The effects of PIB5PA on migration and invasion of human melanoma cells

Objective: To investigate the effects of phosphatidylinositol 4, 5-bisphosphate 5-phosphatase A (PIB5PA) on the migration and invasion of human melanoma Mel-FH cells. Methods: Eukaryotic expression recombinant vector pF-5xUAS-SV40-PIB5PA which carried a 4-hydroxytamoxifen (4-OHT)-inducible lentiviral expression system was constructed and infected into melanoma Mel-FH cells. Mel-FH.PIB5PA cells were successfully obtained via dual antibiotic selection of puromycin and hygromycin B. The optimum concentration and acting time of 4-OHT on the expression of PIB5PA of Mel-FH.PIB5PA cells were detected by Western blotting. The migration and invasion of Mel-FH.PIB5PA cells were determined by wound healing and Transwell chamber assay, respectively. The expression levels of phospho-protein kinase B (p-AKT), phospho-focal adhesion kinase (p-FAK), matrix metalloproteinase-2 (MMP-2), MMP-9, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and TIMP-2 were measured by Western blotting. Results: The addition of 10 nmol/L 4-OHT for 16 h readily induced PIB5PA overexpression in Mel-FH.PIB5PA cells. The exogenous expression of PIB5PA significantly inhibited migration and invasion of Mel-FH.PIB5PA cells as compared with Mel-FH.PIB5PA cells without treatment of 4-OHT (P < 0.05), and reduced the expression levels of p-AKT, p-FAK, MMP-2, MMP-9, TIMP-1, TIMP-2, MMP-2/ TIMP-2 and MMP-9/TIMP-1 proteins (all P < 0.05). Conclusion: Over-expression of PIB5PA can inhibit the abilities of migration and invasion of human melanoma Mel-FH cells in vitro , which may be associated with inactivity of AKT and FAK, and down-regulation of the relative expression levels of MMP-2/TIMP-2 and MMP-9/TIMP-1.