effect on hyperglycemia-induced intracellular ROS formation. In contrast, bcl-2 expression decreased both lipid peroxidation (100% at 3 h and 29% at 168 h) and AGE formation (55% at 168 h). These data show that a ROS-dependent process plays a central role in the generation of intracellular AGEs, and that inhibition of oxidant pathways prevents intracellular AGE formation. (

show abstract

Separation of pigmented and albino melanocytes and the concomitant evaluation of endogenous peroxide content using flow cytometry

Cited by 37 publications

References 22 publications

Induction of Oxidative Stress by Okadaic Acid Is Required for Activation of Transcription Factor NF-κB

Induction of Oxidative Stress by Okadaic Acid Is Required for Activation of Transcription Factor NF-κB

c-Abl Kinase Regulates Curcumin-Induced Cell Death through Activation of c-Jun N-Terminal Kinase

BCL-2 expression or antioxidants prevent hyperglycemia-induced formation of intracellular advanced glycation endproducts in bovine endothelial cells.

Contact Info

Product

Resources

About