P4-05-04: Arachidonic Acid-Induced Elevated Expression of 5-Lipoxygenase Is Linked to Metastatic Migration of Breast Cancer Cells.

Abstract

Abstract Background: In invasive breast cancers, cancerous cells spread outside the ducts of the breast and metastasize to lung and other tissues. Although the rapid arachidonic acid (AA) metabolism and concomitant increase of eicosanoid molecules are involved in the proliferation and invasion of breast cancer cells, the exact mechanism by which AA metabolites regulate these phenomena are not well understood. Here, we show that leukotriene B4 (LTB4), one of the metabolites of AA, which is produced by the action of 5-lipoxygenase (5-LOX), causes the proliferation and metastatic migration of breast cancer cells. Material and Methods: MDA-MB-231 (invasive breast cancer cells) and MCF7 (non-invasive breast cancer cells) were used in this study. Briefly, cells were treated with AA (100 μM) and nordihydroguaiarectin acid (NDGA, 10 μM), and the secreted eicosanoids were characterized by HPLC. Immunofluorescence microscopy was performed to elucidate the expression and intracellular localization of 5-LOX. The metastatic migration was analyzed by wound-healing assays. Results: Our results suggest that while MDA-MB-231 cells produce high levels of PGE2 and PGD2, MCF7 cells synthesize excess HETE compounds (HETE5 and HETE8). Interestingly, MDA-MB-231 cells, when stimulated with AA, show the increased syntheses of LTB4 (∼3 fold) and decreased PGE2 and PGD2 (∼2 fold). In MCF7, on the contrary, AA treatment reduced the syntheses of all eicosanoids. Furthermore, the expression of 5-LOX in MDA-MB-231 cells was also increased by ∼2 fold. We observed that AA promotes cell migration in MDA-MB-231 cells, which could be blocked by NDGA, a generic inhibitor of LOX enzymes. Because LTB4 production and its binding with BLT receptors are linked to IL-8 secretion, we measured the level of IL-8 synthesized by both MCF7 and MDA-MB-231 cells. The results suggest that MDA-MB-231 cells secrete excess IL-8 (∼2 fold) when stimulated with AA, suggesting that LTB4 and IL-8 interactive pathways are important for cancer metastasis. Discussion: Our results demonstrate that LTB4 synthesis is linked to the metastatic migration of MDA-MB-231 cells, and that it therefore should be considered as a target for developing new drugs for the treatment of invasive breast cancers. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-05-04.