Hypoxia Regulates DPP4 Expression, Proteolytic Inactivation, and Shedding from Ovarian Cancer Cells
Abstract
:1. Introduction
2. Results
2.1. DPP4 Expression is Upregulated by Hypoxia in Ovarian Cancer Cells
2.2. Chronic Hypoxia Induces DPP4 Shedding from the Surface of Ovarian Cancer Cells
2.3. Chronic Hypoxic Growth Stimulates Matrix Metalloproteinase Expression and Alters DPP4 Release from Cells
2.4. MMP10 and MMP13 Mediate DPP4 Expression and Release from Cancer Cells
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Quantitative Real-Time PCR (qRT-PCR)
4.3. Enzyme Linked Immunosorbent Assay (ELISA)
4.4. DPP4 Enzyme Activity Assays
4.5. Protease Arrays
4.6. Targeted Knockdown of Selected MMPs Using shRNA
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Gene | Accession Number | Primer Sequences 5′–3′ | |
---|---|---|---|
Forward | Reverse | ||
18S | NR_003286.4 | GTAACCCGTTGAACCCCATT | CCATCCAATCGGTAGTAGCG |
HIF-1α | NM_001530 | CAGCTATTTGCGTGTGAGGA | CCTCATGGTCACATGGATGA |
DPP4 | NM_001935 | ATGCCAGGAGGAAGGAATCT | TATAGAGGGGCAGACCAGGA |
MMP1 | NM_002421 | GGACAACTCTCCTTTTGATGGA | CAAAGCCCCGATATCAGTAGAA |
MMP10 | NM_002425 | CACAGTTTGGCTCATGCCTA | TGCCATTCACATCATCTTGC |
MMP13 | NM_002427 | GACCCTGGAGCACTCATGTT | TCCTCGGAGACTGGTAATGG |
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Moffitt, L.R.; Bilandzic, M.; Wilson, A.L.; Chen, Y.; Gorrell, M.D.; Oehler, M.K.; Plebanski, M.; Stephens, A.N. Hypoxia Regulates DPP4 Expression, Proteolytic Inactivation, and Shedding from Ovarian Cancer Cells. Int. J. Mol. Sci. 2020, 21, 8110. https://doi.org/10.3390/ijms21218110
Moffitt LR, Bilandzic M, Wilson AL, Chen Y, Gorrell MD, Oehler MK, Plebanski M, Stephens AN. Hypoxia Regulates DPP4 Expression, Proteolytic Inactivation, and Shedding from Ovarian Cancer Cells. International Journal of Molecular Sciences. 2020; 21(21):8110. https://doi.org/10.3390/ijms21218110
Chicago/Turabian StyleMoffitt, Laura R., Maree Bilandzic, Amy L. Wilson, Yiqian Chen, Mark D. Gorrell, Martin K. Oehler, Magdalena Plebanski, and Andrew N. Stephens. 2020. "Hypoxia Regulates DPP4 Expression, Proteolytic Inactivation, and Shedding from Ovarian Cancer Cells" International Journal of Molecular Sciences 21, no. 21: 8110. https://doi.org/10.3390/ijms21218110