Hypermethylated MAL gene - A silent marker of early colon tumorigenesis

Guro E. Lind, Terje Ahlquist, Matthias Kolberg, Marianne Berg, Mette Eknæs, Miguel A. Alonso, Anne Kallioniemi, Gunn I. Meling, Rolf I. Skotheim, Torleiv O. Rognum, Espen Thiis-Evensen, Ragnhild A. Lothe

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Background: Tumor-derived aberrantly methylated DNA might serve as diagnostic biomarkers for cancer, but so far, few such markers have been identified. The aim of the present study was to investigate the potential of the MAL (T-cell differentiation protein) gene as an early epigenetic diagnostic marker for colorectal tumors. Methods: Using methylation-specific polymerase chain reaction (MSP) the promoter methylation status of MAL was analyzed in 218 samples, including normal mucosa (n = 44), colorectal adenomas (n = 63), carcinomas (n = 65), and various cancer cell lines (n = 46). Direct bisulphite sequencing was performed to confirm the MSP results. MAL gene expression was investigated with real time quantitative analyses before and after epigenetic drug treatment. Immunohistochemical analysis of MAL was done using normal colon mucosa samples (n = 5) and a tissue microarray with 292 colorectal tumors. Results: Bisulphite sequencing revealed that the methylation was unequally distributed within the MAL promoter and by MSP analysis a region close to the transcription start point was shown to be hypermethylated in the majority of colorectal carcinomas (49/61, 80%) as well as in adenomas (45/63, 71%). In contrast, only a minority of the normal mucosa samples displayed hypermethylation (1/23, 4%). The hypermethylation of MAL was significantly associated with reduced or lost gene expression in in vitro models. Furthermore, removal of the methylation re-induced gene expression in colon cancer cell lines. Finally, MAL protein was expressed in epithelial cells of normal colon mucosa, but not in the malignant cells of the same type. Conclusion: Promoter hypermethylation of MAL was present in the vast majority of benign and malignant colorectal tumors, and only rarely in normal mucosa, which makes it suitable as a diagnostic marker for early colorectal tumorigenesis.

Original languageEnglish (US)
Article number13
JournalJournal of translational medicine
Volume6
DOIs
StatePublished - Mar 17 2008

Fingerprint

Methylation
Colon
Carcinogenesis
Mucous Membrane
Genes
Tumors
Colorectal Neoplasms
Gene expression
Gene Expression
Myelin and Lymphocyte-Associated Proteolipid Proteins
Epigenomics
Adenoma
Cells
Drug therapy
Cell Line
T-cells
Polymerase chain reaction
Transcription
Microarrays
Tumor Biomarkers

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Hypermethylated MAL gene - A silent marker of early colon tumorigenesis. / Lind, Guro E.; Ahlquist, Terje; Kolberg, Matthias; Berg, Marianne; Eknæs, Mette; Alonso, Miguel A.; Kallioniemi, Anne; Meling, Gunn I.; Skotheim, Rolf I.; Rognum, Torleiv O.; Thiis-Evensen, Espen; Lothe, Ragnhild A.

In: Journal of translational medicine, Vol. 6, 13, 17.03.2008.

Research output: Contribution to journalArticle

Lind, Guro E. ; Ahlquist, Terje ; Kolberg, Matthias ; Berg, Marianne ; Eknæs, Mette ; Alonso, Miguel A. ; Kallioniemi, Anne ; Meling, Gunn I. ; Skotheim, Rolf I. ; Rognum, Torleiv O. ; Thiis-Evensen, Espen ; Lothe, Ragnhild A. / Hypermethylated MAL gene - A silent marker of early colon tumorigenesis. In: Journal of translational medicine. 2008 ; Vol. 6.
@article{77e7f9750d3b488fb57e632ef89a0052,
title = "Hypermethylated MAL gene - A silent marker of early colon tumorigenesis",
abstract = "Background: Tumor-derived aberrantly methylated DNA might serve as diagnostic biomarkers for cancer, but so far, few such markers have been identified. The aim of the present study was to investigate the potential of the MAL (T-cell differentiation protein) gene as an early epigenetic diagnostic marker for colorectal tumors. Methods: Using methylation-specific polymerase chain reaction (MSP) the promoter methylation status of MAL was analyzed in 218 samples, including normal mucosa (n = 44), colorectal adenomas (n = 63), carcinomas (n = 65), and various cancer cell lines (n = 46). Direct bisulphite sequencing was performed to confirm the MSP results. MAL gene expression was investigated with real time quantitative analyses before and after epigenetic drug treatment. Immunohistochemical analysis of MAL was done using normal colon mucosa samples (n = 5) and a tissue microarray with 292 colorectal tumors. Results: Bisulphite sequencing revealed that the methylation was unequally distributed within the MAL promoter and by MSP analysis a region close to the transcription start point was shown to be hypermethylated in the majority of colorectal carcinomas (49/61, 80{\%}) as well as in adenomas (45/63, 71{\%}). In contrast, only a minority of the normal mucosa samples displayed hypermethylation (1/23, 4{\%}). The hypermethylation of MAL was significantly associated with reduced or lost gene expression in in vitro models. Furthermore, removal of the methylation re-induced gene expression in colon cancer cell lines. Finally, MAL protein was expressed in epithelial cells of normal colon mucosa, but not in the malignant cells of the same type. Conclusion: Promoter hypermethylation of MAL was present in the vast majority of benign and malignant colorectal tumors, and only rarely in normal mucosa, which makes it suitable as a diagnostic marker for early colorectal tumorigenesis.",
author = "Lind, {Guro E.} and Terje Ahlquist and Matthias Kolberg and Marianne Berg and Mette Ekn{\ae}s and Alonso, {Miguel A.} and Anne Kallioniemi and Meling, {Gunn I.} and Skotheim, {Rolf I.} and Rognum, {Torleiv O.} and Espen Thiis-Evensen and Lothe, {Ragnhild A.}",
year = "2008",
month = "3",
day = "17",
doi = "10.1186/1479-5876-6-13",
language = "English (US)",
volume = "6",
journal = "Journal of Translational Medicine",
issn = "1479-5876",
publisher = "BioMed Central",

}

TY - JOUR

T1 - Hypermethylated MAL gene - A silent marker of early colon tumorigenesis

AU - Lind, Guro E.

AU - Ahlquist, Terje

AU - Kolberg, Matthias

AU - Berg, Marianne

AU - Eknæs, Mette

AU - Alonso, Miguel A.

AU - Kallioniemi, Anne

AU - Meling, Gunn I.

AU - Skotheim, Rolf I.

AU - Rognum, Torleiv O.

AU - Thiis-Evensen, Espen

AU - Lothe, Ragnhild A.

PY - 2008/3/17

Y1 - 2008/3/17

N2 - Background: Tumor-derived aberrantly methylated DNA might serve as diagnostic biomarkers for cancer, but so far, few such markers have been identified. The aim of the present study was to investigate the potential of the MAL (T-cell differentiation protein) gene as an early epigenetic diagnostic marker for colorectal tumors. Methods: Using methylation-specific polymerase chain reaction (MSP) the promoter methylation status of MAL was analyzed in 218 samples, including normal mucosa (n = 44), colorectal adenomas (n = 63), carcinomas (n = 65), and various cancer cell lines (n = 46). Direct bisulphite sequencing was performed to confirm the MSP results. MAL gene expression was investigated with real time quantitative analyses before and after epigenetic drug treatment. Immunohistochemical analysis of MAL was done using normal colon mucosa samples (n = 5) and a tissue microarray with 292 colorectal tumors. Results: Bisulphite sequencing revealed that the methylation was unequally distributed within the MAL promoter and by MSP analysis a region close to the transcription start point was shown to be hypermethylated in the majority of colorectal carcinomas (49/61, 80%) as well as in adenomas (45/63, 71%). In contrast, only a minority of the normal mucosa samples displayed hypermethylation (1/23, 4%). The hypermethylation of MAL was significantly associated with reduced or lost gene expression in in vitro models. Furthermore, removal of the methylation re-induced gene expression in colon cancer cell lines. Finally, MAL protein was expressed in epithelial cells of normal colon mucosa, but not in the malignant cells of the same type. Conclusion: Promoter hypermethylation of MAL was present in the vast majority of benign and malignant colorectal tumors, and only rarely in normal mucosa, which makes it suitable as a diagnostic marker for early colorectal tumorigenesis.

AB - Background: Tumor-derived aberrantly methylated DNA might serve as diagnostic biomarkers for cancer, but so far, few such markers have been identified. The aim of the present study was to investigate the potential of the MAL (T-cell differentiation protein) gene as an early epigenetic diagnostic marker for colorectal tumors. Methods: Using methylation-specific polymerase chain reaction (MSP) the promoter methylation status of MAL was analyzed in 218 samples, including normal mucosa (n = 44), colorectal adenomas (n = 63), carcinomas (n = 65), and various cancer cell lines (n = 46). Direct bisulphite sequencing was performed to confirm the MSP results. MAL gene expression was investigated with real time quantitative analyses before and after epigenetic drug treatment. Immunohistochemical analysis of MAL was done using normal colon mucosa samples (n = 5) and a tissue microarray with 292 colorectal tumors. Results: Bisulphite sequencing revealed that the methylation was unequally distributed within the MAL promoter and by MSP analysis a region close to the transcription start point was shown to be hypermethylated in the majority of colorectal carcinomas (49/61, 80%) as well as in adenomas (45/63, 71%). In contrast, only a minority of the normal mucosa samples displayed hypermethylation (1/23, 4%). The hypermethylation of MAL was significantly associated with reduced or lost gene expression in in vitro models. Furthermore, removal of the methylation re-induced gene expression in colon cancer cell lines. Finally, MAL protein was expressed in epithelial cells of normal colon mucosa, but not in the malignant cells of the same type. Conclusion: Promoter hypermethylation of MAL was present in the vast majority of benign and malignant colorectal tumors, and only rarely in normal mucosa, which makes it suitable as a diagnostic marker for early colorectal tumorigenesis.

UR - http://www.scopus.com/inward/record.url?scp=42349095589&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=42349095589&partnerID=8YFLogxK

U2 - 10.1186/1479-5876-6-13

DO - 10.1186/1479-5876-6-13

M3 - Article

C2 - 18346269

AN - SCOPUS:42349095589

VL - 6

JO - Journal of Translational Medicine

JF - Journal of Translational Medicine

SN - 1479-5876

M1 - 13

ER -