Production of 13-hydroxyoctadecadienoic acid (13-HODE) by prostate tumors and cell lines.
The major lipoxygenation product derived from linoleic acid, 13-(S)-hydroxyoctadecadienoic acid (13-HODE), has been shown to be involved in cell proliferation and differentiation in a number of systems. Rapid detection of picogram amounts of this bioactive lipid in biological samples, however, has been hindered due to lack of immunological reagents. In the current report, we have used a polyclonal antibody specific for 13-(S)-HODE to detect this bioactive lipid for the first time in human prostate adenocarcinoma specimens (PCa) and the prostate cancer cell lines LNCaP and PC-3 by enzyme immunoassay. In addition, we have verified-the quantitation of 13-HODE by chiral-phase HPLC and examined the levels of lipoxygenase expression by Western, Northern, and RT-PCR analysis. Immunohistochemically detectable 13-HODE was observed in human PCa, whereas adjacent normal tissue showed no immunoreactivity. The presence of 15-lipoxygenase was evident by Western and RT-PCR analysis in both LNCaP and PC-3 cells, while Northern blot analysis showed the presence of 15-lipoxygenase message in LNCaP cells but failed to detect any 15-lipoxygenase message in PC-3 cells. In contrast, quantitation of 13-HODE by enzyme immunoassay and chiral-phase HPLC showed significant levels of the compound in PC-3 cells but minimal enzymatically produced 13-HODE in LNCaP cells. These data provide a link between linoleic acid metabolism and the development or progression of prostate cancer.
Spindler
S A
SA
Oxford Biomedical Research, Inc., Rochester Hills, Michigan 48309, USA.
Sarkar
F H
FH
Sakr
W A
WA
Blackburn
M L
ML
Bull
A W
AW
LaGattuta
M
M
Reddy
R G
RG
D001093:Arachidonate 15-Lipoxygenase / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008041:Linoleic Acids / Q000096:biosynthesis* / Q000502:physiology; D008297:Male; D016133:Polymerase Chain Reaction; D019048:Prostatic Intraepithelial Neoplasia; D011471:Prostatic Neoplasms / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D014407:Tumor Cells, Cultured
D008041:Linoleic Acids; C024348:13-hydroxy-9,11-octadecadienoic acid; D001093:Arachidonate 15-Lipoxygenase