Journal: Theranostics

Article Title: Proteomes from AMPK-inhibited peripheral blood mononuclear cells suppress the progression of breast cancer and bone metastasis.

doi: 10.7150/thno.80294

Figure Lengend Snippet: Figure 1. Suppression of tumorigenic behaviors of breast cancer cells by CMLym-Dor. CN = control, CW = CW008 (PKA activator), Dor = Dorsomorphin (AMPK inhibitor), AICAR (AMPK activator), PBMC = peripheral blood mononuclear cells, and CM = conditioned medium. The double asterisks indicate p < 0.01. (A) Suppression of MTT-based viability of MDA-MB-231, MDA-MB-436, and MCF7 breast cancer cells by CMLym-CW and CMLym-Dor, respectively. (B&C) Suppression of scratch-based migration, and transwell invasion of MDA-MB-231 cells by CMLym-Dor. (D) Reduction in p-Src, Snail, and PDL1, together with the elevation in c-Cas3 (cleaved caspase 3) in MDA-MB-231 cells by CMLym-Dor. (E&F) Suppression of MTT-based viability, and transwell invasion of MDA-MB-231 cells by CMPBMC-Dor, which was derived from peripheral blood mononuclear cells with Dorsomorphin.

Article Snippet: Recombinant proteins for ENO1 (700 ng/mL, MBS2009113, MyBioSource, San Diego, CA, USA) and MSN (700 ng/mL, MBS2031729, MyBioSource) were given to MDA-MB-231 cells, and cells were incubated for 24 h. An AMPK inhibitor (Dorsomorphin, 50 μM, #3093, Tocris, Minneapolis, MN, USA) [38], an AMPK activator (AICAR, 50 μM, #2840, Tocris) [39], an activator of PKA signaling (CW008, 20 μM, #5495, Tocris) [40], and an activator of PI3K/Akt (YS49, 10 μM, #HY-15477, MedChem Express, NJ, USA) were applied to the cells for 24 h. Taxol (0.5 nM ~ 5 μM, #1097, Tocris) [41] was applied to breast cancer cells for 48 h. Preparation of CM.

Techniques: Control, Migration, Derivative Assay

Journal: Cancers

Article Title: Exploring the Tumor-Suppressing Potential of PSCA in Pancreatic Ductal Adenocarcinoma

doi: 10.3390/cancers15204917

Figure Lengend Snippet: Lymphocyte-derived tumor-suppressive conditioned medium (CM) and PSCA. CN = control, CM = conditioned medium, CW = CW008 as a protein kinase A (PKA) activator, and Dor = Dorsomorphin as an AMP-activated protein kinase (AMPK) inhibitor. The single and double asterisks indicate p < 0.05 and 0.01, respectively. ( A , B ) Reduction in MTT-based cell viability of PANC1 and Pa03C PDAC cells (48 h) by Jurkat T-lymphocyte-derived CM. Anti-tumor CM was generated by applying 20 μM of CW008 (PKA activator) or 50 μM of Dorsomorphin (AMPK inhibitor) for 24 h. ( C ) Dose responses of MTT-based viability of PANC1 with CW008-treated lymphocyte-derived CM (48 h). CM was condensed 10 times. ( D ) Western blot images, showing the elevated level of PSCA in CW008/Dorsomorphin-treated lymphocyte-derived CM. ( E , F ) Decrease in scratch-based motility of PANC1/Pa03C PDAC cells (24 h) by CW008/Dorsomorphin-treated lymphocyte-derived CM.

Article Snippet: Cells were treated with 20 μM CW008 (#5495, Tocris Bioscience, Minneapolis, MN, USA) as an activator of PKA signaling [ ] or 50 μM of Dorsomorphin (#3093, Tocris Bioscience) as an inhibitor of AMPK signaling [ ] for one day.

Techniques: Derivative Assay, Control, Generated, Western Blot

Journal: Cancers

Article Title: Exploring the Tumor-Suppressing Potential of PSCA in Pancreatic Ductal Adenocarcinoma

doi: 10.3390/cancers15204917

Figure Lengend Snippet: PBMC-derived tumor-suppressive CM and PSCA. The double asterisks indicate p < 0.01. CN = control, CM = conditioned medium, PBMC = peripheral blood mononuclear cell, CW = CW008 as a protein kinase A (PKA) activator, and Dor = Dorsomorphin as an AMP-activated protein kinase (AMPK) inhibitor. ( A ) Reduction in the MTT-based viability of PANC1 and Pa03C PDAC cells (48 h) by PBMC CM. The anti-tumor CM was generated by CW008 and Dorsomorphin. ( B ) Reduction in the MTT-based viability of Pa03C PDAC cells (48 h) by CW008-treated PBMC-derived CM. ( C ) Downregulation of p-Src, and elevation of cleaved caspase 3 (c-Cas) in Pa03C PDAC cells (24 h) by the treatment with CW008-treated PBMC-derived CM. ( D ) SSC-A and FSC-A plots in flow cytometric analysis of PBMCs, indicating a major population of lymphocytes together with monocytes and granulocytes. ( E ) Elevation of PSCA in CW008-treated PBMC-derived CM by ELISA. ( F ) Reduction in MTT-based viability of PANC1 and AsPC-1 PDAC cells (48 h) in response to CW008-treated PBMC-derived CM.

Article Snippet: Cells were treated with 20 μM CW008 (#5495, Tocris Bioscience, Minneapolis, MN, USA) as an activator of PKA signaling [ ] or 50 μM of Dorsomorphin (#3093, Tocris Bioscience) as an inhibitor of AMPK signaling [ ] for one day.

Techniques: Derivative Assay, Control, Generated, Enzyme-linked Immunosorbent Assay

Journal: Cancers

Article Title: Exploring the Tumor-Suppressing Potential of PSCA in Pancreatic Ductal Adenocarcinoma

doi: 10.3390/cancers15204917

Figure Lengend Snippet: Tumor selectivity and inhibitory effects on ex vivo PDAC tissues. The single asterisk indicates p < 0.05. CM = conditioned medium, PBMC = peripheral blood mononuclear cells, CW = CW008 as a protein kinase A (PKA) activator, and Dor = Dorsomorphin as an AMP-activated protein kinase (AMPK) inhibitor. ( A ) Reduction in MTT-based viability of MSCs and PC-3 cells. ( B ) Shrinkage of human PDAC tissue fragments by PBMC CM, which was generated from the autologous peripheral blood. The anti-tumor CM was generated by CW008 and Dorsomorphin. ( C ) Proposed mechanism of the anti-tumor action of PSCA, which was in part mediated by MSLN. The CW008-treated CM derived from PBMCs displayed significant tumor-suppressive properties, leading to the downregulation of p-Src, β-catenin, and K-Ras in PDAC cells. Additionally, it exhibited cytotoxic effects and increased the levels of cleaved caspase 3, an established biomarker of apoptosis. Interestingly, the PSCA protein demonstrated a dual role in this context. While it functioned as an oncoprotein for PDAC cells, it paradoxically exhibited a tumor-suppressive effect when present in the extracellular milieu, acting as an atypical tumor-suppressing protein.

Article Snippet: Cells were treated with 20 μM CW008 (#5495, Tocris Bioscience, Minneapolis, MN, USA) as an activator of PKA signaling [ ] or 50 μM of Dorsomorphin (#3093, Tocris Bioscience) as an inhibitor of AMPK signaling [ ] for one day.

Techniques: Ex Vivo, Generated, Derivative Assay, Biomarker Discovery

Journal: Molecular metabolism

Article Title: Inhibition of somatostatin enhances the long-term metabolic outcomes of sleeve gastrectomy in mice.

doi: 10.1016/j.molmet.2024.101979

Figure Lengend Snippet: Figure 4: Long-term response of obese mice to SG followed by inhibition of somatostatin signaling. A. Change in weight of SG-cSst mice (purple triangles) and SG-saline mice (black triangles), sham-cSst mice (green circles) and sham-saline mice (black squares). p < 0.01 by treatment and by treatment surgery interaction by 3-way repeated measurement ANOVA. Error bars denote SEM. n ¼ 7,7,9,9. B. Glucose levels following an oral mixed meal tolerance test. Colors as in A. p < 0.01 by treatment 3-way repeated measurement ANOVA. Error bars denote SEM. n ¼ 7,7,9,9. C. Area under the curve (AUC) for the mixed meals tolerance test. D. Fasting plasma insulin levels at the end of the experiment. E,F. Fasting (E) and post-prandial (F) plasma GLP-1 levels at the end of the experiment. G. Post-prandial total cholesterol, HDL-cholesterol, and LDL-cholesterol in the plasma at the end of the experiment. H. Quantification of hepatic steatosis by percent of the area covered by lipid droplets in a hematoxylin and eosin staining. *,**p < 0.05, p < 0.01 by Tukey post-hoc test (A,F,G,H). ##p < 0.01 by surgery in 2-way ANOVA.

Article Snippet: Cyclosomatostatin treatment Cyclosomatostatin (cSst) was purchased from Tocris (Cat. No. 3493) and dissolved to a final concentration of 0.08% EtOH and saline. cSst was injected subcutaneously at 30 mg/kg body weight every evening from 7 days after the surgery until the end of the experiment.

Techniques: Inhibition, Saline, Clinical Proteomics, Staining