Tissue-based quantification of 8-hydroxy-2'-deoxyguanosine in human prostate biopsies using quantitative fluorescence imaging analysis.
OBJECTIVE
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.
Richardson
Tim
T
Department of Urology, University of Nebraska Medical Center and Omaha Veterans Affairs Hospital, Omaha, Nebraska 68114, USA. tarichardson@urologycenterpc.net
McCanse
Webb
W
Casale
George P
GP
Huang
Dali
D
Tian
Jennifer
J
Elkahwaji
Johny E
JE
Lele
Subodh
S
Hemstreet
George P
GP
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000368:Aged; D001706:Biopsy; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis; D006801:Humans; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D010865:Pilot Projects; D011467:Prostate / Q000737:chemistry* / Q000473:pathology
D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine