Journal: Scientific Reports

Article Title: DGAT1 Inhibitor Suppresses Prostate Tumor Growth and Migration by Regulating Intracellular Lipids and Non-Centrosomal MTOC Protein GM130

doi: 10.1038/s41598-019-39537-z

Figure Lengend Snippet: DGAT1 inhibition decreases ncMTOC-related proteins in prostate cancer cells. ( A ) LNCaP and PC-3 cells were treated with 1 µM DGAT1 inhibitor for 24 h, and, after fixation, the cells were stained with mouse anti-pericentrin antibody (green) and DAPI (blue) to visualize the MTOCs and nucleus, respectively. Size bar: 10 µm. ( B ) The total number of pericentrin-positive MTOCs was counted per single cell (n = 50). ( C ) γ-tubulin (48 kD), ninein (243 kD), GM130 (140 kD), and CETN1 (20 kD) levels were analyzed by western blot in PC-3 control (CTR) and treated with 1 µM DGAT1 inhibitor or 10 nM PEDF. ( D ) γ-tubulin, ( E ) ninein, ( F ) GM130, and ( G ) centrin 1 (CETN1) densities were normalized by GAPDH. For the western blot analysis, each protein has been detected by sequential staining and stripping using the same membrane. Data are presented as mean ± SEM. Student’s unpaired t test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: PEDF (1:1000 - Cat. No. AB-PEDF4 - BioProducts MD, Middletown, MD), ATGL (1:200 – Cat. No. 10006409 - Cayman Chemical, Ann Arbor, MI), DGAT1 (1:10000 – Cat. No. ab181180 - Abcam, Cambridge, United Kingdom), perilipin 2 (PLIN2) (2 μg/ml – Cat. No. LS-B3121 – Lifespan Biosciences, Seattle, WA), γ-tubulin (1 μg/ml – Cat. No. ab27074 - Abcam, Cambridge, United Kingdom), centrin 1 (CETN1) (1:2000 – Cat. No. ab11257 - Abcam, Cambridge, United Kingdom), ninein (1:5000 – Cat. No. NB100-74631 – Novus Biologicals, Littleton, CO), GM130 (1:5000 – Cat. No. ab52649 - Abcam, Cambridge, United Kingdom), CLASP2 (1:1000 – Cat. No. ab95373 - Abcam, Cambridge, United Kingdom), and α-tubulin (1 μg/ml – Cat. No. ab15246 - Abcam, Cambridge, United Kingdom) antibodies were used.

Techniques: Inhibition, Staining, Western Blot, Control, Stripping Membranes, Membrane

Journal: Scientific Reports

Article Title: DGAT1 Inhibitor Suppresses Prostate Tumor Growth and Migration by Regulating Intracellular Lipids and Non-Centrosomal MTOC Protein GM130

doi: 10.1038/s41598-019-39537-z

Figure Lengend Snippet: Model of DGAT1 inhibition of lipid-MTOC axis in prostate cancer. This is a model of lipid-MTOC crosstalk in prostate cancer. At baseline, PCa cells are lipid-rich with MTOC amplification and a complex and dense MT network. At the site of the Golgi apparatus, GM130 and CLASP2-coated MTs are strongly expressed. PC-3 cells express high levels of DGAT1 and low levels of PEDF protein. After treatment with a DGAT1 inhibitor, the LD density is reduced as well as the number of MTOCs. The expression levels of proteins regulating ncMTOCs (GM130 and CLASP2) and MTOCs in general (γ-tubulin and ninein) were significantly decreased. Structural changes in the MT network post-treatment revealed fragmentation and loss of many linear MTs. Blockade of the lipid-MTOC axis via DGAT1 inhibitor resulted in suppression of prostate cancer growth and invasion both in vitro and in vivo .

Article Snippet: PEDF (1:1000 - Cat. No. AB-PEDF4 - BioProducts MD, Middletown, MD), ATGL (1:200 – Cat. No. 10006409 - Cayman Chemical, Ann Arbor, MI), DGAT1 (1:10000 – Cat. No. ab181180 - Abcam, Cambridge, United Kingdom), perilipin 2 (PLIN2) (2 μg/ml – Cat. No. LS-B3121 – Lifespan Biosciences, Seattle, WA), γ-tubulin (1 μg/ml – Cat. No. ab27074 - Abcam, Cambridge, United Kingdom), centrin 1 (CETN1) (1:2000 – Cat. No. ab11257 - Abcam, Cambridge, United Kingdom), ninein (1:5000 – Cat. No. NB100-74631 – Novus Biologicals, Littleton, CO), GM130 (1:5000 – Cat. No. ab52649 - Abcam, Cambridge, United Kingdom), CLASP2 (1:1000 – Cat. No. ab95373 - Abcam, Cambridge, United Kingdom), and α-tubulin (1 μg/ml – Cat. No. ab15246 - Abcam, Cambridge, United Kingdom) antibodies were used.

Techniques: Inhibition, Amplification, Expressing, In Vitro, In Vivo