Gastroenterology

Gastroenterology

Volume 109, Issue 4, October 1995, Pages 1368-1374
Gastroenterology

In vitro transcription/translation assay for the screening of hMLH1 and hMSH2 mutations in familial colon cancer

https://doi.org/10.1016/0016-5085(95)90600-2Get rights and content

Abstract

Background & Aims: Hereditary nonpolyposis colorectal cancer (HNPCC) has been linked recently to a defect in repairing mismatched nucleotides in DNA. The aim of this study was to screen for germline mutations that result in prematurely truncated proteins in two of the mismatch repair genes identified at this time, hMLH1 and hMSH2, in a consecutive series of patients belonging to familial aggregations of colorectal cancer. Methods: Nineteen individuals with colorectal cancer from 19 families were consecutively referred because of a strong positive family history of colorectal cancer. Premature truncation mutations in hMLH1 and hMSH2 were sought from lymphocyte RNA by using an in vitro transcription/translation (IVTT) assay. Results: Protein truncating mutations in the hMLH1 or hMSH2 genes were found in 50% of families with HNPCC (6 of 12) but were not observed in any of the remaining familial aggregations that did not fulfill the standard criteria for HNPCC. In some of the IVTT-positive samples, the mutations were characterized by genomic sequencing. Conclusions: IVTT may be a practical method to accomplish primary screening of germline mutations in DNA mismatch repair genes in HNPCC; however, a broader approach is necessary to obtain a more complete picture of the mutational spectrum in HNPCC and other familial aggregations of colorectal cancer.

References (33)

  • E Lovett

    Family studies in cancer of the colon and rectum

    Br J Surg

    (1976)
  • RS Houlston et al.

    Screening and genetic counselling for relatives of patients with colorectal cancer in a family cancer clinic

    BMJ

    (1990)
  • CS Fuchs et al.

    A prospective study of family history and the risk of colorectal cancer

    N Engl J Med

    (1994)
  • JE Bailey-Wilson et al.

    Segregation analysis of hereditary non-polyposis colorectal cancer

    Genet Epidemiol

    (1986)
  • J-P Mecklin et al.

    Screening for colorectal carcinoma in cancer family syndrome kindreds

    Scand J Gastroenterol

    (1987)
  • M Ponz de Leon et al.

    Familial aggregation of tumors in the three year experience of a population-based colorectal cancer registry

    Cancer Res

    (1989)
  • Cited by (62)

    • Germline characterization of early-aged onset of hereditary non-polyposis colorectal cancer

      2001, Journal of Pediatrics
      Citation Excerpt :

      PCR amplifications were performed by using primers for hMLH1 complementary DNA to amplify the gene into 2 overlapping segments. IVTT was performed, and the product was electrophoresed onto a 12.5% sodium dodecylsulfate–polyacrylamide gel as previously described.19 The gel was fixed and prepared for fluorography with Entensity (New England Nuclear, Beverly, Mass).

    • An American founder mutation in MLH1

      2012, International Journal of Cancer
    View all citing articles on Scopus

    Supported by National Institutes of Health grant CA46592-06 from the University of Michigan Comprehensive Cancer Center, U.S. Public Health Service grant CA39233 from the Research Service of the Department of Veterans Affairs, and the Johnson Family Fund for Familial Colon Cancer Research.

    1

    Dr. Koi's current affiliation is: Laboratory of Molecular Carcinogenesis, National Institute for Environmental Health Sciences, Research Triangle Park, North Carolina.

    View full text