Altered calcium regulation by thermosensitive transient receptor potential channels in etoposide-resistant WERI-Rb1 retinoblastoma cells

Stefan Mergler; Yating Cheng; Sergej Skosyrski; Fabian Garreis; Piotr Pietrzak; Norbert Kociok; Abhilash Dwarakanath; Peter S. Reinach; Vinodh Kakkassery

doi:10.1016/j.exer.2011.12.002

Altered calcium regulation by thermosensitive transient receptor potential channels in etoposide-resistant WERI-Rb1 retinoblastoma cells

Stefan Mergler^*, Yating Cheng, Sergej Skosyrski, Fabian Garreis, Piotr Pietrzak, Norbert Kociok, Abhilash Dwarakanath, Peter S. Reinach, Vinodh Kakkassery

^*Corresponding author for this work

Department of Ophtalmology

33 Citations (Scopus)

Abstract

Differences in transient receptor potential (TRP) and cannabinoid receptor type 1 (CB1) expression levels can serve as prognostic factors for retinoblastoma (RB) tumor progression. We hypothesized in RB tissue that such differences are also indicators of whether or not they are sensitive to etoposide. Accordingly, we compared in malignant etoposide-sensitive and etoposide-resistant WERI-Rb1 cells TRPV1, TRPM8 and TRPA1 subtype and CB1 gene expression pattern levels and accompanying functional activity using quantitative real-time RT-PCR, immunohistochemistry, immunofluorescence microscopy, calcium imaging as well as patch-clamp technology. Gene expression patterns were evaluated in enucleated human RB tissues (n=4). Both etoposide-resistant and etoposide-sensitive WERI-Rb1 cells expressed all of the aforementioned channels based on responses to known activators and thermal challenges. However, TRPA1 was absent in the etoposide-resistant counterpart. Even though both types of RB cells express TRPV1 as well as TRPM8 and CB1, the capsaicin (50μM) (CAP)-induced Ca ²⁺ rise caused by TRPV1 activation was prompt and transient only in etoposide-resistant RB cells (n=8). In this cell type, the inability of CB1 activation (10μM WIN) to suppress Ca ²⁺ responses to CAP (50μM; n=4) may be attributable to the absence of TRPA1 gene expression. Therefore, using genetic approaches to upregulate TRPA1 expression could provide a means to induce etoposide sensitivity and suppress RB cell tumorigenesis.

Original language	English
Journal	Experimental Eye Research
Volume	94
Issue number	1
Pages (from-to)	157-173
Number of pages	17
ISSN	0014-4835
DOIs	https://doi.org/10.1016/j.exer.2011.12.002
Publication status	Published - 01.2012

Funding

Contract grant sponsor: Berliner Sonnenfeld-Stiftung , Contract grant number: 89745052 . Contract grant sponsor: Jackstädt-Stiftung Wuppertal , Contract grant number: S 134-10.063 . The authors thank Gabriele Fels and Sylvia Metzner for technical assistance as well as Dr. Irina Semkova for helpful discussions. In addition, the authors also thank Dr. Gregor Willerding-Beaucamp and Dr. Aline Riechardt for supporting the evaluation of immunofluorescence microscopy of normal retina and RB tissue. Dr. Harald Stephan and colleagues kindly provided the RB cell lines. Dr. Vinodh Kakkassery was supported by Jackstädt Stiftung (Wuppertal, Germany). The planar patch-clamp equipment was supported in part by Sonnenfeld-Stiftung (Berlin, Germany). Finally, we also appreciate the technical assistance provided by the fellow student BSc Nefeli Slavi (Int. Graduate Program in Medical Neurosciences) during her lab practice.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being
SDG 9 Industry, Innovation, and Infrastructure

Research Areas and Centers

Research Area: Luebeck Integrated Oncology Network (LION)

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@article{e0d00464b325404abbaf76cf8927df8a,

title = "Altered calcium regulation by thermosensitive transient receptor potential channels in etoposide-resistant WERI-Rb1 retinoblastoma cells",

abstract = "Differences in transient receptor potential (TRP) and cannabinoid receptor type 1 (CB1) expression levels can serve as prognostic factors for retinoblastoma (RB) tumor progression. We hypothesized in RB tissue that such differences are also indicators of whether or not they are sensitive to etoposide. Accordingly, we compared in malignant etoposide-sensitive and etoposide-resistant WERI-Rb1 cells TRPV1, TRPM8 and TRPA1 subtype and CB1 gene expression pattern levels and accompanying functional activity using quantitative real-time RT-PCR, immunohistochemistry, immunofluorescence microscopy, calcium imaging as well as patch-clamp technology. Gene expression patterns were evaluated in enucleated human RB tissues (n=4). Both etoposide-resistant and etoposide-sensitive WERI-Rb1 cells expressed all of the aforementioned channels based on responses to known activators and thermal challenges. However, TRPA1 was absent in the etoposide-resistant counterpart. Even though both types of RB cells express TRPV1 as well as TRPM8 and CB1, the capsaicin (50μM) (CAP)-induced Ca 2+ rise caused by TRPV1 activation was prompt and transient only in etoposide-resistant RB cells (n=8). In this cell type, the inability of CB1 activation (10μM WIN) to suppress Ca 2+ responses to CAP (50μM; n=4) may be attributable to the absence of TRPA1 gene expression. Therefore, using genetic approaches to upregulate TRPA1 expression could provide a means to induce etoposide sensitivity and suppress RB cell tumorigenesis.",

author = "Stefan Mergler and Yating Cheng and Sergej Skosyrski and Fabian Garreis and Piotr Pietrzak and Norbert Kociok and Abhilash Dwarakanath and Reinach, \{Peter S.\} and Vinodh Kakkassery",

note = "Funding Information: Contract grant sponsor: Berliner Sonnenfeld-Stiftung , Contract grant number: 89745052 . Contract grant sponsor: Jackst{\"a}dt-Stiftung Wuppertal , Contract grant number: S 134-10.063 . Funding Information: The authors thank Gabriele Fels and Sylvia Metzner for technical assistance as well as Dr. Irina Semkova for helpful discussions. In addition, the authors also thank Dr. Gregor Willerding-Beaucamp and Dr. Aline Riechardt for supporting the evaluation of immunofluorescence microscopy of normal retina and RB tissue. Dr. Harald Stephan and colleagues kindly provided the RB cell lines. Dr. Vinodh Kakkassery was supported by Jackst{\"a}dt Stiftung (Wuppertal, Germany). The planar patch-clamp equipment was supported in part by Sonnenfeld-Stiftung (Berlin, Germany). Finally, we also appreciate the technical assistance provided by the fellow student BSc Nefeli Slavi (Int. Graduate Program in Medical Neurosciences) during her lab practice. Copyright: Copyright 2012 Elsevier B.V., All rights reserved.",

year = "2012",

month = jan,

doi = "10.1016/j.exer.2011.12.002",

language = "English",

volume = "94",

pages = "157--173",

journal = "Experimental Eye Research",

issn = "0014-4835",

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T1 - Altered calcium regulation by thermosensitive transient receptor potential channels in etoposide-resistant WERI-Rb1 retinoblastoma cells

AU - Mergler, Stefan

AU - Cheng, Yating

AU - Skosyrski, Sergej

AU - Garreis, Fabian

AU - Pietrzak, Piotr

AU - Kociok, Norbert

AU - Dwarakanath, Abhilash

AU - Reinach, Peter S.

AU - Kakkassery, Vinodh

N1 - Funding Information: Contract grant sponsor: Berliner Sonnenfeld-Stiftung , Contract grant number: 89745052 . Contract grant sponsor: Jackstädt-Stiftung Wuppertal , Contract grant number: S 134-10.063 . Funding Information: The authors thank Gabriele Fels and Sylvia Metzner for technical assistance as well as Dr. Irina Semkova for helpful discussions. In addition, the authors also thank Dr. Gregor Willerding-Beaucamp and Dr. Aline Riechardt for supporting the evaluation of immunofluorescence microscopy of normal retina and RB tissue. Dr. Harald Stephan and colleagues kindly provided the RB cell lines. Dr. Vinodh Kakkassery was supported by Jackstädt Stiftung (Wuppertal, Germany). The planar patch-clamp equipment was supported in part by Sonnenfeld-Stiftung (Berlin, Germany). Finally, we also appreciate the technical assistance provided by the fellow student BSc Nefeli Slavi (Int. Graduate Program in Medical Neurosciences) during her lab practice. Copyright: Copyright 2012 Elsevier B.V., All rights reserved.

PY - 2012/1

Y1 - 2012/1

N2 - Differences in transient receptor potential (TRP) and cannabinoid receptor type 1 (CB1) expression levels can serve as prognostic factors for retinoblastoma (RB) tumor progression. We hypothesized in RB tissue that such differences are also indicators of whether or not they are sensitive to etoposide. Accordingly, we compared in malignant etoposide-sensitive and etoposide-resistant WERI-Rb1 cells TRPV1, TRPM8 and TRPA1 subtype and CB1 gene expression pattern levels and accompanying functional activity using quantitative real-time RT-PCR, immunohistochemistry, immunofluorescence microscopy, calcium imaging as well as patch-clamp technology. Gene expression patterns were evaluated in enucleated human RB tissues (n=4). Both etoposide-resistant and etoposide-sensitive WERI-Rb1 cells expressed all of the aforementioned channels based on responses to known activators and thermal challenges. However, TRPA1 was absent in the etoposide-resistant counterpart. Even though both types of RB cells express TRPV1 as well as TRPM8 and CB1, the capsaicin (50μM) (CAP)-induced Ca 2+ rise caused by TRPV1 activation was prompt and transient only in etoposide-resistant RB cells (n=8). In this cell type, the inability of CB1 activation (10μM WIN) to suppress Ca 2+ responses to CAP (50μM; n=4) may be attributable to the absence of TRPA1 gene expression. Therefore, using genetic approaches to upregulate TRPA1 expression could provide a means to induce etoposide sensitivity and suppress RB cell tumorigenesis.

AB - Differences in transient receptor potential (TRP) and cannabinoid receptor type 1 (CB1) expression levels can serve as prognostic factors for retinoblastoma (RB) tumor progression. We hypothesized in RB tissue that such differences are also indicators of whether or not they are sensitive to etoposide. Accordingly, we compared in malignant etoposide-sensitive and etoposide-resistant WERI-Rb1 cells TRPV1, TRPM8 and TRPA1 subtype and CB1 gene expression pattern levels and accompanying functional activity using quantitative real-time RT-PCR, immunohistochemistry, immunofluorescence microscopy, calcium imaging as well as patch-clamp technology. Gene expression patterns were evaluated in enucleated human RB tissues (n=4). Both etoposide-resistant and etoposide-sensitive WERI-Rb1 cells expressed all of the aforementioned channels based on responses to known activators and thermal challenges. However, TRPA1 was absent in the etoposide-resistant counterpart. Even though both types of RB cells express TRPV1 as well as TRPM8 and CB1, the capsaicin (50μM) (CAP)-induced Ca 2+ rise caused by TRPV1 activation was prompt and transient only in etoposide-resistant RB cells (n=8). In this cell type, the inability of CB1 activation (10μM WIN) to suppress Ca 2+ responses to CAP (50μM; n=4) may be attributable to the absence of TRPA1 gene expression. Therefore, using genetic approaches to upregulate TRPA1 expression could provide a means to induce etoposide sensitivity and suppress RB cell tumorigenesis.

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U2 - 10.1016/j.exer.2011.12.002

DO - 10.1016/j.exer.2011.12.002

M3 - Journal articles

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AN - SCOPUS:84855854181

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JO - Experimental Eye Research

JF - Experimental Eye Research

IS - 1

ER -

Altered calcium regulation by thermosensitive transient receptor potential channels in etoposide-resistant WERI-Rb1 retinoblastoma cells

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