Monochromosome transfer and microarray analysis identify a critical tumor-suppressive region mapping to chromosome 13q14 and THSD1 in esophageal carcinoma

Mol Cancer Res. 2008 Apr;6(4):592-603. doi: 10.1158/1541-7786.MCR-07-0154.


Loss of chromosome 13q regions in esophageal squamous cell carcinoma (ESCC) is a frequent event. Monochromosome transfer approaches provide direct functional evidence for tumor suppression by chromosome 13 in SLMT-1, an ESCC cell line, and identify critical regions at 13q12.3, 13q14.11, and 13q14.3. Differential gene expression profiles of three tumor-suppressing microcell hybrids (MCH) and their tumorigenic parental SLMT-1 cell line were revealed by competitive hybridization using 19k cDNA oligonucleotide microarrays. Nine candidate 13q14 tumor-suppressor genes (TSG), including RB1, showed down-regulation in SLMT-1, compared with NE1, an immortalized normal esophageal epithelial cell line; their average gene expression was restored in MCHs compared with SLMT-1. Reverse transcription-PCR validated gene expression levels in MCHs and a panel of ESCC cell lines. Results suggest that the tumor-suppressing effect is not attributed to RB1, but instead likely involves thrombospondin type I domain-containing 1 (THSD1), a novel candidate TSG mapping to 13q14. Quantitative reverse transcription-PCR detected down-regulation of THSD1 expression in 100% of ESCC and other cancer cell lines. Mechanisms for THSD1 silencing in ESCC involved loss of heterozygosity and promoter hypermethylation, as analyzed by methylation-specific PCR and clonal bisulfite sequencing. Transfection of wild-type THSD1 into SLMT-1 resulted in significant reduction of colony-forming ability, hence providing functional evidence for its growth-suppressive activity. These findings suggest that THSD1 is a good candidate TSG.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Cell Line, Transformed
  • Cell Line, Tumor
  • Chromosome Mapping*
  • Chromosome Segregation
  • Chromosomes, Human, Pair 13 / genetics*
  • DNA Methylation / drug effects
  • Deoxycytidine / pharmacology
  • Epithelial Cells / drug effects
  • Epithelial Cells / pathology
  • Esophageal Neoplasms / genetics*
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic / drug effects
  • Genes, Tumor Suppressor*
  • Genome, Human / genetics
  • Humans
  • Hydroxamic Acids / pharmacology
  • In Situ Hybridization, Fluorescence
  • Microarray Analysis*
  • Microsatellite Repeats / genetics
  • Promoter Regions, Genetic / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Thrombospondins / genetics*
  • Thrombospondins / metabolism
  • Transfection
  • Tumor Stem Cell Assay


  • Hydroxamic Acids
  • THSD1 protein, human
  • Thrombospondins
  • Deoxycytidine
  • trichostatin A