Figure 1.
Relationships between uptake of radiolabeled acetate, FASN expression, and sensitivity to orlistat treatment in vitro.
(A) Uptake of [1-14C]acetate (upper) and levels of FASN expression (lower) in LNCaP, PC3, 22Rv1, and DU145 cells. The groups with different alphabets are significantly different (P<0.05). (B) Percent cell viability after orlistat treatment in LNCaP, PC3, 22Rv1, and DU145 cells. Cell viability is indicated as a percentage of that with 0 µM orlistat treatment. Asterisks indicate statistical significance versus treatment with 0 µM for each cell line (*P<0.05). (C) Relationship between uptake of [1-14C]acetate and FASN expression (upper), relationship between % cell viability after orlistat treatment at 12.5 µM and uptake of [1-14C]acetate (middle), and relationship between % cell viability after orlistat treatment at 12.5 µM and FASN expression (lower). Values from six independent experiments are shown. Data are expressed as means ± SD.
Figure 2.
Biodistribution of [1-14C]acetate and in vivo PET imaging with [1-11C]acetate in tumor xenograft-bearing mice.
(A) Biodistribution at 10 min and 30 min in organs (left) and each tumor (right). Data represents %ID/g, expressed as means ± SD. The groups with different alphabets are significantly different (P<0.05). (B) Small-animal PET images of [1-11C]acetate at 30 min after injection. Yellow arrows indicate tumors. S = stomach; L = liver.
Figure 3.
Effects of orlistat treatment in vivo.
(A) Growth of tumors treated with orlistat (240 mg/kg/day) or vehicle only daily for 2 weeks (left) in tumor xenograft-bearing mice. Data represents tumor volume (mm3). Representative images of treated tumors at day 14 (right). (B) Relative tumor volume versus initial tumor size normalized against each untreated tumor at the same time point. The groups with different alphabets are significantly different (P<0.05). (C) Effects of orlistat treatment on body weight.
Figure 4.
Effects of FASN inhibition by RNAi in FASN-expressing LNCaP cells.
FASN-RNAi 3128 and 3129 cells and control-RNAi cells were used. (A) Relative expression of FASN, analyzed by Western blotting analysis (left) and uptake of [1-14C]acetate (right). (B) Cell proliferation over 7 days (upper, left). Light microscopy images (upper, right). Relative migration and invasion potential in FASN-RNAi cells, as compared with control-RNAi cells (lower, left and right, respectively). Values from six independent experiments are shown. Data are expressed as means ± SD. The groups with different alphabets are significantly different (P<0.05).
Figure 5.
Time-lapse analysis showing morphological changes and movement of FASN knockdown LNCaP cells.
Data indicate images in control-RNAi cells (A) and FASN-RNAi 3128 cells (B). Images were taken every 6 h for 5 days. White arrowheads indicate an example of the formation of pseudopodia, spindle-shaped morphology, and active cell migration in control-RNAi cells. Black arrowheads indicate an example of the deficient formation of pseudopodia, round morphology, and low activity of cell migration in the FASN-RNAi 3128 cells.
Table 1.
Genes down-regulated by FASN inhibition with RNAi*.
Figure 6.
The functions of FASN in tumors, mechanism of FASN inhibition, and method for applying [1-11C]acetate PET as a predictor of outcome of FASN-targeted therapies are shown.