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Digestive Diseases and Sciences

, Volume 44, Issue 3, pp 553–559 | Cite as

Reduction in hMSH2 mRNA Levels by Premature Translation Termination (Implications for Mutation Screening in Hereditary Nonpolyposis Colorectal Cancer)

  • Xuede Lin
  • Jai H. Choi
  • Patrick Lynch
  • Lixuan Xi
  • Elsie Wu
  • Marsha L. Frazier
Article

Abstract

We have used single-strand conformationalpolymorphism (SSCP) and heteroduplex analysis to examineDNA from 50 colorectal carcinoma patients coming fromfamilies meeting the Amsterdam criteria for hereditary nonpolyposis colorectal cancer (HNPCC) orhaving developed colorectal carcinoma at age 45 oryounger. We identified mutations in 12 of thesepatients, with seven of these being novel mutations. Weexamined four of the truncating mutations using in vitrotranscription and translation (IVTT) assays anddetermined that the mutation causing an in-framedeletion of exon 5 could easily be detected by the IVTTassay, but the three mutations resulting in prematuretranslation termination were not detected because thesteady-state levels of the mutant allele transcripts aretoo low. Our findings suggest that some but not all mutant hMSH2 alleles have significantlylower steady-state mRNA levels than the normal allele.Under ideal circumstances, where lymphoblastoid celllines are available for RNA extraction, IVTT may be useful for detecting truncating mutations.However, our data suggest that caution should be takenin using IVTT in routine screening of clinical samplesfor truncating HNPCC mutations, as many mutations may go undetected.

HEREDITARY NONPOLYPOSIS COLORECTAL CANCER HMSH2 MUTATION PROTEIN TRUNCATION HETERODUPLEX ANALYSIS SINGLE-STRAND CONFORMATIONAL POLYMORPHISMANALYSIS 

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Copyright information

© Plenum Publishing Corporation 1999

Authors and Affiliations

  • Xuede Lin
  • Jai H. Choi
  • Patrick Lynch
  • Lixuan Xi
  • Elsie Wu
  • Marsha L. Frazier

There are no affiliations available

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