Journal: Life sciences

Article Title: Fluid shear stress facilitates prostate cancer metastasis through Piezo1-Src-YAP axis.

doi: 10.1016/j.lfs.2022.120936

Figure Lengend Snippet: Fig. 1. Piezo1 expression is enhanced by wall shear stress (A) The prostate tumor microenvironment was immunohistochemically stained to evaluate Piezo1 expression. Scale bars represent 250 μm and 25 μm, respectively. (B) PC3 human prostate cancer cells were plated in the microfluidics channel and applied shear stress at 0.05 dyne cm−2 for 5 h. After wall shear stress (WSS), gene expression of Piezo1 was confirmed by qRT-PCR (n = 3, independent experiments, two-tailed t- test, *** p < 0.001). (C) Immunofluorescence staining analysis was performed to evaluate Piezo1 expression (red). Cell nuclei were stained with DAPI (blue). Scale bars represent 10 μm. (D) Protein expression of Piezo1 was detected using immunoblotting at 0, 1, 5, and 12 h after WSS.

Article Snippet: PC3, PC3-Luciferase (PC3-Luc2), and DU145 human prostate cancer cells were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Expressing, Shear, Staining, Gene Expression, Quantitative RT-PCR, Two Tailed Test, Immunofluorescence, Western Blot

Journal: Life sciences

Article Title: Fluid shear stress facilitates prostate cancer metastasis through Piezo1-Src-YAP axis.

doi: 10.1016/j.lfs.2022.120936

Figure Lengend Snippet: Fig. 5. Piezo1 induces YAP activation through tyrosine 357 YAP and Src phosphorylation (A) and (B) Immunoblot analysis was measured after treatment with mock, a Piezo1 agonist (Yoda1, 1 μM), and an Src inhibitor (dasatinib, 100 nM) and exposure to static or WSS for 5 h. (C) and (D) Immunoblot analysis was measured after exposure to WSS for 5 h in PC3 cells transfected with siControl or siPiezo1. (B) and (D) Graphs showing quantification of the expression level of each phosphorylated protein compared with the expression of the non-phosphorylated isoform. (n = 3, Two Way ANOVA with Holm-Sidak multiple comparisons, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

Article Snippet: PC3, PC3-Luciferase (PC3-Luc2), and DU145 human prostate cancer cells were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Activation Assay, Phospho-proteomics, Western Blot, Transfection, Expressing

Journal: Life sciences

Article Title: Fluid shear stress facilitates prostate cancer metastasis through Piezo1-Src-YAP axis.

doi: 10.1016/j.lfs.2022.120936

Figure Lengend Snippet: Fig. 6. In vivo monitoring of orthotopic PC3 pros tate tumor growth and metastasis with or without shPiezo1. (A) Representative whole-body images of bioluminescence measurements for xenografted PC3 tumor growth and metastasis. Luciferase tag ged PC3-Luc/shCon and PC3-Luc/shPiezo1 cells were surgically implanted into the dorsal prostate of male nude mice. Ventral images of anesthetized representative sham, control (PC3-Luc/shCon), and shPiezo1 (PC3-Luc/shPiezo1) were taken weekly for 6 weeks to monitor spreading to other organs. All images were pseudo color set using the same intensity (sham = 4, shCon = 6, shPiezo1 = 6). (B) Histological slides include orthotopic tumors of PC3-luc/shCon and PC3-luc/shPiezo1. Representa tive images were derived from mouse prostate stained with hematoxylin and eosin (H&E). High magnified images of the top panel were presented as below. Scale bars = 25 μm and 100 μm, respectively.

Article Snippet: PC3, PC3-Luciferase (PC3-Luc2), and DU145 human prostate cancer cells were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: In Vivo, Luciferase, Control, Derivative Assay, Staining