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Figure 6 | Molecular Cancer

Figure 6

From: Prosaposin down-modulation decreases metastatic prostate cancer cell adhesion, migration, and invasion

Figure 6

Effect of ceramide on PCa cell adhesion, migration, and invasion. (A) PSAP down-modulation increased Cer levels in PSAP-KD cells. PSAP-KD and control clones of PC-3 and DU-145 cells were subjected to matrix-assisted laser desorption mass spectrometric analysis as described in "Materials and Methods". The assay was performed in duplicate and repeated twice independently. Cer content was quantified and calibrated to the intracellular phosphate (Pi) level and depicted as Cer (pM)/Pi (nM). (B) Effect of Cer on β1A-integrin expression. Control clones of PC-3 and DU-145 cells were treated with active or inactive Cer analogs or vehicle (DMSO) at the indicated concentrations for 36 h. Cell lysates were subjected to immunoblotting with specific antibodies against total β1-integrin or β1A-isoform. Inactive Cer analog and DMSO did not affect integrins expression level. GAPDH antibody was used as control loading. (C and D) Effect of Cer on PCa cell adhesion. Parental PC-3 and DU-145 cells were treated with cell permeable natural Cer analog (C6-D-e-Cer; D-Cer), inactive Cer (C6-L-e-Cer; L-Cer), or vehicle (DMSO) at 1 or 2 μM for three to five days and then, subjected to cell adhesion on FN- or LN-coated plates as described in the legend to Fig. 1. (E and F) Effect of Cer on PCa cell migration and invasion. The effect of Cer on migration and invasion of control PC-3 and DU-145 transfectants was examined by treating cells with active or inactive Cer analogs (described above) followed by migration and invasion assay as explained in the legend to Fig. 2. Each bar represents the mean ± SEM of three independent experiments. ANOVA models with Dunnett and Tukey corrections were used to compare cells treated with DMSO, L-Cer, or D-Cer. Statistically significant differences were set at p < 0.05. Consistent data were obtained from three independent experiments.

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