Inactivation of APC Induces CD34 Upregulation to Promote Epithelial-Mesenchymal Transition and Cancer Stem Cell Traits in Pancreatic Cancer
Abstract
:1. Introduction
2. Results
2.1. Genetically Engineered Mouse Models (GEMMs) Recapitulate the Loss of APC in PDAC to Evoke Metastatic PDAC Development
2.2. Inactivation of APC in PDAC Induced EMT to Increase PDAC Cell Tumorigenic and Migratory Abilities
2.3. Activation of CD34 Pathway in PKAP+ PDAC Cells
2.4. Knockdown of CD34 in PKAP+ Tumor Cells Reduces Cell Invasion and Migration
2.5. Overexpression CD34 Increases Tumorigenic and Cell Migration Abilities in PDAC
3. Discussion
4. Materials and Methods
4.1. Genetically Modified Mice and Mouse Genotyping
4.2. Immunohistochemistry (IHC) and Immunofluorescence (IF)
4.3. Western Blot Analysis
4.4. Real-Time–Quantitative PCR Analysis (RT–qPCR)
4.5. Cell Proliferation Assay
4.6. Colony Formation and Hanging Drop Assays
4.7. Wound Healing Assay
4.8. Flow Cytometry Analysis
4.9. Complementary DNA Microarray Analysis
4.10. Murine Primary PDAC Cell Culture, Cytokine and Inhibitor Treatment
4.11. Retroviral Production and Infection of Target Cells
4.12. Lentivirus Production and shRNA for Gene Knockdown
4.13. Intracardiac Injection of Metastatic Tumor Model
4.14. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
PDAC | Pancreatic ductal adenocarcinoma |
EMT | Epithelial–mesenchymal transition |
IF | Immunofluorescence |
IHC | Immunohistochemistry |
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Hsieh, M.J.; Chiu, T.-J.; Lin, Y.C.; Weng, C.-C.; Weng, Y.-T.; Hsiao, C.-C.; Cheng, K.-h. Inactivation of APC Induces CD34 Upregulation to Promote Epithelial-Mesenchymal Transition and Cancer Stem Cell Traits in Pancreatic Cancer. Int. J. Mol. Sci. 2020, 21, 4473. https://doi.org/10.3390/ijms21124473
Hsieh MJ, Chiu T-J, Lin YC, Weng C-C, Weng Y-T, Hsiao C-C, Cheng K-h. Inactivation of APC Induces CD34 Upregulation to Promote Epithelial-Mesenchymal Transition and Cancer Stem Cell Traits in Pancreatic Cancer. International Journal of Molecular Sciences. 2020; 21(12):4473. https://doi.org/10.3390/ijms21124473
Chicago/Turabian StyleHsieh, Mei Jen, Tai-Jan Chiu, Yu Chun Lin, Ching-Chieh Weng, Yu-Ting Weng, Chang-Chun Hsiao, and Kuang-hung Cheng. 2020. "Inactivation of APC Induces CD34 Upregulation to Promote Epithelial-Mesenchymal Transition and Cancer Stem Cell Traits in Pancreatic Cancer" International Journal of Molecular Sciences 21, no. 12: 4473. https://doi.org/10.3390/ijms21124473
APA StyleHsieh, M. J., Chiu, T. -J., Lin, Y. C., Weng, C. -C., Weng, Y. -T., Hsiao, C. -C., & Cheng, K. -h. (2020). Inactivation of APC Induces CD34 Upregulation to Promote Epithelial-Mesenchymal Transition and Cancer Stem Cell Traits in Pancreatic Cancer. International Journal of Molecular Sciences, 21(12), 4473. https://doi.org/10.3390/ijms21124473