Assessment of apoptosis by immunohistochemical markers compared to cellular morphology in ex vivo-stressed colonic mucosa

Hana Holubec, Claire M. Payne, Harris Bernstein, Katerina Dvorakova, Carol Bernstein, Caroline N. Waltmire, James A Warneke, Harinder Garewal

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Apoptosis competence is central to the prevention of cancer. Frequency of apoptotic cells, after a sample of colonic tissue is stressed, can be used to gauge apoptosis competence and, thus, possible susceptibility to colon cancer. The gold standard for assessment of apoptosis is morphological evaluation, but this requires an experienced microscopist. Easier-to-use immunohistochemical markers of apoptosis, applicable in archived paraffin-embedded tissue, have been commercially developed. Potentially useful apoptosis markers include cleaved cytokeratin-18 (c-CK18), cleaved caspase-3 (c-cas-3), cleaved lamin A (c-lam-A), phosphorylated histone H2AX (γH2AX), cleaved poly(ADP ribose) polymerase (c-PARP), and translocation of apoptosis-inducing factor (AIF). When tissue samples from freshly resected colon segments were challenged ex vivo with the bile acid deoxycholate, ∼50% of goblet cells became apoptotic by morphologic criteria. This high level of morphologic apoptosis allowed quantitative comparison with the usefulness and specificity of immunohistochemical markers of apoptosis. The antibody to C-CK18 was almost as useful and about as specific as morphology for identifying apoptotic colonic epithelial cells. Antibodies to c-cas-3, c-lam-A, and γH2AX, though specific for apoptotic cells, were less useful. The antibody to c-PARP, though specific for apoptotic cells, had low usefulness, and the antibody to AIF was relatively nonspecific, under our conditions.

Original languageEnglish (US)
Pages (from-to)229-235
Number of pages7
JournalJournal of Histochemistry and Cytochemistry
Volume53
Issue number2
DOIs
StatePublished - Feb 2005

Fingerprint

Mucous Membrane
Apoptosis
Apoptosis Inducing Factor
Lamin Type A
Poly(ADP-ribose) Polymerases
Antibodies
Caspase 3
Mental Competency
Keratin-18
Deoxycholic Acid
Goblet Cells
Bile Acids and Salts
Paraffin
Histones
Colonic Neoplasms
Colon
Epithelial Cells
Neoplasms

Keywords

  • Apoptosis
  • Bile acid
  • Caspase-3
  • Colonic mucosa
  • Cytokeratin-18
  • Deoxycholate
  • Histone H2AX
  • Lamin A

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Assessment of apoptosis by immunohistochemical markers compared to cellular morphology in ex vivo-stressed colonic mucosa. / Holubec, Hana; Payne, Claire M.; Bernstein, Harris; Dvorakova, Katerina; Bernstein, Carol; Waltmire, Caroline N.; Warneke, James A; Garewal, Harinder.

In: Journal of Histochemistry and Cytochemistry, Vol. 53, No. 2, 02.2005, p. 229-235.

Research output: Contribution to journalArticle

Holubec, Hana ; Payne, Claire M. ; Bernstein, Harris ; Dvorakova, Katerina ; Bernstein, Carol ; Waltmire, Caroline N. ; Warneke, James A ; Garewal, Harinder. / Assessment of apoptosis by immunohistochemical markers compared to cellular morphology in ex vivo-stressed colonic mucosa. In: Journal of Histochemistry and Cytochemistry. 2005 ; Vol. 53, No. 2. pp. 229-235.
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