Stability of phosphoprotein as a biological marker of tumor signaling

Amanda F. Baker, Tomislav Dragovich, Nathan T. Ihle, Ryan Williams, Cecilia Fenoglio-Preiser, Garth Powis

Research output: Contribution to journalArticlepeer-review

137 Scopus citations


Purpose: The purpose of the study was to evaluate the stability of phosphoprotein as a marker of signaling activity in human tumors using clinical samples and xenografts. Experimental Design: The expression of phospho-Ser 473-Akt (p-Akt) was assessed by immunohistochemistry in paraffin-embedded samples from patients enrolled in a Southwest Oncology Group clinical trial of gastroesophageal junction tumors and by immunohistochemistry and Western blotting in human colon tumor xenografts at various times after removal from the animal. Results: Clinical samples had evaluable p-Akt staining only when obtained as biopsies (9 of 13) and no staining was observed in tumors obtained as surgically resected samples (0 of 15). In HT-29 colon cancer xenografts, p-Akt staining was present in fresh sample but not in tissue that had been allowed to stand for 30 minutes at room temperature. Western blotting of HT-29 tumor xenografts at room temperature showed a slow decrease in total Akt with a half-life of 180 minutes and a rapid decrease in p-Akt with a half-life of 20 minutes. Conclusions: Caution should be used when using phosphoprotein levels in human tumor specimens to measure intrinsic signaling activity or drug effects because of the potential for rapid dephosphorylation. Rapid processing of biopsies is essential and postoperative surgical samples may be of limited value because of the time to fixation.

Original languageEnglish (US)
Pages (from-to)4338-4340
Number of pages3
JournalClinical Cancer Research
Issue number12
StatePublished - Jun 15 2005

ASJC Scopus subject areas

  • Oncology
  • Cancer Research


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