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Expression of five selected human mismatch repair genes simultaneously detected in normal and cancer cell lines by a nonradioactive multiplex reverse transcription-polymerase chain reaction.

Publication ,  Journal Article
Wei, Q; Guan, Y; Cheng, L; Radinsky, R; Bar-Eli, M; Tsan, R; Li, L; Legerski, RJ
Published in: Pathobiology
1997

Abnormalities in at least 1 of 5 mismatch repair (MMR) genes (hMSH2, hMLH1, hPMS1, hPMS2 and GTBP/hMSH6) are found in hereditary nonpolyposis colon cancer and sporadic colon cancers. We used a single-reaction multiplex reverse transcription (RT)-polymerase chain reaction (PCR), with the beta-actin gene as an internal control, to simultaneously evaluate expression of these 5 known human MMR genes in normal and tumor cell lines with known or uncharacterized mutations in MMR genes. The relative quantitation of the transcripts is demonstrated by controlling the number of PCR cycles and titrating cDNA with a dose-curve. The 13 normal cell lines tested were derived from normal lymphocytes, skin, thymus, breast, lung, colon, liver and kidney. The 26 cancer cell lines were derived from melanoma and cancers of the brain, breast, lung, colon, pancreas and prostate. All 5 MMR genes were ubiquitously expressed in all normal cell lines tested, suggesting their housekeeping roles. Aberrant MMR gene expression was only observed in the colon cancer cell lines. Two previously uncharacterized colon cancer cell lines did not express hMLH1. These data suggest that this nonradioactive multiplex RT-PCR assay for MMR gene expression may be useful for fast screening for genetic alterations that may affect gene expression and so may aid molecular analysis of MMR-related colon cancer.

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Published In

Pathobiology

DOI

ISSN

1015-2008

Publication Date

1997

Volume

65

Issue

6

Start / End Page

293 / 300

Location

Switzerland

Related Subject Headings

  • Tumor Cells, Cultured
  • Transcription, Genetic
  • RNA
  • Proto-Oncogene Proteins
  • Polymerase Chain Reaction
  • Oncology & Carcinogenesis
  • Nuclear Proteins
  • Neoplasm Proteins
  • Mutation
  • MutS Homolog 2 Protein
 

Citation

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Wei, Q., Guan, Y., Cheng, L., Radinsky, R., Bar-Eli, M., Tsan, R., … Legerski, R. J. (1997). Expression of five selected human mismatch repair genes simultaneously detected in normal and cancer cell lines by a nonradioactive multiplex reverse transcription-polymerase chain reaction. Pathobiology, 65(6), 293–300. https://doi.org/10.1159/000164141
Wei, Q., Y. Guan, L. Cheng, R. Radinsky, M. Bar-Eli, R. Tsan, L. Li, and R. J. Legerski. “Expression of five selected human mismatch repair genes simultaneously detected in normal and cancer cell lines by a nonradioactive multiplex reverse transcription-polymerase chain reaction.Pathobiology 65, no. 6 (1997): 293–300. https://doi.org/10.1159/000164141.
Journal cover image

Published In

Pathobiology

DOI

ISSN

1015-2008

Publication Date

1997

Volume

65

Issue

6

Start / End Page

293 / 300

Location

Switzerland

Related Subject Headings

  • Tumor Cells, Cultured
  • Transcription, Genetic
  • RNA
  • Proto-Oncogene Proteins
  • Polymerase Chain Reaction
  • Oncology & Carcinogenesis
  • Nuclear Proteins
  • Neoplasm Proteins
  • Mutation
  • MutS Homolog 2 Protein