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, Michael Bayerl, Tianyu Sun, Mustafa Barbour, Hong-Gang WangMuniswamy Madesh, Joseph Y. Cheung, Barbara Miller

Research output: Contribution to journalArticle

42 Citations (Scopus)

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

Fingerprint

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 ; Imamura, Yuka ; Bayerl, Michael ; Sun, Tianyu ; Barbour, Mustafa ; Wang, Hong-Gang ; Madesh, Muniswamy ; Cheung, Joseph Y. ; Miller, Barbara. / 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.
@article{fae4d9a7f6e24bbfb45a5162e980cdf2,
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 = "Shu-jen Chen 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 Yuka Imamura and Michael Bayerl and Tianyu Sun and Mustafa Barbour and Hong-Gang Wang and Muniswamy Madesh and Cheung, {Joseph Y.} and Barbara Miller",
year = "2014",
month = "1",
day = "1",
doi = "10.1074/jbc.M114.620922",
language = "English (US)",
volume = "289",
pages = "36284--36302",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "52",

}

Chen, S, 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, Imamura, Y, Bayerl, M, Sun, T, Barbour, M, Wang, H-G, Madesh, M, Cheung, JY & Miller, B 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; Imamura, Yuka; Bayerl, Michael; Sun, Tianyu; Barbour, Mustafa; Wang, Hong-Gang; Madesh, Muniswamy; Cheung, Joseph Y.; Miller, Barbara.

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 - Imamura, Yuka

AU - Bayerl, Michael

AU - Sun, Tianyu

AU - Barbour, Mustafa

AU - Wang, Hong-Gang

AU - Madesh, Muniswamy

AU - Cheung, Joseph Y.

AU - Miller, Barbara

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.

UR - http://www.scopus.com/inward/record.url?scp=84937242801&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84937242801&partnerID=8YFLogxK

U2 - 10.1074/jbc.M114.620922

DO - 10.1074/jbc.M114.620922

M3 - Article

C2 - 25391657

AN - SCOPUS:84937242801

VL - 289

SP - 36284

EP - 36302

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 52

ER -