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PLoS One. 2014 Jul 1;9(7):e101010. doi: 10.1371/journal.pone.0101010. eCollection 2014.

Telomerase variant A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal carcinomas.

Author information

  • 1Thoracic Surgery Section, Thoracic and GI Oncology Branch; National Cancer Institute, Bethesda, Maryland, United States of America.
  • 2National Heart, Lung, and Blood Institute, Bethesda, Maryland, United States of America.
  • 3Departments of Pathology and Oncology, Johns Hopkins University of Medicine, Baltimore, Maryland, United States of America.
  • 4Laboratory of Cellular Oncology, National Cancer Institute, Bethesda, Maryland, United States of America.
  • 5Laboratory of Experimental Carcinogenesis, National Cancer Institute, Bethesda, Maryland, United States of America.
  • 6Section of Cancer Genomics, National Cancer Institute, Bethesda, Maryland, United States of America.
  • 7Comparative Molecular Cytogenetics Core Facility, National Cancer Institute, Frederick, Maryland, United States of America.
  • 8Department of Thoracic Surgery, Weill Cornell Medical Center, New York, New York, United States of America.
  • 9Department of Surgery, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
  • 10Section of Thoracic Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, United States of America.
  • 11Laboratory of Human Carcinogenesis, National Cancer Institute, Bethesda, Maryland, United States of America.

Abstract

BACKGROUND:

Although implicated in the pathogenesis of several chronic inflammatory disorders and hematologic malignancies, telomerase mutations have not been thoroughly characterized in human cancers. The present study was performed to examine the frequency and potential clinical relevance of telomerase mutations in esophageal carcinomas.

METHODS:

Sequencing techniques were used to evaluate mutational status of telomerase reverse transcriptase (TERT) and telomerase RNA component (TERC) in neoplastic and adjacent normal mucosa from 143 esophageal cancer (EsC) patients. MTS, flow cytometry, time lapse microscopy, and murine xenograft techniques were used to assess proliferation, apoptosis, chemotaxis, and tumorigenicity of EsC cells expressing either wtTERT or TERT variants. Immunoprecipitation, immunoblot, immunofluorescence, promoter-reporter and qRT-PCR techniques were used to evaluate interactions of TERT and several TERT variants with BRG-1 and β-catenin, and to assess expression of cytoskeletal proteins, and cell signaling. Fluorescence in-situ hybridization and spectral karyotyping techniques were used to examine telomere length and chromosomal stability.

RESULTS:

Sequencing analysis revealed one deletion involving TERC (TERC del 341-360), and two non-synonymous TERT variants [A279T (2 homozygous, 9 heterozygous); A1062T (4 heterozygous)]. The minor allele frequency of the A279T variant was five-fold higher in EsC patients compared to healthy blood donors (p<0.01). Relative to wtTERT, A279T decreased telomere length, destabilized TERT-BRG-1-β-catenin complex, markedly depleted β-catenin, and down-regulated canonical Wnt signaling in cancer cells; these phenomena coincided with decreased proliferation, depletion of additional cytoskeletal proteins, impaired chemotaxis, increased chemosensitivity, and significantly decreased tumorigenicity of EsC cells. A279T expression significantly increased chromosomal aberrations in mouse embryonic fibroblasts (MEFs) following Zeocin™ exposure, as well as Li Fraumeni fibroblasts in the absence of pharmacologically-induced DNA damage.

CONCLUSIONS:

A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal cancer cells. These findings warrant further analysis of A279T expression in esophageal cancers and premalignant esophageal lesions.

PMID:
24983628
[PubMed - in process]
PMCID:
PMC4077737
Free PMC Article
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