The APC gene product in normal and tumor cells

Kelly J. Smith, Karen A. Johnson, Tracy M. Bryan, David E. Hill, Sanford Markowitz, James K V Willson, Christos Paraskeva, Gloria M Petersen, Stanley R. Hamilton, Bert Vogelstein, Kenneth W. Kinzler

Research output: Contribution to journalArticle

413 Citations (Scopus)

Abstract

The APC gene has been found to be mutated during the development of sporadic colorectal tumors as well as in the germ line of familial adenomatous polyposis patients. To facilitate the characterization of both normal and mutant APC protein, a series of monoclonal and polyclonal antibodies specific for the APC protein was produced. When lymphoblastoid cell lines derived from seven familial adenomatous polyposis patients with known mutations were analyzed by Western blot, an ≈300-kDa protein corresponding to the predicted size of full-length APC was detected in all 7 cell lines. In addition, truncated APC proteins corresponding to the product of the known mutated alleles could be detected in 4 of the 7 lines. Similar analysis of 23 colon carcinoma and 9 adenoma cell lines revealed truncated proteins in 24 (75%) of the cell lines. Moreover, 26 (81%) of the colon tumor lines were totally devoid of the normal, full-length protein. In contrast, Western blot analysis of 40 cell lines derived from sporadic tumors of other organs detected only full-length APC. Immunohistochemical analysis of APC in normal colonic mucosa revealed cytoplasmic staining with more intense staining in the basolateral margins of the epithelial cell. This staining was markedly increased in the upper portions of the crypts, suggesting an increased level of expression with maturation. These studies provide some initial clues to the function of the cytoplasmic protein APC and demonstrate the feasibility of identifying APC mutations by direct analysis of the APC protein.

Original languageEnglish (US)
Pages (from-to)2846-2850
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number7
StatePublished - Apr 1 1993
Externally publishedYes

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APC Genes
Cell Line
Neoplasms
Proteins
Adenomatous Polyposis Coli
Staining and Labeling
Colon
Western Blotting
Mutation
Mutant Proteins
Germ Cells
Adenoma
Colorectal Neoplasms
Mucous Membrane
Epithelial Cells
Alleles
Monoclonal Antibodies
Carcinoma

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Smith, K. J., Johnson, K. A., Bryan, T. M., Hill, D. E., Markowitz, S., Willson, J. K. V., ... Kinzler, K. W. (1993). The APC gene product in normal and tumor cells. Proceedings of the National Academy of Sciences of the United States of America, 90(7), 2846-2850.

The APC gene product in normal and tumor cells. / Smith, Kelly J.; Johnson, Karen A.; Bryan, Tracy M.; Hill, David E.; Markowitz, Sanford; Willson, James K V; Paraskeva, Christos; Petersen, Gloria M; Hamilton, Stanley R.; Vogelstein, Bert; Kinzler, Kenneth W.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 7, 01.04.1993, p. 2846-2850.

Research output: Contribution to journalArticle

Smith, KJ, Johnson, KA, Bryan, TM, Hill, DE, Markowitz, S, Willson, JKV, Paraskeva, C, Petersen, GM, Hamilton, SR, Vogelstein, B & Kinzler, KW 1993, 'The APC gene product in normal and tumor cells', Proceedings of the National Academy of Sciences of the United States of America, vol. 90, no. 7, pp. 2846-2850.
Smith KJ, Johnson KA, Bryan TM, Hill DE, Markowitz S, Willson JKV et al. The APC gene product in normal and tumor cells. Proceedings of the National Academy of Sciences of the United States of America. 1993 Apr 1;90(7):2846-2850.
Smith, Kelly J. ; Johnson, Karen A. ; Bryan, Tracy M. ; Hill, David E. ; Markowitz, Sanford ; Willson, James K V ; Paraskeva, Christos ; Petersen, Gloria M ; Hamilton, Stanley R. ; Vogelstein, Bert ; Kinzler, Kenneth W. / The APC gene product in normal and tumor cells. In: Proceedings of the National Academy of Sciences of the United States of America. 1993 ; Vol. 90, No. 7. pp. 2846-2850.
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