Pancreatic Cancer Cell-Conditioned, Human-Derived Primary Myotubes Display Increased Leucine Turnover, Increased Lipid Accumulation, and Reduced Glucose Uptake
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Ethical Approval
2.3. Cell Culturing
2.3.1. PANC-1 Cells
2.3.2. hPECs
2.3.3. Skeletal Muscle Cells
2.4. PANC-1- and hPEC-Conditioned Myotubes
2.5. Scintillation Proximity Assay (SPA)
2.6. Substrate Oxidation Assay
2.7. Thin-Layer Chromatography and Measurement of Acid-Soluble Metabolites (ASM)
2.8. Glycogen Synthesis
2.9. Immunoblotting
2.10. RNA Isolation and Analysis of Gene Expression by qPCR
2.11. Proteomics
2.12. Presentation of Data and Statistics
3. Results
3.1. Measurement of Leucine Incorporation in Myotubes Indicated Decreased Protein Synthesis, Increased Protein Decay, and Lower mRNA Expression of MYH2 after Being Conditioned with the Medium from Pancreatic Cancer Cells
3.2. Myotubes Conditioned with PANC-1 Culture Medium Had Lower Glucose Uptake and Oxidation Compared to hPEC-Conditioned Myotubes
3.3. Akt Phosphorylation Was Higher under Basal Conditions and When Treated with Insulin in Myotubes Conditioned with PANC-1 and hPEC Culture Media
3.4. Myotubes Exposed to PANC-1-Conditioned Medium Showed Increased Lipid Accumulation and Responded Less to Electron Chain Complex Inhibitors Compared to hPEC-Conditioned Myotubes
3.5. Secretomes from hPECs and PANC-1 Cells Indicate Increased Secretion of Certain Cytokines, Growth Factors, and Growth-Factor-Binding Proteins Potentially Affecting Metabolism in Skeletal Muscle Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Protein | log10 Fold-Change PANC-1 Cells vs. hPECs | |
---|---|---|
VGF | Neurosecretory protein VGF | 3.175 |
KITLG | Kit ligand | 2.381 |
AREG | Amphiregulin | 1.810 |
LTBP3 | Latent-transforming growth factor beta-binding protein 3 | 1.779 |
IGFBP2 | Insulin-like growth-factor-binding protein 2 | 1.694 |
LTBP4 | Latent-transforming growth factor beta-binding protein 4 | 1.594 |
FGF19 | Fibroblast growth factor 19 | 1.510 |
TIMP1 | Metalloproteinase inhibitor 1 | 1.392 |
GMFB | Glia maturation factor beta | 1.338 |
BMP1 | Bone morphogenetic protein 1 | 1.362 |
MIF | Macrophage migration inhibitory factor | 0.887 |
IGF2 | Insulin-like growth factor II | 0.885 |
TGFB2 | Transforming growth factor beta-2 | −1.110 |
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Krapf, S.A.; Lund, J.; Saqib, A.U.R.; Bakke, H.G.; Rustan, A.C.; Thoresen, G.H.; Kase, E.T. Pancreatic Cancer Cell-Conditioned, Human-Derived Primary Myotubes Display Increased Leucine Turnover, Increased Lipid Accumulation, and Reduced Glucose Uptake. Metabolites 2022, 12, 1095. https://doi.org/10.3390/metabo12111095
Krapf SA, Lund J, Saqib AUR, Bakke HG, Rustan AC, Thoresen GH, Kase ET. Pancreatic Cancer Cell-Conditioned, Human-Derived Primary Myotubes Display Increased Leucine Turnover, Increased Lipid Accumulation, and Reduced Glucose Uptake. Metabolites. 2022; 12(11):1095. https://doi.org/10.3390/metabo12111095
Chicago/Turabian StyleKrapf, Solveig A., Jenny Lund, Awais Ur Rehman Saqib, Hege G. Bakke, Arild C. Rustan, G. Hege Thoresen, and Eili T. Kase. 2022. "Pancreatic Cancer Cell-Conditioned, Human-Derived Primary Myotubes Display Increased Leucine Turnover, Increased Lipid Accumulation, and Reduced Glucose Uptake" Metabolites 12, no. 11: 1095. https://doi.org/10.3390/metabo12111095