Reexpression of LGI1 in glioma cells results in dysregulation of genes implicated in the canonical axon guidance pathway

Padmaja Kunapuli, Ken Lo, Lesleyann Hawthorn, John Kenneth Cowell

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The LGI1 gene suppresses invasion in glioma cells and predisposes to epilepsy. In a gene expression array comparison between parental cells and T98G cell clones forced to express LGI1, we demonstrate that the canonical axon guidance pathway is the most significantly affected. In particular, aspects of axon guidance that involve reorganization of the actin cytoskeleton, which is also involved in cell movement and invasion, were affected. Analysis of actin fiber organization using fluorescence microscopy demonstrated that different T98G cell clones expressing the exogenous LGI1 gene show high levels of stress fibers compared with controls. Since stress fiber formation is associated with loss of cell mobility, we used scratch wound assays to demonstrate that LGI1-expressing clones show a significant reduction in cell mobility. LGI1 reexpression also resulted in loss of the PDGFRA and EGFR proteins, suggesting a rapid turnover of these receptors despite increased mRNA levels for PDGFRA. LGI1 suppression of invasion is associated with loss of ERK/MAPK1 activation. LGI1 is a secreted protein, and when the culture supernatant from cells expressing FLAG- and GFP-tagged proteins were applied to parental T98G cells, ERK/MAPK1 phosphorylation and cell mobility was suppressed, demonstrating that the LGI1 protein acts as a suppressive agent for cell movement in this assay. These observations support a previous suggestion that LGI1 can reduce cellular invasion in in vitro assays and, as a secreted agent, may be developed as a means of treating metastatic cancer. In addition, this observation provides a mechanistic link for LGI1's common role in metastasis and epilepsy development.

Original languageEnglish (US)
Pages (from-to)93-100
Number of pages8
JournalGenomics
Volume95
Issue number2
DOIs
StatePublished - Feb 1 2010

Fingerprint

Glioma
Genes
Stress Fibers
Clone Cells
Cell Movement
Epilepsy
Proteins
Actin Cytoskeleton
Fluorescence Microscopy
Axon Guidance
Actins
Cell Culture Techniques
Phosphorylation
Neoplasm Metastasis
Gene Expression
Messenger RNA
Wounds and Injuries
Neoplasms

Keywords

  • Actin cytoskeleton
  • Axon guidance
  • Expression microarray
  • Invasion
  • Lgi1

ASJC Scopus subject areas

  • Genetics

Cite this

Reexpression of LGI1 in glioma cells results in dysregulation of genes implicated in the canonical axon guidance pathway. / Kunapuli, Padmaja; Lo, Ken; Hawthorn, Lesleyann; Cowell, John Kenneth.

In: Genomics, Vol. 95, No. 2, 01.02.2010, p. 93-100.

Research output: Contribution to journalArticle

@article{43a0a309fe0c48c98f18a417daf337ca,
title = "Reexpression of LGI1 in glioma cells results in dysregulation of genes implicated in the canonical axon guidance pathway",
abstract = "The LGI1 gene suppresses invasion in glioma cells and predisposes to epilepsy. In a gene expression array comparison between parental cells and T98G cell clones forced to express LGI1, we demonstrate that the canonical axon guidance pathway is the most significantly affected. In particular, aspects of axon guidance that involve reorganization of the actin cytoskeleton, which is also involved in cell movement and invasion, were affected. Analysis of actin fiber organization using fluorescence microscopy demonstrated that different T98G cell clones expressing the exogenous LGI1 gene show high levels of stress fibers compared with controls. Since stress fiber formation is associated with loss of cell mobility, we used scratch wound assays to demonstrate that LGI1-expressing clones show a significant reduction in cell mobility. LGI1 reexpression also resulted in loss of the PDGFRA and EGFR proteins, suggesting a rapid turnover of these receptors despite increased mRNA levels for PDGFRA. LGI1 suppression of invasion is associated with loss of ERK/MAPK1 activation. LGI1 is a secreted protein, and when the culture supernatant from cells expressing FLAG- and GFP-tagged proteins were applied to parental T98G cells, ERK/MAPK1 phosphorylation and cell mobility was suppressed, demonstrating that the LGI1 protein acts as a suppressive agent for cell movement in this assay. These observations support a previous suggestion that LGI1 can reduce cellular invasion in in vitro assays and, as a secreted agent, may be developed as a means of treating metastatic cancer. In addition, this observation provides a mechanistic link for LGI1's common role in metastasis and epilepsy development.",
keywords = "Actin cytoskeleton, Axon guidance, Expression microarray, Invasion, Lgi1",
author = "Padmaja Kunapuli and Ken Lo and Lesleyann Hawthorn and Cowell, {John Kenneth}",
year = "2010",
month = "2",
day = "1",
doi = "10.1016/j.ygeno.2009.10.001",
language = "English (US)",
volume = "95",
pages = "93--100",
journal = "Genomics",
issn = "0888-7543",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Reexpression of LGI1 in glioma cells results in dysregulation of genes implicated in the canonical axon guidance pathway

AU - Kunapuli, Padmaja

AU - Lo, Ken

AU - Hawthorn, Lesleyann

AU - Cowell, John Kenneth

PY - 2010/2/1

Y1 - 2010/2/1

N2 - The LGI1 gene suppresses invasion in glioma cells and predisposes to epilepsy. In a gene expression array comparison between parental cells and T98G cell clones forced to express LGI1, we demonstrate that the canonical axon guidance pathway is the most significantly affected. In particular, aspects of axon guidance that involve reorganization of the actin cytoskeleton, which is also involved in cell movement and invasion, were affected. Analysis of actin fiber organization using fluorescence microscopy demonstrated that different T98G cell clones expressing the exogenous LGI1 gene show high levels of stress fibers compared with controls. Since stress fiber formation is associated with loss of cell mobility, we used scratch wound assays to demonstrate that LGI1-expressing clones show a significant reduction in cell mobility. LGI1 reexpression also resulted in loss of the PDGFRA and EGFR proteins, suggesting a rapid turnover of these receptors despite increased mRNA levels for PDGFRA. LGI1 suppression of invasion is associated with loss of ERK/MAPK1 activation. LGI1 is a secreted protein, and when the culture supernatant from cells expressing FLAG- and GFP-tagged proteins were applied to parental T98G cells, ERK/MAPK1 phosphorylation and cell mobility was suppressed, demonstrating that the LGI1 protein acts as a suppressive agent for cell movement in this assay. These observations support a previous suggestion that LGI1 can reduce cellular invasion in in vitro assays and, as a secreted agent, may be developed as a means of treating metastatic cancer. In addition, this observation provides a mechanistic link for LGI1's common role in metastasis and epilepsy development.

AB - The LGI1 gene suppresses invasion in glioma cells and predisposes to epilepsy. In a gene expression array comparison between parental cells and T98G cell clones forced to express LGI1, we demonstrate that the canonical axon guidance pathway is the most significantly affected. In particular, aspects of axon guidance that involve reorganization of the actin cytoskeleton, which is also involved in cell movement and invasion, were affected. Analysis of actin fiber organization using fluorescence microscopy demonstrated that different T98G cell clones expressing the exogenous LGI1 gene show high levels of stress fibers compared with controls. Since stress fiber formation is associated with loss of cell mobility, we used scratch wound assays to demonstrate that LGI1-expressing clones show a significant reduction in cell mobility. LGI1 reexpression also resulted in loss of the PDGFRA and EGFR proteins, suggesting a rapid turnover of these receptors despite increased mRNA levels for PDGFRA. LGI1 suppression of invasion is associated with loss of ERK/MAPK1 activation. LGI1 is a secreted protein, and when the culture supernatant from cells expressing FLAG- and GFP-tagged proteins were applied to parental T98G cells, ERK/MAPK1 phosphorylation and cell mobility was suppressed, demonstrating that the LGI1 protein acts as a suppressive agent for cell movement in this assay. These observations support a previous suggestion that LGI1 can reduce cellular invasion in in vitro assays and, as a secreted agent, may be developed as a means of treating metastatic cancer. In addition, this observation provides a mechanistic link for LGI1's common role in metastasis and epilepsy development.

KW - Actin cytoskeleton

KW - Axon guidance

KW - Expression microarray

KW - Invasion

KW - Lgi1

UR - http://www.scopus.com/inward/record.url?scp=75749151214&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=75749151214&partnerID=8YFLogxK

U2 - 10.1016/j.ygeno.2009.10.001

DO - 10.1016/j.ygeno.2009.10.001

M3 - Article

VL - 95

SP - 93

EP - 100

JO - Genomics

JF - Genomics

SN - 0888-7543

IS - 2

ER -