Altered cellular metabolism (including lipogenesis) is a hallmark of cancer development. Our group has previously found that expression of Tumor Protein D52 (TPD52), which is amplified/ overexpressed in cancers of diverse cellular origins, increases neutral lipid storage within cultured cells (1). TPD52 co-localised with Golgi but not ER markers, and also showed partial co-localisation with Adrp-coated lipid droplets (LDs). As Brefeldin A (BFA) treatment alters both Golgi structure and lipid storage, we examined BFA effects in TPD52-expressing 3T3 cells, and in human AU-565 and HMC-1-8 breast cancer cells, all of which contain prominent LDs. Five-hour BFA treatment reduced median LD numbers in TPD52-expressing 3T3 cells, but increased LD areas. TPD52 knock-down also decreased both LD areas and numbers, and blunted BFA’s effects on LD numbers per cell. In TPD52-expressing 3T3 cells treated with BFA for 1-3 hours, TPD52 co-localised with the trans-Golgi network protein syntaxin 6. However, after 4-5 hours of BFA treatment, TPD52 showed significantly increased co-localisation with LDs in each cell line. Four hours of BFA treatment also increased Tip47 but reduced Adrp detection at LDs in TPD52-expressing 3T3 cells, with Tip47 recruitment to LDs preceding that of TPD52. LD co-localisation of TPD52 but not Tip47 was disrupted by nocodazole when co-treated with BFA. TPD52 may therefore participate in a temporal hierarchy of proteins that respond to alterations in lipid storage, with TPD52 recruitment to LDs being delayed relative to other LD regulators.
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