Glutathione S-transferase <i>π</i> in an arsenic-resistant Chinese hamster ovary cell line

Lo, Jeng-Fan; Wang, Huei-Feng; Tam, Ming F.; Lee, Te‐Chang

doi:10.1042/bj2880977

Cited by 54 publications

(46 citation statements)

References 43 publications

(36 reference statements)

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“…Furthermore, glutathione S-transferase π has been implicated in arsenic detoxification (Lo et al, 1992). We tested the impact of GST inhibition with ethacrynic acid on SK-Mel-28 cells.…”

Section: Resultsmentioning

confidence: 99%

Sensitivity to sodium arsenite in human melanoma cells depends upon susceptibility to arsenite-induced mitotic arrest

McNeely

Belshoff

Taylor

et al. 2008

Toxicology and Applied Pharmacology

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Arsenic induces clinical remission in patients with acute promyelocytic leukemia and has potential for treatment of other cancers. The current study examines factors influencing sensitivity to arsenic using human malignant melanoma cell lines. A375 and SK-Mel-2 cells were sensitive to clinically achievable concentrations of arsenite, whereas SK-Mel-3 and SK-Mel-28 cells required supratherapeutic levels for toxicity. Inhibition of glutathione synthesis, glutathione S-transferase (GST) activity, and multidrug resistance protein (MRP) transporter function attenuated arsenite resistance, consistent with studies suggesting arsenite is extruded from the cell as a glutathione conjugate by MRP-1. However, MRP-1 was not overexpressed in resistant lines and GST-π was only slightly elevated. ICP-MS analysis indicated arsenite-resistant SK-Mel-28 cells did not accumulate less arsenic than arsenite-sensitive A375 cells, suggesting resistance was not attributable to reduced arsenic accumulation but rather to intrinsic properties of resistant cell lines. The mode of arseniteinduced cell death was apoptosis. Arsenite-induced apoptosis is associated with cell cycle alterations. Cell cycle analysis revealed arsenite-sensitive cells arrested in mitosis whereas arsenite-resistant cells did not, suggesting induction of mitotic arrest occurs at lower intracellular arsenic concentrations. Higher intracellular arsenic levels induced cell cycle arrest in S-phase and G 2 -phase in SK-Mel-3 and SK-Mel-28 cells, respectively. The lack of arsenite-induced mitotic arrest in resistant cell lines was associated with a weakened spindle checkpoint resulting from reduced expression of spindle checkpoint protein BUBR1. These data suggest arsenite has potential for treatment of solid tumors but a functional spindle checkpoint is a prerequisite for a positive response to its clinical application.

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“…Furthermore, glutathione S-transferase π has been implicated in arsenic detoxification (Lo et al, 1992). We tested the impact of GST inhibition with ethacrynic acid on SK-Mel-28 cells.…”

Section: Resultsmentioning

confidence: 99%

Sensitivity to sodium arsenite in human melanoma cells depends upon susceptibility to arsenite-induced mitotic arrest

McNeely

Belshoff

Taylor

et al. 2008

Toxicology and Applied Pharmacology

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“…Our finding that GSTP1-1 was required for transport of As III (plus GSH) provides substantive evidence that this is likely to be the case. Overexpression of GSTP1-1 has been noted in several cell lines chronically exposed to As III , and it has been suggested that this overexpression facilitated the efflux of arsenic (24,48,49). However, the importance of GSTP1-1 in the efflux of arsenic from intact cells has been difficult to elucidate primarily because many of the compounds used to inhibit GSTP1-1 also inhibit MRP1 (e.g.…”

Section: Discussionmentioning

confidence: 99%

Arsenic Transport by the Human Multidrug Resistance Protein 1 (MRP1/ABCC1)

Leslie

Haimeur

Waalkes

2004

Journal of Biological Chemistry

244

197

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Inorganic arsenic is an established human carcinogen, but its metabolism is incompletely defined. The ATP binding cassette protein, multidrug resistance protein (MRP1/ABCC1), transports conjugated organic anions (e.g. leukotriene C 4 ) and also co-transports certain unmodified xenobiotics (e.g. vincristine) with glutathione (GSH). MRP1 also confers resistance to arsenic in association with GSH; however, the mechanism and the species of arsenic transported are unknown. Using membrane vesicles prepared from the MRP1-overexpressing lung cancer cell line, H69AR, we found that MRP1 transports arsenite (As III ) only in the presence of GSH but does not transport arsenate (As V ) (with or without GSH). The non-reducing GSH analogs L-␥-glutamyl-L-␣-aminobutyryl glycine and S-methyl GSH did not support As III transport, indicating that the free thiol group of GSH is required. GSH-dependent transport of As III was 2-fold higher at pH 6.5-7 than at a more basic pH, consistent with the formation and transport of the acidstable arsenic triglutathione (As(GS) 3 ). Immunoblot analysis of H69AR vesicles revealed the unexpected membrane association of GSH S-transferase P1-1 (GSTP1-1). Membrane vesicles from an MRP1-transfected HeLa cell line lacking membrane-associated GSTP1-1 did not transport As III even in the presence of GSH but did transport synthetic As(GS) 3 . The addition of exogenous GSTP1-1 to HeLa-MRP1 vesicles resulted in GSH-dependent As III transport. The apparent K m of As(GS) 3 for MRP1 was 0.32 M, suggesting a remarkably high relative affinity. As(GS) 3 transport by MRP1 was osmotically sensitive and was inhibited by several conjugated organic anions (MRP1 substrates) as well as the metalloid antimonite (K i 2.8 M). As(GS) 3 transport experiments using MRP1 mutants with substrate specificities differing from wild-type MRP1 suggested a commonality in the substrate binding pockets of As(GS) 3 and leukotriene C 4 . Finally, human MRP2 also transported As(GS) 3 . In conclusion, MRP1 transports inorganic arsenic as a tri-GSH conjugate, and GSTP1-1 may have a synergistic role in this process.

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“…Since arsenic is exported from the cell in an ATPdependent manner by the multidrug-resistance-associated protein (MRP), high expression of this membrane pump could also cause metal resistance. 43 Each of these proteins has been shown to be inducible by arsenic, or overexpressed in some arsenicresistant cell systems, 16,45,46 and their relative abundance may exert a combined effect on the response to As 2 O 3 . While neither GSTp, MRP, nor GPx were found to be overexpressed in NB4-M-AsR2 and NB4-M-AsR3 cells (data not shown), both resistant cell lines, and all cancer cells tested, regardless of GSH content or their initial response to As 2 O 3 , could be sensitized by intracellular GSH depletion.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, some reports suggest that modulation of GSH, and GSH utilization pathways, including upregulation of glutathione-Stransferase p (GSTp) and the multidrug resistance-associated protein (MRP), can confer metal resistance. [13][14][15][16][17][18][19][20] Owing to its known role as an environmental carcinogen, the question of how arsenic exerts its toxic effects has been the subject of intense study in recent years. In addition to causing mitochondrial toxicity, 21 impairing microtubule polymerization, 22 and deregulating a number of proteins and enzymes through sulfhydryl binding, 23 considerable evidence suggests that arsenic induces oxidative stress and the generation of hydrogen peroxide (H 2 O 2 ) and other reactive oxygen species (ROS).…”

Section: Introductionmentioning

confidence: 99%

Glutathione depletion overcomes resistance to arsenic trioxide in arsenic-resistant cell lines

et al. 2003

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Arsenic trioxide (As 2 O 3 ) is an effective treatment for acute promyelocytic leukemia (APL), but is less effective against other leukemias. Although the response of APL cells to As 2 O 3 has been linked to degradation of the PML/RARa fusion oncoprotein, there is evidence that PML/RARa expression is not the only mediator of arsenic sensitivity. Indeed, we found that exogenous expression of PML/RARa did not sensitize a non-APL leukemic line to As 2 O 3 . To evaluate possible other determinants of sensitivity of leukemic cells to As 2 O 3 , we derived two arsenic-resistant NB4 subclones. Despite being approximately 10-fold more resistant to arsenic than their parental cell line, PML/RARa protein was still degraded by As 2 O 3 in these cells, providing further evidence that loss of expression of the oncoprotein does not confer arsenic sensitivity. Both arsenic-resistant clones contained high glutathione (GSH) levels, however, and we found that GSH depletion coupled with As 2 O 3 treatment dramatically inhibited their growth. Annexin V-staining and TUNEL analysis confirmed a synergistic induction of apoptosis. In addition, these cells failed to accumulate ROS in response to arsenic treatment, in contrast to their arsenic-sensitive parental cells, unless cotreated with buthionine sulfoximine. While other malignant cells did not show a good correlation between arsenic sensitivity and GSH content, GSH depletion nevertheless sensitized all cell lines examined, regardless of their initial response to arsenic alone. These findings suggest that PML/RARa expression is not a determinant of arsenic sensitivity, and further support the coupling of GSH depletion and arsenic treatment as a novel treatment for human malignancies that are unresponsive to arsenic alone. Leukemia (2003) 17, 931-940.

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Glutathione S-transferase π in an arsenic-resistant Chinese hamster ovary cell line

Cited by 54 publications

References 43 publications

Sensitivity to sodium arsenite in human melanoma cells depends upon susceptibility to arsenite-induced mitotic arrest

Sensitivity to sodium arsenite in human melanoma cells depends upon susceptibility to arsenite-induced mitotic arrest

Arsenic Transport by the Human Multidrug Resistance Protein 1 (MRP1/ABCC1)

Glutathione depletion overcomes resistance to arsenic trioxide in arsenic-resistant cell lines

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