Epigenetic inactivation of laminin-5-encoding genes in lung cancers

Ubaradka G. Sathyanarayana, Shinichi Toyooka, Asha Padar, Takashi Takahashi, Elizabeth Brambilla, John D. Minna, Adi F. Gazdar

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Purpose: We investigated the loss of expression of three laminin-5 (LN5)-encoding genes in lung cancer cell lines and elucidated the mechanism of inactivation of the genes in lung cancer cell lines and tumors. Experimental Design: We examined the expression of LN5-encoding genes by reverse transcription-PCR in 49 lung cancer cell lines. To elucidate the mechanism of gene silencing, we treated expression-negative cell lines (two for each gene) with a demethylating agent and examined the restoration of expression by reverse transcription-PCR. We dissected out the methylation patterns of CpG sites unique to the promoter regions of LN5-encoding genes by bisulfite genomic sequencing of expression-negative cell lines. We designed methylation-specific primers and validated the methylation status of the promoter regions in lung cancer cell lines using methylation-specific PCR. We further studied the methylation patterns of primary non-small cell lung cancer [NSCLC (n = 36)], small cell lung cancer [SCLC (n = 26)], and carcinoids (n = 24) tumors. Results: We observed frequent losses of expression in NSCLC (20-60%) and SCLC (65-86%) cell lines. Expression of one or more genes was lost in 90% of SCLC cell lines and 65% of NSCLC cell lines. Treatment of expression-negative cell lines with demethylating agent restored expression in all of the cases. Methylation of LN5-encoding genes was present more frequently in SCLC cell lines (60-80%) than in NSCLC cell lines (15-60%), and at least one gene was methylated in 95% of SCLC and 60% of NSCLC cell lines. The concordances between loss of expression and methylation in 40 lung cancer cell lines for the three genes (90-95%) were statistically significant. Methylation was more frequent in SCLC tumors (58-77%) than in NSCLC tumors (22-42%) and carcinoids (13-33%), and at least one gene was methylated in 92% of SCLC tumors, 47% of NSCLC tumors, and 33% of carcinoids. Conclusions: Our results demonstrate frequent epigenetic inactivation of LN5-encoding genes in lung cancers, and these findings are of biological interest and are potentially of clinical importance.

Original languageEnglish
Pages (from-to)2665-2672
Number of pages8
JournalClinical Cancer Research
Volume9
Issue number7
Publication statusPublished - Jul 1 2003
Externally publishedYes

Fingerprint

Epigenomics
Lung Neoplasms
Cell Line
Methylation
Genes
Carcinoid Tumor
Gene Silencing
Genetic Promoter Regions
kalinin
Polymerase Chain Reaction
Reverse Transcription
Neoplasms
Small Cell Lung Carcinoma
Tumor Cell Line
Non-Small Cell Lung Carcinoma
Research Design

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Sathyanarayana, U. G., Toyooka, S., Padar, A., Takahashi, T., Brambilla, E., Minna, J. D., & Gazdar, A. F. (2003). Epigenetic inactivation of laminin-5-encoding genes in lung cancers. Clinical Cancer Research, 9(7), 2665-2672.

Epigenetic inactivation of laminin-5-encoding genes in lung cancers. / Sathyanarayana, Ubaradka G.; Toyooka, Shinichi; Padar, Asha; Takahashi, Takashi; Brambilla, Elizabeth; Minna, John D.; Gazdar, Adi F.

In: Clinical Cancer Research, Vol. 9, No. 7, 01.07.2003, p. 2665-2672.

Research output: Contribution to journalArticle

Sathyanarayana, UG, Toyooka, S, Padar, A, Takahashi, T, Brambilla, E, Minna, JD & Gazdar, AF 2003, 'Epigenetic inactivation of laminin-5-encoding genes in lung cancers', Clinical Cancer Research, vol. 9, no. 7, pp. 2665-2672.
Sathyanarayana UG, Toyooka S, Padar A, Takahashi T, Brambilla E, Minna JD et al. Epigenetic inactivation of laminin-5-encoding genes in lung cancers. Clinical Cancer Research. 2003 Jul 1;9(7):2665-2672.
Sathyanarayana, Ubaradka G. ; Toyooka, Shinichi ; Padar, Asha ; Takahashi, Takashi ; Brambilla, Elizabeth ; Minna, John D. ; Gazdar, Adi F. / Epigenetic inactivation of laminin-5-encoding genes in lung cancers. In: Clinical Cancer Research. 2003 ; Vol. 9, No. 7. pp. 2665-2672.
@article{473fec0c433a455db2792e47734e47bb,
title = "Epigenetic inactivation of laminin-5-encoding genes in lung cancers",
abstract = "Purpose: We investigated the loss of expression of three laminin-5 (LN5)-encoding genes in lung cancer cell lines and elucidated the mechanism of inactivation of the genes in lung cancer cell lines and tumors. Experimental Design: We examined the expression of LN5-encoding genes by reverse transcription-PCR in 49 lung cancer cell lines. To elucidate the mechanism of gene silencing, we treated expression-negative cell lines (two for each gene) with a demethylating agent and examined the restoration of expression by reverse transcription-PCR. We dissected out the methylation patterns of CpG sites unique to the promoter regions of LN5-encoding genes by bisulfite genomic sequencing of expression-negative cell lines. We designed methylation-specific primers and validated the methylation status of the promoter regions in lung cancer cell lines using methylation-specific PCR. We further studied the methylation patterns of primary non-small cell lung cancer [NSCLC (n = 36)], small cell lung cancer [SCLC (n = 26)], and carcinoids (n = 24) tumors. Results: We observed frequent losses of expression in NSCLC (20-60{\%}) and SCLC (65-86{\%}) cell lines. Expression of one or more genes was lost in 90{\%} of SCLC cell lines and 65{\%} of NSCLC cell lines. Treatment of expression-negative cell lines with demethylating agent restored expression in all of the cases. Methylation of LN5-encoding genes was present more frequently in SCLC cell lines (60-80{\%}) than in NSCLC cell lines (15-60{\%}), and at least one gene was methylated in 95{\%} of SCLC and 60{\%} of NSCLC cell lines. The concordances between loss of expression and methylation in 40 lung cancer cell lines for the three genes (90-95{\%}) were statistically significant. Methylation was more frequent in SCLC tumors (58-77{\%}) than in NSCLC tumors (22-42{\%}) and carcinoids (13-33{\%}), and at least one gene was methylated in 92{\%} of SCLC tumors, 47{\%} of NSCLC tumors, and 33{\%} of carcinoids. Conclusions: Our results demonstrate frequent epigenetic inactivation of LN5-encoding genes in lung cancers, and these findings are of biological interest and are potentially of clinical importance.",
author = "Sathyanarayana, {Ubaradka G.} and Shinichi Toyooka and Asha Padar and Takashi Takahashi and Elizabeth Brambilla and Minna, {John D.} and Gazdar, {Adi F.}",
year = "2003",
month = "7",
day = "1",
language = "English",
volume = "9",
pages = "2665--2672",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "7",

}

TY - JOUR

T1 - Epigenetic inactivation of laminin-5-encoding genes in lung cancers

AU - Sathyanarayana, Ubaradka G.

AU - Toyooka, Shinichi

AU - Padar, Asha

AU - Takahashi, Takashi

AU - Brambilla, Elizabeth

AU - Minna, John D.

AU - Gazdar, Adi F.

PY - 2003/7/1

Y1 - 2003/7/1

N2 - Purpose: We investigated the loss of expression of three laminin-5 (LN5)-encoding genes in lung cancer cell lines and elucidated the mechanism of inactivation of the genes in lung cancer cell lines and tumors. Experimental Design: We examined the expression of LN5-encoding genes by reverse transcription-PCR in 49 lung cancer cell lines. To elucidate the mechanism of gene silencing, we treated expression-negative cell lines (two for each gene) with a demethylating agent and examined the restoration of expression by reverse transcription-PCR. We dissected out the methylation patterns of CpG sites unique to the promoter regions of LN5-encoding genes by bisulfite genomic sequencing of expression-negative cell lines. We designed methylation-specific primers and validated the methylation status of the promoter regions in lung cancer cell lines using methylation-specific PCR. We further studied the methylation patterns of primary non-small cell lung cancer [NSCLC (n = 36)], small cell lung cancer [SCLC (n = 26)], and carcinoids (n = 24) tumors. Results: We observed frequent losses of expression in NSCLC (20-60%) and SCLC (65-86%) cell lines. Expression of one or more genes was lost in 90% of SCLC cell lines and 65% of NSCLC cell lines. Treatment of expression-negative cell lines with demethylating agent restored expression in all of the cases. Methylation of LN5-encoding genes was present more frequently in SCLC cell lines (60-80%) than in NSCLC cell lines (15-60%), and at least one gene was methylated in 95% of SCLC and 60% of NSCLC cell lines. The concordances between loss of expression and methylation in 40 lung cancer cell lines for the three genes (90-95%) were statistically significant. Methylation was more frequent in SCLC tumors (58-77%) than in NSCLC tumors (22-42%) and carcinoids (13-33%), and at least one gene was methylated in 92% of SCLC tumors, 47% of NSCLC tumors, and 33% of carcinoids. Conclusions: Our results demonstrate frequent epigenetic inactivation of LN5-encoding genes in lung cancers, and these findings are of biological interest and are potentially of clinical importance.

AB - Purpose: We investigated the loss of expression of three laminin-5 (LN5)-encoding genes in lung cancer cell lines and elucidated the mechanism of inactivation of the genes in lung cancer cell lines and tumors. Experimental Design: We examined the expression of LN5-encoding genes by reverse transcription-PCR in 49 lung cancer cell lines. To elucidate the mechanism of gene silencing, we treated expression-negative cell lines (two for each gene) with a demethylating agent and examined the restoration of expression by reverse transcription-PCR. We dissected out the methylation patterns of CpG sites unique to the promoter regions of LN5-encoding genes by bisulfite genomic sequencing of expression-negative cell lines. We designed methylation-specific primers and validated the methylation status of the promoter regions in lung cancer cell lines using methylation-specific PCR. We further studied the methylation patterns of primary non-small cell lung cancer [NSCLC (n = 36)], small cell lung cancer [SCLC (n = 26)], and carcinoids (n = 24) tumors. Results: We observed frequent losses of expression in NSCLC (20-60%) and SCLC (65-86%) cell lines. Expression of one or more genes was lost in 90% of SCLC cell lines and 65% of NSCLC cell lines. Treatment of expression-negative cell lines with demethylating agent restored expression in all of the cases. Methylation of LN5-encoding genes was present more frequently in SCLC cell lines (60-80%) than in NSCLC cell lines (15-60%), and at least one gene was methylated in 95% of SCLC and 60% of NSCLC cell lines. The concordances between loss of expression and methylation in 40 lung cancer cell lines for the three genes (90-95%) were statistically significant. Methylation was more frequent in SCLC tumors (58-77%) than in NSCLC tumors (22-42%) and carcinoids (13-33%), and at least one gene was methylated in 92% of SCLC tumors, 47% of NSCLC tumors, and 33% of carcinoids. Conclusions: Our results demonstrate frequent epigenetic inactivation of LN5-encoding genes in lung cancers, and these findings are of biological interest and are potentially of clinical importance.

UR - http://www.scopus.com/inward/record.url?scp=0037817329&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037817329&partnerID=8YFLogxK

M3 - Article

C2 - 12855645

AN - SCOPUS:0037817329

VL - 9

SP - 2665

EP - 2672

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 7

ER -