Functional evaluation of a fluorescent schweinfurthin: Mechanism of cytotoxicity and intracellular quantification

Craig H. Kuder, Ryan M. Sheehy, Jeffrey Neighbors, David F. Wiemer, Raymond Hohl

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Schweinfurthins are potent inhibitors of cancer cell growth, especially against human central nervous system tumor lines such as SF-295 cells. However, the mechanisms through which these compounds impede cell growth are not fully understood. In an effort to understand the basis for the effects of schweinfurthins, we present a fluorescent schweinfurthin, 3-deoxyschweinfurthin B-like p-nitro-bis-stilbene (3dSB-PNBS), which displays biological activity similar to that of 3-deoxyschweinfurthin B (3dSB). These two schweinfurthins retain the unique differential activity of the natural schweinfurthins, as evidenced by the spindle-like morphological changes induced in SF-295 cells and the unaltered appearance of human lung carcinoma A549 cells. We demonstrate that incubation with 3dSB or 3dSB-PNBS results in cleavage of poly-ADP-ribose polymerase (PARP) and caspase-9, both markers of apoptosis. Coincubation of 3dSB or 3dSB-PNBS with the caspase-9 inhibitor (Z)-Leu-Glu(O-methyl)-His-Asp(O- methyl)-fluoromethylketone prevents PARP cleavage. Therapeutic agents that induce apoptosis often activate cellular stress pathways. A marker for multiple stress pathways is the phosphorylation of eukaryotic initiation factor 2α, which is phosphorylated in response to 3dSB and 3dSB-PNBS treatment. Glucose-regulated protein 78 and protein disulfide isomerase, both endoplasmic reticulum chaperones, are up-regulated with schweinfurthin exposure. Using the fluorescent properties of 3dSB-PNBS and dimethoxyphenyl-p-nitro-bis-stilbene (DMP-PNBS), a control compound, we show that the intracellular levels of 3dSB-PNBS are higher than those of Rhodamine 123 or DMP-PNBS in SF-295 and A549 cells.

Original languageEnglish (US)
Pages (from-to)9-16
Number of pages8
JournalMolecular Pharmacology
Volume82
Issue number1
DOIs
StatePublished - Jul 1 2012

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Stilbenes
Poly(ADP-ribose) Polymerases
Caspase 9
Eukaryotic Initiation Factor-2
Apoptosis
Protein Disulfide-Isomerases
Rhodamine 123
Central Nervous System Neoplasms
Growth Inhibitors
Caspase Inhibitors
Endoplasmic Reticulum
3-deoxyschweinfurthin B-p-nitro bis-stilbene
Phosphorylation
Carcinoma
Lung
3-deoxyschweinfurthin B
Growth
Neoplasms
A549 Cells
Therapeutics

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Molecular Medicine

Cite this

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title = "Functional evaluation of a fluorescent schweinfurthin: Mechanism of cytotoxicity and intracellular quantification",
abstract = "Schweinfurthins are potent inhibitors of cancer cell growth, especially against human central nervous system tumor lines such as SF-295 cells. However, the mechanisms through which these compounds impede cell growth are not fully understood. In an effort to understand the basis for the effects of schweinfurthins, we present a fluorescent schweinfurthin, 3-deoxyschweinfurthin B-like p-nitro-bis-stilbene (3dSB-PNBS), which displays biological activity similar to that of 3-deoxyschweinfurthin B (3dSB). These two schweinfurthins retain the unique differential activity of the natural schweinfurthins, as evidenced by the spindle-like morphological changes induced in SF-295 cells and the unaltered appearance of human lung carcinoma A549 cells. We demonstrate that incubation with 3dSB or 3dSB-PNBS results in cleavage of poly-ADP-ribose polymerase (PARP) and caspase-9, both markers of apoptosis. Coincubation of 3dSB or 3dSB-PNBS with the caspase-9 inhibitor (Z)-Leu-Glu(O-methyl)-His-Asp(O- methyl)-fluoromethylketone prevents PARP cleavage. Therapeutic agents that induce apoptosis often activate cellular stress pathways. A marker for multiple stress pathways is the phosphorylation of eukaryotic initiation factor 2α, which is phosphorylated in response to 3dSB and 3dSB-PNBS treatment. Glucose-regulated protein 78 and protein disulfide isomerase, both endoplasmic reticulum chaperones, are up-regulated with schweinfurthin exposure. Using the fluorescent properties of 3dSB-PNBS and dimethoxyphenyl-p-nitro-bis-stilbene (DMP-PNBS), a control compound, we show that the intracellular levels of 3dSB-PNBS are higher than those of Rhodamine 123 or DMP-PNBS in SF-295 and A549 cells.",
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Functional evaluation of a fluorescent schweinfurthin : Mechanism of cytotoxicity and intracellular quantification. / Kuder, Craig H.; Sheehy, Ryan M.; Neighbors, Jeffrey; Wiemer, David F.; Hohl, Raymond.

In: Molecular Pharmacology, Vol. 82, No. 1, 01.07.2012, p. 9-16.

Research output: Contribution to journalArticle

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AU - Kuder, Craig H.

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