Original articleGenomic assessments of the frequent loss of heterozygosity region on 8p21.3∼p22 in head and neck squamous cell carcinoma
Introduction
Squamous cell carcinoma (SCC) of the head and neck is one of the most common cancers. In the United States, there are >31,000 new cases of head and neck or oral squamous cell carcinoma (HNSCC or OSCC) each year [1]. HNSCC will cause >8,000 deaths, killing ∼1 person per hour. Worldwide, the problem is much worse, with >270,000 new cases diagnosed each year [2]. In some parts of the world, including Melanesia, France, and the Indian subcontinent, HNSCC is a major health problem [3]. The overall 5-year survival rates for HNSCC have remained at ∼50%, considerably lower than for cervical cancer, Hodgkin's disease, or cancer of the brain, liver, testes, kidney, or skin (malignant melanoma) [2].
Tobacco, alcohol, and viral infections are common risk factors for HNSCC [4], [5], as are genetic polymorphisms on genes that metabolize carcinogens. Several chromosome regions have been identified as frequently altered in HNSCC, including 3p, 4q, 5q21∼q22, 8p21∼p23, 9p21∼p22, 11q13, 11q23, 13q, 14q, 17p, 18q, and 22q [6]. Significant improvement of functional mapping is needed to move forward research, diagnosis, and treatment of HNSCC.
Most human cancers are characterized by genetic instabilities [7]. Chromosomal aberrations include segments of allelic imbalance identifiable by loss of heterozygosity (LOH) at polymorphic loci, and may be used to implicate regions harboring tumor suppressor genes [8], [9], [10]. LOH patterns can be generated through allelotyping using polymorphic microsatellite markers. Only a modest number of microsatellite makers can be screened, however, because of the limited number of available microsatellite markers, the tedious and labor-intensive procedure, and the need for large amounts of DNA. Recent advances in high-density single-nucleotide polymorphism (SNP) array platforms provide unique opportunities for generating high-resolution LOH profiles. Our previous studies demonstrated the feasibility of using the 10K SNP mapping array (Affymetrix, Santa Clara, CA) for genome-wide LOH profiling [11], [12], [13].
In the present study, we used SNP array-based LOH profiling on a large panel of HNSCC cell lines to narrow the frequent LOH region at 8p to a region of ∼7 Mb, located at 8p21.3∼p22. Mitochondrial tumor suppressor gene 1 (MTUS1) is one of the candidate genes that reside in this genomic region. MTUS1 was initially identified as a potential tumor suppressor gene in pancreatic cancer [14]. Our recent study suggested that the reduction of MTUS1 expression may be associated with advanced oral tongue SCC [15]. Reduced expression of MTUS1 has been observed also in colon cancer, ovarian cancer, and pancreatic cancer [14], [16], [17]. In the present study, our expressional and sequence analyses on MTUS1 gene provide additional evidence in support of MTUS1 as a tumor suppressor for HNSCC.
Section snippets
Materials and methods
The SNP array assay was performed as described previously [11], [12], [13]. In brief, the genomic DNAs were isolated from cultured cell lines using a genomic DNA isolation kit (Qiagen, Valencia, CA). (For description of the 41 HNSCC and 4 reference cell lines, see Supplementary Table 1, available at http://www.elsevier.com.)
Labeling, hybridization, washing, and staining of the 10K SNP mapping array was performed according to the standard single primer GeneChip mapping assay protocol
Results and discussion
The SNP array-based LOH profiling was performed on 41 HNSCC cell lines. As reference, LOH profiles were also obtained for 2 immortalized normal oral keratinocyte, 1 normal oral keratinocyte primary culture (NHOK) and 1 immortalized skin keratinocyte (HaCaT). Although minimum allelic imbalance was observed for the normal keratinocytes, several frequent LOH regions were identified for HNSCC cases (Table 1). Chromosome arms 3p, 4p, 4q, 5q, 8p, 9p, 10p, 11q, and 17p exhibit frequent LOH, which is
Acknowledgments
This work was supported in part by National Institutes of Health Public Health Service (NIH-PHS) grants K22 DE014847, R03 DE016569, R03 CA114688 (to X.Z.), and R01 DE015970 (to D.T.W.). The 10K SNP mapping array hybridization and scanning were performed at the University of California Los Angeles (UCLA) DNA microarray facility. The sequence analyses performed at the University of Illinois at Chicago (UIC) Research Resource Center, DNA service facility. The primary NHOK cell and the 183 and 1483
References (35)
- et al.
Metabolizing enzyme genotype and risk for upper aerodigestive tract cancer
Oral Oncol
(2000) - et al.
Oral cancer: reviewing the present understanding of its molecular mechanism and exploring the future directions for its effective management
Oral Oncol
(2003) - et al.
Whole genome loss of heterozygosity profiling on oral squamous cell carcinoma by high-density single nucleotide polymorphic allele (SNP) array
Cancer Genet Cytogenet
(2004) - et al.
Progress in concurrent analysis of loss of heterozygosity and comparative genomic hybridization utilizing high density single nucleotide polymorphism arrays
Cancer Genet Cytogenet
(2005) - et al.
Global expression-based classification of lymph node metastasis and extracapsular spread of oral tongue squamous cell carcinoma
Neoplasia
(2006) - et al.
Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method
Methods
(2001) - et al.
Mutation analysis of the 8p22 candidate tumor suppressor gene ATIP/MTUS1 in hepatocellular carcinoma
Mol Cell Endocrinol
(2006) - et al.
Cytogenetic abnormalities in 106 oral squamous cell carcinomas
Cancer Genet Cytogenet
(2006) - et al.
Trans-inactivation of receptor tyrosine kinases by novel angiotensin II AT2 receptor-interacting protein, ATIP
J Biol Chem
(2004) - et al.
Triggering cell death: the crystal structure of Apo2L/TRAIL in a complex with death receptor 5
Mol Cell
(1999)
Structural organization and expression of human MTUS1, a candidate 8p22 tumor suppressor gene encoding a family of angiotensin II AT2 receptor-interacting proteins, ATIP
Gene
Cancer statistics, 2006
CA Cancer J Clin
Global cancer statistics, 2002
CA Cancer J Clin
Estimates of the worldwide incidence of 25 major cancers in 1990
Int J Cancer
Identification of risk groups for oral precancer and cancer and preventive measures
Clin Oral Investig
Advancement in characterization of genomic alterations for improved diagnosis, treatment and prognostics in cancer
Expert Rev Mol Diagn
FHIT as tumor suppressor: mechanisms and therapeutic opportunities
Cancer Biol Ther
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