PLD-Specific Small-Molecule Inhibitors Decrease Tumor-Associated Macrophages and Neutrophils Infiltration in Breast Tumors and Lung and Liver Metastases
Fig 1
Inhibitory effects of FIPI or VU0155072-2 “NOPT” in mouse mammary tumors following xenotransplantation of human breast cancer cells.
(A-C) SCID mice were xenotransplanted with human MDA-MB-231 breast cancer cells following implantation of Alzet pumps containing 2 different PLD-small molecule inhibitors of activity: 5-fluoro-2-indolyl des-chlorohalopemide (FIPI) or N- [2- (4- oxo- 1- phenyl- 1, 3, 8- triazaspiro[4, 5]dec- 8- yl)ethyl]- 2- naphthalenecarboxamide (VU0155072-2) [also referred to as “NOPT” [31,40–42]]. Approximately 45 days post xenotransplantation, primary and axillary breast tumors, lungs and livers were harvested from mice that received MDA-MB-231 cancer cells and fixed in formalin, which were then embedded in paraffin, sectioned and mounted onto glass microscope slides. (D,E) Negative effect of small molecule inhibitors on PLD2 in mouse primary mammary and metastatic axillary tumors. (F,G) Images were quantified in terms of total mean fluorescence per field of view for PLD2 using ImageJ software. (H) PLD enzyme activity and (I) PLD2 gene expression. (J) PLD2 protein expression and (K) densitometry of same Western blot. Results are mean relative PLD gene expression (-fold) ± SEM, or mean PLD activity ± SEM, in terms of percent of control (100% = 1,254 ± 117 dpm/mg protein). Sample sizes were n = 5 for each set of animals used and 6–8 fields were viewed for each section. Statistical analyses for the increases (*, p<0.05, ANOVA) in xenotransplants and the decreases (#, p<0.05, ANOVA) in PLD inhibitors-treated samples.