5-hydroxy-6-8-11-14-eicosatetraenoic-acid and Ovarian-Neoplasms

5-lipoxygenase (5-LOX), a member of the lipoxygenase gene family, is a key enzyme assisting in the conversion of arachidonic acid to 5-HETE and leukotrienes. Tumor-associated macrophages (TAMs) have a critical role in the progression and metastasis of many tumors, including ovarian tumors. Moreover, TAMs are often found in a high density in the hypoxic areas of tumors. However, the relevant mechanisms have not been studied explicitly until now. In this study, we found that the expression of 5-LOX strongly correlated with the density of TAMs in hypoxic areas of human ovarian tumor tissues. In cultured ovarian cancer cells, 5-LOX metabolites were increased under hypoxic conditons. Increased 5-LOX metabolites from hypoxic ovarian cancer cells promoted migration and invasion of macrophages, which was further demonstrated to be mediated by the upregulation of matrix metalloproteinase (MMP)-7 expression through the p38 pathway. Besides, we also showed that 5-LOX metabolites enhanced the release of tumor necrosis factor (TNF-α) and heparin-binding epidermal growth factor-like growth factor through upregulation of MMP-7. Furthermore, in animal models, Zileuton (a selective and specific 5-LOX inhibitor) reduced the MMP-7 expression and the number of macrophages infiltrating in the xenograft. Our findings suggest for the first time that increased metabolites of 5-LOX from hypoxic ovarian cancer cells promote TAM infiltration. These results of this study have immediate translational implications for the therapeutic exploitation of TAMs.

Topics: Animals; Arachidonate 5-Lipoxygenase; Cell Line; Cell Line, Tumor; Chemotaxis, Leukocyte; Disease Models, Animal; Female; Heparin-binding EGF-like Growth Factor; Heterografts; Humans; Hydroxyeicosatetraenoic Acids; Hydroxyurea; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit; Leukotriene B4; Macrophages; MAP Kinase Signaling System; Matrix Metalloproteinase 7; Mice; Neoplasm Metastasis; Ovarian Neoplasms; Tumor Necrosis Factor-alpha

To understand the mechanism for the role of arachidonic acid (AA) in steroidogenic acute regulatory (StAR) gene transcription, sections of the -1/-966 StAR promoter were deleted to produce constructs of -1/-426, -1/-211, -1/-151, and -1/-110 and inserted into the PGL3 vector to drive luciferase expression. Results indicated that -1/-151 StAR promoter contains the elements that are most responsive to AA. Electrophoretic mobility shift assays using nuclear extracts from AA-treated MA-10 Leydig tumor cells showed that AA enhanced specific binding of the nuclear extract to a 30bp (-67/-96) sequence of the StAR promoter. Also, HPLC was used to identify AA metabolites involved in StAR gene transcription. It was found that 1mM N6,2-O-dibutyryladenosine 3:5-cyclic monophosphate (dbcAMP) significantly increased the 5-lipoxygenase metabolites, 5-hydroperoxyeicosatetraenoic acid (5-HPETE) and 5-hydroxyeicosatetraenoic acid (5-HETE). Moreover, in the presence of 0.2mM dbcAMP addition of 20 microM 5-HPETE or 5-HETE significantly enhanced StAR protein expression and progesterone production (P<0.05). Similar results were obtained for StAR gene transcription with StAR mRNA levels and StAR promoter activities being significantly increased (P<0.05) when 5-HPETE was added to MA-10 cell cultures. In summary, the present studies demonstrated that cyclic AMP (cAMP) stimulated the production of the AA metabolites, 5-HPETE and 5-HETE, and showed that these metabolites enhanced StAR gene expression and steroid hormone production. The results further suggested that the AA-responsive element resides in the -67/-96 region of the StAR promoter.

Topics: Animals; Arachidonate 5-Lipoxygenase; Arachidonic Acids; Base Sequence; Bucladesine; Cyclic AMP; DNA Primers; Female; Gene Expression Regulation; Genes, Reporter; Hydroxyeicosatetraenoic Acids; Kinetics; Leydig Cell Tumor; Luciferases; Membrane Proteins; Mice; Mutagenesis, Insertional; Ovarian Neoplasms; Phosphoproteins; Promoter Regions, Genetic; Recombinant Proteins; Sequence Deletion; Tumor Cells, Cultured