Prostate cancer is the commonest male cancer in the west, with mortality second to that of lung cancer. The role played by lipogenesis in cancer biology is receiving increasing interest in the research arena (Kuhajda, 2006) whilst in prostate cancer fatty acid synthase is considered to be a metabolic oncogen (Baron et al., 2004). Understanding lipogenesis and citrate-related cytoplasmic metabolism in prostate cancer cells formed the basis of the present research (Mycielska et al., 2006). In particular, the project studied the role played by citrate metabolism, via the c-aconitase/cICD pathway, in lipogenesis. We adopted PNT2-C2 and PC-3M cell lines to represent normal and highly metastatic prostatic epithelia, respectively. Spectrophotometric assays were used to study NADPH production, free fatty levels and cICD activity in these cell lines, under various conditions. Real-time PCR was used to study mRNA expression of the major NADPH producing enzymes. NADPH levels were significantly higher in cancer cells compared to normal prostate cells (0.025 ± 0.0066 and 0.015 ± 0.0047 mmols/100 μg protein, respectively). In the presence of oxalomalate, inhibitor of the c- (cytosolic) aconitase/cICD pathway, the NADPH levels in cancer were reduced to those seen in normal prostate cells. A similar reduction in NADPH levels was seen when cancer cells were treated with an iron chelator (DFO). C-aconitase, known also as iron regulatory protein (IRP), acquires enzymatic activity in the presence of high intracellular iron, known to be associated with cancer. Therefore, DFO was used to inhibit c-aconitase activity. The same inhibitors (2.5 mM oxalomalate and 0.5 mM DFO) reduced significantly the synthesis of free fatty acids in prostate cancer cells (37.7 ± 1.1 % and 28.5 ± 8.5 % respectively) but had no effect on normal cells. cICD mRNA expression was 38 ± 0.7 % higher in PNT2-C2 cells compared to PC-3M cells, however activity of cICD measured as rate of NADPH production was strongly dependent on the inhibition/activity of FAS. In the presence of FAS inhibitors the level of NADPH resulting from cICD activity was significantly higher, by 20 ± 0.7 %, in PC-3M cells treated with inhibitor. The obtained results would suggest that: (1) c-aconitase/cICD pathway becomes active in cancer cells (2) c-aconitase/cICD provides NADPH to FAS for de novo fatty acid synthesis (3) c-aconitase activity is dependent on the intracellular levels of Fe.