Tissue-based Quantification of 8-Hydroxy-2′-Deoxyguanosine in Human Prostate Biopsies Using Quantitative Fluorescence Imaging Analysis

Tim Richardson, Webb McCanse, George P. Casale, Dali Huang, Jennifer Tian, Johny E. Elkahwaji, Subodh Lele, George P. Hemstreet

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Objectives: To quantify 8-hydroxy-2′-deoxyguanosine (8-OHdG) in prostate stromal and acini tissue compartments from benign and cancer-containing prostate specimens using a new quantitative fluorescence imaging analysis protocol. Methods: Prostate biopsy specimens from 20 age-matched benign (control) and cancer-containing tissue sections were used to quantify 8-OHdG. 8-OHdG was quantified within individual acini nuclei and the surrounding stroma nuclei. Paraffin sections were treated with RNAse and protease to expose the nuclear chromatin, reacted with anti-8-OHdG mouse monoclonal antibody bound to saturation and detected with secondary goat anti-mouse IgG labeled with Alex Fluor 488, and quantified with a calibrated quantitative fluorescence imaging analysis system. The results were analyzed using a paired Student's t test. Results: 8-OHdG was successfully quantified within individual cellular compartments without the need for laser tissue dissection, using the mean pixel intensity of fluorescent-labeled 8-OHdG. Matched-pair analysis of the global expression of 8-OHdG, as well as the acini and stroma individually, revealed no difference between the cancerous and control prostatic tissue. All patients with prostate cancer and those with benign findings had significantly greater 8-OHdG within the acini compared with the surrounding stoma (P < .05). Conclusions: A protocol to quantify 8-OHdG in paraffin-embedded human prostatic tissue was successfully developed. 8-OHdG was not significantly elevated in the acini or stroma of cancer-containing prostatic tissue compared with age-matched benign prostatic tissue. Although 8-OHdG was significantly elevated in the acini nuclei compared with the surrounding stroma nuclei in both cancer-containing and benign prostatic tissue, it, by itself, was not a strong biomarker for prostate cancer risk assessment.

Original languageEnglish (US)
Pages (from-to)1174-1179
Number of pages6
JournalUrology
Volume74
Issue number5
DOIs
StatePublished - Nov 2009

ASJC Scopus subject areas

  • Urology

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