N-Myc Downstream-Regulated Gene 4 ( NDRG4 ): A Candidate Tumor Suppressor Gene and Potential Biomarker for Colorectal Cancer
- 1 July 2009
- journal article
- Published by Oxford University Press (OUP) in JNCI Journal of the National Cancer Institute
- Vol. 101 (13), 916-927
- https://doi.org/10.1093/jnci/djp131
Abstract
Identification of hypermethylated tumor suppressor genes in body fluids is an appealing strategy for the noninvasive detection of colorectal cancer. Here we examined the role of N-Myc downstream-regulated gene 4 (NDRG4) as a novel tumor suppressor and biomarker in colorectal cancer. NDRG4 promoter methylation was analyzed in human colorectal cancer cell lines, colorectal tissue, and noncancerous colon mucosa by using methylation-specific polymerase chain reaction (PCR) and bisulfite sequencing. NDRG4 mRNA and protein expression were studied using real-time-PCR and immunohistochemistry, respectively. Tumor suppressor functions of NDRG4 were examined by colony formation, cell proliferation, and migration and invasion assays in colorectal cancer cell lines that were stably transfected with an NDRG4 expression construct. Quantitative methylation-specific PCR was used to examine the utility of NDRG4 promoter methylation as a biomarker in fecal DNA from 75 colorectal cancer patients and 75 control subjects. All P values are two-sided. The prevalence of NDRG4 promoter methylation in two independent series of colorectal cancers was 86% (71/83) and 70% (128/184) compared with 4% (2/48) in noncancerous colon mucosa (P < .001). NDRG4 mRNA and protein expression were decreased in colorectal cancer tissue compared with noncancerous colon mucosa. NDRG4 overexpression in colorectal cancer cell lines suppressed colony formation (P = .014), cell proliferation (P < .001), and invasion (P < .001). NDRG4 promoter methylation analysis in fecal DNA from a training set of colorectal cancer patients and control subjects yielded a sensitivity of 61% (95% confidence interval [CI] = 43% to 79%) and a specificity of 93% (95% CI = 90% to 97%). An independent test set of colorectal cancer patients and control subjects yielded a sensitivity of 53% (95% CI = 39% to 67%) and a specificity of 100% (95% CI = 86% to 100%). NDRG4 is a candidate tumor suppressor gene in colorectal cancer whose expression is frequently inactivated by promoter methylation. NDRG4 promoter methylation is a potential biomarker for the noninvasive detection of colorectal cancer in stool samples.Keywords
This publication has 41 references indexed in Scilit:
- Adjusting for Covariates in Studies of Diagnostic, Screening, or Prognostic Markers: An Old Concept in a New SettingAmerican Journal of Epidemiology, 2008
- Novel Blood-Based, Five-Gene Biomarker Set for the Detection of Colorectal CancerClinical Cancer Research, 2008
- Integrated analysis of chromosomal, microsatellite and epigenetic instability in colorectal cancer identifies specific associations between promoter methylation of pivotal tumour suppressor and DNA repair genes and specific chromosomal alterationsCarcinogenesis: Integrative Cancer Research, 2007
- A Novel Method to Capture Methylated Human DNA from Stool: Implications for Colorectal Cancer ScreeningClinical Chemistry, 2007
- Promoter methylation, mutation, and genomic deletion are involved in the decreased NDRG2 expression levels in several cancer cell linesBiochemical and Biophysical Research Communications, 2007
- The Consensus Coding Sequences of Human Breast and Colorectal CancersScience, 2006
- Quantitative Analysis of Plasma DNA in Colorectal Cancer PatientsAnnals of the New York Academy of Sciences, 2006
- CpG island methylator phenotype underlies sporadic microsatellite instability and is tightly associated with BRAF mutation in colorectal cancerNature Genetics, 2006
- Angiogenesis gene expression profiling in xenograft models to study cellular interactionsExperimental Cell Research, 2004
- Loss of heterozygosity on chromosome 16 in hepatocellular carcinomaJournal of Gastroenterology and Hepatology, 1992