Differential effects of sPLA₂-GV and GX on cellular proliferation and lipid accumulation in HT29 colon cancer cells

Secretory phospholipase A₂ (sPLA₂) group of enzymes have been shown to hydrolyze phospholipids, among which sPLA₂ Group V (GV) and Group X (GX) exhibit high selectivity towards phosphatidylcholine-rich cellular plasma membranes. The enzymes have recently emerged as key regulators in lipid droplets formation and it is hypothesized that sPLA₂-GV and GX enhanced cell proliferation and lipid droplet accumulation in colon cancer cells (HT29). In this study, cell viability and lipid droplet accumulation were assessed by Resazurin assay and Oil-Red-O staining. Interestingly, both sPLA₂-GV and GX enzymes reduced intracellular lipid droplet accumulation and did not significantly affect cell proliferation in HT29 cells. Incubation with varespladib, a pan-inhibitor of sPLA₂-Group IIA/V/X, further suppressed lipid droplets accumulation in sPLA₂-GV but have no effects in sPLA₂-GX-treated cells. Further studies using catalytically inactive sPLA₂ enzymes showed that the enzymes intrinsic catalytic activity is required for the net reduction of lipid accumulation. Meanwhile, inhibition of intracellular phospholipases (iPLA₂-γ and cPLA₂-α) unexpectedly enhanced lipid droplet accumulation in both sPLA₂-GV and GX-treated cells. The findings suggested an interconnected relationship between extracellular and intracellular phospholipases in lipid cycling. Previous studies indicated that sPLA₂ enzymes are linked to cancer development due to their ability to induce release of arachidonic acid and eicosanoids as well as the stimulation of lipid droplet formation. This study showed that the two enzymes work in a distinct manner and they neither confer proliferative advantage nor enhanced the net lipid droplet accumulation in HT29 cells.