A splice variant of the human ion channel TRPM2 modulates neuroblastoma tumor growth through hypoxia-inducible factor (HIF)-1/2α

Shu Jen Chen, Nicholas E. Hoffman, Santhanam Shanmughapriy, Lei Bao, Kerry Keefer, Kathleen Conrad, Salim Merali, Yoshinori Takahashi, Thomas Abraham, Iwona Hirschler-Laszkiewicz, Ju Fang Wang, Xue Qian Zhang, Jianliang Song, Carlos Barrero, Yuguang Shi, Yuka Imamura Kawasawa, Michael Bayerl, Tianyu Sun, Mustafa Barbour, Hong Gang WangMuniswamy Madesh, Joseph Y. Cheung, Barbara A. Miller

Research output: Contribution to journalArticle

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Abstract

The calcium-permeable ion channel TRPM2 is highly expressed in a number of cancers. In neuroblastoma, full-length TRPM2 (TRPM2-L) protected cells from moderate oxidative stress through increased levels of forkhead box transcription factor 3a (FOXO3a) and superoxide dismutase 2. Cells expressing the dominant negative short isoform (TRPM2-S) had reduced FOXO3a and superoxide dismutase 2 levels, reduced calcium influx in response to oxidative stress, and enhanced reactive oxygen species, leading to decreased cell viability. Here, in xenografts generated with SH-SY5Y neuroblastoma cells stably expressing TRPM2 isoforms, growth of tumors expressing TRPM2-S was significantly reduced compared with tumors expressing TRPM2-L. Expression of hypoxia-inducible factor (HIF)-1/2α was significantly reduced in TRPM2-S-expressing tumor cells as was expression of target proteins regulated by HIF-1/2α including those involved in glycolysis (lactate dehydrogenaseAand enolase 2), oxidant stress (FOXO3a), angiogenesis (VEGF), mitophagy and mitochondrial function (BNIP3 and NDUFA4L2), and mitochondrial electron transport chain activity (cytochrome oxidase 4.1/4.2 in complex IV). The reduction in HIF-1/2α was mediated through both significantly reduced HIF-1/2α mRNA levels and increased levels of von Hippel-Lindau E3 ligase in TRPM2-S-expressing cells. Inhibition of TRPM2-L by pretreatment with clotrimazole or expression of TRPM2-S significantly increased sensitivity of cells to doxorubicin. Reduced survival of TRPM2-S-expressing cells after doxorubicin treatment was rescued by gain of HIF-1 or -2α function. These data suggest that TRPM2 activity is important for tumor growth and for cell viability and survival following doxorubicin treatment and that interference with TRPM2-L function may be a novel approach to reduce tumor growth through modulation of HIF-1/2α, mitochondrial function, and mitophagy.

Original languageEnglish (US)
Pages (from-to)36284-36302
Number of pages19
JournalJournal of Biological Chemistry
Volume289
Issue number52
DOIs
StatePublished - Jan 1 2014

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Hypoxia-Inducible Factor 1
Neuroblastoma
Ion Channels
Tumors
Growth
Cells
Doxorubicin
Mitochondrial Degradation
Neoplasms
Oxidative stress
Cell Survival
Protein Isoforms
Oxidative Stress
Calcium
Clotrimazole
Forkhead Transcription Factors
Ubiquitin-Protein Ligases
Phosphopyruvate Hydratase
Electron Transport Complex IV
Heterografts

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Chen, Shu Jen ; Hoffman, Nicholas E. ; Shanmughapriy, Santhanam ; Bao, Lei ; Keefer, Kerry ; Conrad, Kathleen ; Merali, Salim ; Takahashi, Yoshinori ; Abraham, Thomas ; Hirschler-Laszkiewicz, Iwona ; Wang, Ju Fang ; Zhang, Xue Qian ; Song, Jianliang ; Barrero, Carlos ; Shi, Yuguang ; Kawasawa, Yuka Imamura ; Bayerl, Michael ; Sun, Tianyu ; Barbour, Mustafa ; Wang, Hong Gang ; Madesh, Muniswamy ; Cheung, Joseph Y. ; Miller, Barbara A. / A splice variant of the human ion channel TRPM2 modulates neuroblastoma tumor growth through hypoxia-inducible factor (HIF)-1/2α. In: Journal of Biological Chemistry. 2014 ; Vol. 289, No. 52. pp. 36284-36302.
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title = "A splice variant of the human ion channel TRPM2 modulates neuroblastoma tumor growth through hypoxia-inducible factor (HIF)-1/2α",
abstract = "The calcium-permeable ion channel TRPM2 is highly expressed in a number of cancers. In neuroblastoma, full-length TRPM2 (TRPM2-L) protected cells from moderate oxidative stress through increased levels of forkhead box transcription factor 3a (FOXO3a) and superoxide dismutase 2. Cells expressing the dominant negative short isoform (TRPM2-S) had reduced FOXO3a and superoxide dismutase 2 levels, reduced calcium influx in response to oxidative stress, and enhanced reactive oxygen species, leading to decreased cell viability. Here, in xenografts generated with SH-SY5Y neuroblastoma cells stably expressing TRPM2 isoforms, growth of tumors expressing TRPM2-S was significantly reduced compared with tumors expressing TRPM2-L. Expression of hypoxia-inducible factor (HIF)-1/2α was significantly reduced in TRPM2-S-expressing tumor cells as was expression of target proteins regulated by HIF-1/2α including those involved in glycolysis (lactate dehydrogenaseAand enolase 2), oxidant stress (FOXO3a), angiogenesis (VEGF), mitophagy and mitochondrial function (BNIP3 and NDUFA4L2), and mitochondrial electron transport chain activity (cytochrome oxidase 4.1/4.2 in complex IV). The reduction in HIF-1/2α was mediated through both significantly reduced HIF-1/2α mRNA levels and increased levels of von Hippel-Lindau E3 ligase in TRPM2-S-expressing cells. Inhibition of TRPM2-L by pretreatment with clotrimazole or expression of TRPM2-S significantly increased sensitivity of cells to doxorubicin. Reduced survival of TRPM2-S-expressing cells after doxorubicin treatment was rescued by gain of HIF-1 or -2α function. These data suggest that TRPM2 activity is important for tumor growth and for cell viability and survival following doxorubicin treatment and that interference with TRPM2-L function may be a novel approach to reduce tumor growth through modulation of HIF-1/2α, mitochondrial function, and mitophagy.",
author = "Chen, {Shu Jen} and Hoffman, {Nicholas E.} and Santhanam Shanmughapriy and Lei Bao and Kerry Keefer and Kathleen Conrad and Salim Merali and Yoshinori Takahashi and Thomas Abraham and Iwona Hirschler-Laszkiewicz and Wang, {Ju Fang} and Zhang, {Xue Qian} and Jianliang Song and Carlos Barrero and Yuguang Shi and Kawasawa, {Yuka Imamura} and Michael Bayerl and Tianyu Sun and Mustafa Barbour and Wang, {Hong Gang} and Muniswamy Madesh and Cheung, {Joseph Y.} and Miller, {Barbara A.}",
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language = "English (US)",
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Chen, SJ, Hoffman, NE, Shanmughapriy, S, Bao, L, Keefer, K, Conrad, K, Merali, S, Takahashi, Y, Abraham, T, Hirschler-Laszkiewicz, I, Wang, JF, Zhang, XQ, Song, J, Barrero, C, Shi, Y, Kawasawa, YI, Bayerl, M, Sun, T, Barbour, M, Wang, HG, Madesh, M, Cheung, JY & Miller, BA 2014, 'A splice variant of the human ion channel TRPM2 modulates neuroblastoma tumor growth through hypoxia-inducible factor (HIF)-1/2α', Journal of Biological Chemistry, vol. 289, no. 52, pp. 36284-36302. https://doi.org/10.1074/jbc.M114.620922

A splice variant of the human ion channel TRPM2 modulates neuroblastoma tumor growth through hypoxia-inducible factor (HIF)-1/2α. / Chen, Shu Jen; Hoffman, Nicholas E.; Shanmughapriy, Santhanam; Bao, Lei; Keefer, Kerry; Conrad, Kathleen; Merali, Salim; Takahashi, Yoshinori; Abraham, Thomas; Hirschler-Laszkiewicz, Iwona; Wang, Ju Fang; Zhang, Xue Qian; Song, Jianliang; Barrero, Carlos; Shi, Yuguang; Kawasawa, Yuka Imamura; Bayerl, Michael; Sun, Tianyu; Barbour, Mustafa; Wang, Hong Gang; Madesh, Muniswamy; Cheung, Joseph Y.; Miller, Barbara A.

In: Journal of Biological Chemistry, Vol. 289, No. 52, 01.01.2014, p. 36284-36302.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A splice variant of the human ion channel TRPM2 modulates neuroblastoma tumor growth through hypoxia-inducible factor (HIF)-1/2α

AU - Chen, Shu Jen

AU - Hoffman, Nicholas E.

AU - Shanmughapriy, Santhanam

AU - Bao, Lei

AU - Keefer, Kerry

AU - Conrad, Kathleen

AU - Merali, Salim

AU - Takahashi, Yoshinori

AU - Abraham, Thomas

AU - Hirschler-Laszkiewicz, Iwona

AU - Wang, Ju Fang

AU - Zhang, Xue Qian

AU - Song, Jianliang

AU - Barrero, Carlos

AU - Shi, Yuguang

AU - Kawasawa, Yuka Imamura

AU - Bayerl, Michael

AU - Sun, Tianyu

AU - Barbour, Mustafa

AU - Wang, Hong Gang

AU - Madesh, Muniswamy

AU - Cheung, Joseph Y.

AU - Miller, Barbara A.

PY - 2014/1/1

Y1 - 2014/1/1

N2 - The calcium-permeable ion channel TRPM2 is highly expressed in a number of cancers. In neuroblastoma, full-length TRPM2 (TRPM2-L) protected cells from moderate oxidative stress through increased levels of forkhead box transcription factor 3a (FOXO3a) and superoxide dismutase 2. Cells expressing the dominant negative short isoform (TRPM2-S) had reduced FOXO3a and superoxide dismutase 2 levels, reduced calcium influx in response to oxidative stress, and enhanced reactive oxygen species, leading to decreased cell viability. Here, in xenografts generated with SH-SY5Y neuroblastoma cells stably expressing TRPM2 isoforms, growth of tumors expressing TRPM2-S was significantly reduced compared with tumors expressing TRPM2-L. Expression of hypoxia-inducible factor (HIF)-1/2α was significantly reduced in TRPM2-S-expressing tumor cells as was expression of target proteins regulated by HIF-1/2α including those involved in glycolysis (lactate dehydrogenaseAand enolase 2), oxidant stress (FOXO3a), angiogenesis (VEGF), mitophagy and mitochondrial function (BNIP3 and NDUFA4L2), and mitochondrial electron transport chain activity (cytochrome oxidase 4.1/4.2 in complex IV). The reduction in HIF-1/2α was mediated through both significantly reduced HIF-1/2α mRNA levels and increased levels of von Hippel-Lindau E3 ligase in TRPM2-S-expressing cells. Inhibition of TRPM2-L by pretreatment with clotrimazole or expression of TRPM2-S significantly increased sensitivity of cells to doxorubicin. Reduced survival of TRPM2-S-expressing cells after doxorubicin treatment was rescued by gain of HIF-1 or -2α function. These data suggest that TRPM2 activity is important for tumor growth and for cell viability and survival following doxorubicin treatment and that interference with TRPM2-L function may be a novel approach to reduce tumor growth through modulation of HIF-1/2α, mitochondrial function, and mitophagy.

AB - The calcium-permeable ion channel TRPM2 is highly expressed in a number of cancers. In neuroblastoma, full-length TRPM2 (TRPM2-L) protected cells from moderate oxidative stress through increased levels of forkhead box transcription factor 3a (FOXO3a) and superoxide dismutase 2. Cells expressing the dominant negative short isoform (TRPM2-S) had reduced FOXO3a and superoxide dismutase 2 levels, reduced calcium influx in response to oxidative stress, and enhanced reactive oxygen species, leading to decreased cell viability. Here, in xenografts generated with SH-SY5Y neuroblastoma cells stably expressing TRPM2 isoforms, growth of tumors expressing TRPM2-S was significantly reduced compared with tumors expressing TRPM2-L. Expression of hypoxia-inducible factor (HIF)-1/2α was significantly reduced in TRPM2-S-expressing tumor cells as was expression of target proteins regulated by HIF-1/2α including those involved in glycolysis (lactate dehydrogenaseAand enolase 2), oxidant stress (FOXO3a), angiogenesis (VEGF), mitophagy and mitochondrial function (BNIP3 and NDUFA4L2), and mitochondrial electron transport chain activity (cytochrome oxidase 4.1/4.2 in complex IV). The reduction in HIF-1/2α was mediated through both significantly reduced HIF-1/2α mRNA levels and increased levels of von Hippel-Lindau E3 ligase in TRPM2-S-expressing cells. Inhibition of TRPM2-L by pretreatment with clotrimazole or expression of TRPM2-S significantly increased sensitivity of cells to doxorubicin. Reduced survival of TRPM2-S-expressing cells after doxorubicin treatment was rescued by gain of HIF-1 or -2α function. These data suggest that TRPM2 activity is important for tumor growth and for cell viability and survival following doxorubicin treatment and that interference with TRPM2-L function may be a novel approach to reduce tumor growth through modulation of HIF-1/2α, mitochondrial function, and mitophagy.

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