Silencing of the microtubule-associated proteins doublecortin-like and doublecortin-like kinase-long induces apoptosis in neuroblastoma cells

Carla S Verissimo; Jan J Molenaar; John Meerman; Jordi Carreras Puigvert; Fieke Lamers; Jan Koster; Erik H J Danen; Bob van de Water; Rogier Versteeg; Carlos P Fitzsimons; Erno Vreugdenhil

doi:10.1677/ERC-09-0301

Silencing of the microtubule-associated proteins doublecortin-like and doublecortin-like kinase-long induces apoptosis in neuroblastoma cells

Carla S Verissimo
, Jan J Molenaar
, John Meerman
, Jordi Carreras Puigvert
, Fieke Lamers
, Jan Koster
, Erik H J Danen
, Bob van de Water
, Rogier Versteeg
, Carlos P Fitzsimons
, Erno Vreugdenhil

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Doublecortin-like kinase-long (DCLK-long) and doublecortin-like (DCL) are two splice variants of DCLK gene. DCL and DCLK-long are microtubule-associated proteins with specific expression in proliferative neural progenitor cells. We have tested the hypothesis that knockdown of DCL/DCLK-long by RNA interference technology will induce cell death in neuroblastoma (NB) cells. First, we analyzed the expression of DCL and DCLK-long in several human neuroblastic tumors, other tumors, and normal tissues, revealing high expression of both DCL and DCLK-long in NB and glioma. Secondly, gene expression profiling revealed numerous differentially expressed genes indicating apoptosis induction after DCL/DCLK-long knockdown in NB cells. Finally, apoptosis was confirmed by time-lapse imaging of phosphatidylserine translocation, caspase-3 activation, live/dead double staining assays, and fluorescence-activated cell sorting. Together, our results suggest that silencing DCL/DCLK-long induces apoptosis in NB cells.

Original language	English
Pages (from-to)	399-414
Number of pages	16
Journal	Endocrine-Related Cancer
Volume	17
Issue number	2
DOIs	https://doi.org/10.1677/ERC-09-0301
Publication status	Published - Jun 2010
Externally published	Yes

Keywords

Animals
Apoptosis/genetics
Blotting, Western
Caspase 3/genetics
Cell Count
Cell Line, Tumor
Cells, Cultured
Flow Cytometry
Gene Expression Profiling
Humans
Image Processing, Computer-Assisted
Intracellular Signaling Peptides and Proteins/genetics
Mice
Neurons/metabolism
Protein Isoforms/genetics
Protein-Serine-Threonine Kinases/genetics
RNA Interference
RNA, Messenger/genetics
RNA, Small Interfering
Reverse Transcriptase Polymerase Chain Reaction
Stem Cells/cytology

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10.1677/ERC-09-0301

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Verissimo, C. S., Molenaar, J. J., Meerman, J., Puigvert, J. C., Lamers, F., Koster, J., Danen, E. H. J., van de Water, B., Versteeg, R., Fitzsimons, C. P., & Vreugdenhil, E. (2010). Silencing of the microtubule-associated proteins doublecortin-like and doublecortin-like kinase-long induces apoptosis in neuroblastoma cells. Endocrine-Related Cancer, 17(2), 399-414. https://doi.org/10.1677/ERC-09-0301

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title = "Silencing of the microtubule-associated proteins doublecortin-like and doublecortin-like kinase-long induces apoptosis in neuroblastoma cells",

abstract = "Doublecortin-like kinase-long (DCLK-long) and doublecortin-like (DCL) are two splice variants of DCLK gene. DCL and DCLK-long are microtubule-associated proteins with specific expression in proliferative neural progenitor cells. We have tested the hypothesis that knockdown of DCL/DCLK-long by RNA interference technology will induce cell death in neuroblastoma (NB) cells. First, we analyzed the expression of DCL and DCLK-long in several human neuroblastic tumors, other tumors, and normal tissues, revealing high expression of both DCL and DCLK-long in NB and glioma. Secondly, gene expression profiling revealed numerous differentially expressed genes indicating apoptosis induction after DCL/DCLK-long knockdown in NB cells. Finally, apoptosis was confirmed by time-lapse imaging of phosphatidylserine translocation, caspase-3 activation, live/dead double staining assays, and fluorescence-activated cell sorting. Together, our results suggest that silencing DCL/DCLK-long induces apoptosis in NB cells.",

keywords = "Animals, Apoptosis/genetics, Blotting, Western, Caspase 3/genetics, Cell Count, Cell Line, Tumor, Cells, Cultured, Flow Cytometry, Gene Expression Profiling, Humans, Image Processing, Computer-Assisted, Intracellular Signaling Peptides and Proteins/genetics, Mice, Neurons/metabolism, Protein Isoforms/genetics, Protein-Serine-Threonine Kinases/genetics, RNA Interference, RNA, Messenger/genetics, RNA, Small Interfering, Reverse Transcriptase Polymerase Chain Reaction, Stem Cells/cytology",

author = "Verissimo, \{Carla S\} and Molenaar, \{Jan J\} and John Meerman and Puigvert, \{Jordi Carreras\} and Fieke Lamers and Jan Koster and Danen, \{Erik H J\} and \{van de Water\}, Bob and Rogier Versteeg and Fitzsimons, \{Carlos P\} and Erno Vreugdenhil",

year = "2010",

month = jun,

doi = "10.1677/ERC-09-0301",

language = "English",

volume = "17",

pages = "399--414",

journal = "Endocrine-Related Cancer",

issn = "1351-0088",

publisher = "BioScientifica Ltd.",

number = "2",

}

Verissimo, CS, Molenaar, JJ, Meerman, J, Puigvert, JC, Lamers, F, Koster, J, Danen, EHJ, van de Water, B, Versteeg, R, Fitzsimons, CP & Vreugdenhil, E 2010, 'Silencing of the microtubule-associated proteins doublecortin-like and doublecortin-like kinase-long induces apoptosis in neuroblastoma cells', Endocrine-Related Cancer, vol. 17, no. 2, pp. 399-414. https://doi.org/10.1677/ERC-09-0301

TY - JOUR

T1 - Silencing of the microtubule-associated proteins doublecortin-like and doublecortin-like kinase-long induces apoptosis in neuroblastoma cells

AU - Verissimo, Carla S

AU - Molenaar, Jan J

AU - Meerman, John

AU - Puigvert, Jordi Carreras

AU - Lamers, Fieke

AU - Koster, Jan

AU - Danen, Erik H J

AU - van de Water, Bob

AU - Versteeg, Rogier

AU - Fitzsimons, Carlos P

AU - Vreugdenhil, Erno

PY - 2010/6

Y1 - 2010/6

N2 - Doublecortin-like kinase-long (DCLK-long) and doublecortin-like (DCL) are two splice variants of DCLK gene. DCL and DCLK-long are microtubule-associated proteins with specific expression in proliferative neural progenitor cells. We have tested the hypothesis that knockdown of DCL/DCLK-long by RNA interference technology will induce cell death in neuroblastoma (NB) cells. First, we analyzed the expression of DCL and DCLK-long in several human neuroblastic tumors, other tumors, and normal tissues, revealing high expression of both DCL and DCLK-long in NB and glioma. Secondly, gene expression profiling revealed numerous differentially expressed genes indicating apoptosis induction after DCL/DCLK-long knockdown in NB cells. Finally, apoptosis was confirmed by time-lapse imaging of phosphatidylserine translocation, caspase-3 activation, live/dead double staining assays, and fluorescence-activated cell sorting. Together, our results suggest that silencing DCL/DCLK-long induces apoptosis in NB cells.

AB - Doublecortin-like kinase-long (DCLK-long) and doublecortin-like (DCL) are two splice variants of DCLK gene. DCL and DCLK-long are microtubule-associated proteins with specific expression in proliferative neural progenitor cells. We have tested the hypothesis that knockdown of DCL/DCLK-long by RNA interference technology will induce cell death in neuroblastoma (NB) cells. First, we analyzed the expression of DCL and DCLK-long in several human neuroblastic tumors, other tumors, and normal tissues, revealing high expression of both DCL and DCLK-long in NB and glioma. Secondly, gene expression profiling revealed numerous differentially expressed genes indicating apoptosis induction after DCL/DCLK-long knockdown in NB cells. Finally, apoptosis was confirmed by time-lapse imaging of phosphatidylserine translocation, caspase-3 activation, live/dead double staining assays, and fluorescence-activated cell sorting. Together, our results suggest that silencing DCL/DCLK-long induces apoptosis in NB cells.

KW - Animals

KW - Apoptosis/genetics

KW - Blotting, Western

KW - Caspase 3/genetics

KW - Cell Count

KW - Cell Line, Tumor

KW - Cells, Cultured

KW - Flow Cytometry

KW - Gene Expression Profiling

KW - Humans

KW - Image Processing, Computer-Assisted

KW - Intracellular Signaling Peptides and Proteins/genetics

KW - Mice

KW - Neurons/metabolism

KW - Protein Isoforms/genetics

KW - Protein-Serine-Threonine Kinases/genetics

KW - RNA Interference

KW - RNA, Messenger/genetics

KW - RNA, Small Interfering

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Stem Cells/cytology

U2 - 10.1677/ERC-09-0301

DO - 10.1677/ERC-09-0301

M3 - Article

C2 - 20228126

SN - 1351-0088

VL - 17

SP - 399

EP - 414

JO - Endocrine-Related Cancer

JF - Endocrine-Related Cancer

IS - 2

ER -

Silencing of the microtubule-associated proteins doublecortin-like and doublecortin-like kinase-long induces apoptosis in neuroblastoma cells

Abstract

Keywords

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