HSD3B1 Expression Is Upregulated by Interleukin 4 in HT-29 Colon Cancer Cells via Multiple Signaling Pathways
Abstract
:1. Introduction
2. Results
2.1. IL4 Induces HSD3B1 Expression in HT-29 Cells
2.2. STAT6 Is Involved in IL4-Regulated HSD3B1 Expression in HT-29 Cells
2.3. PI3K/AKT Signaling Is Involved in IL4-Regulated HSD3B1 Expression in HT-29 Cells
2.4. MAPKs Are Involved in IL4-Regulated HSD3B1 Expression in HT-29 Cells
2.5. IL4 Induces 3β-HSD Activity and Bioactive GC Production in HT-29 Cells
3. Discussion
4. Materials and Methods
4.1. Cells and Reagents
4.2. Plasmids
4.3. Western Blotting Analysis
4.4. RNA Extraction and Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR)
4.5. Electrophoretic Mobility Shift Assay (EMSA)
4.6. Short Hairpin RNA (shRNA) Knockdown
4.7. Progesterone Measurement
4.8. Conditioned Medium (CM) Preparation
4.9. GC Bioassay
4.10. T Cell Apoptosis Assay
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
AKT | Protein kinase B |
c-AMP | Cyclic adenosine monophosphate |
CM | Conditioned medium |
EMSA | Electrophoretic mobility shift assay |
ERK | Extracellular signal-regulated kinase |
GC | Glucocorticoid |
GR | Glucocorticoid receptor |
GSK3 | Glycogen synthase kinase 3 |
GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
3β-HSD | 3β-Hydroxysteroid dehydrogenase/isomerase |
HSD3B1 | Hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 |
IL4 | Interleukin 4 |
IRS | Insulin receptor substrate |
JNK | c-Jun N-terminal kinase |
LH | Luteinizing hormone |
mTOR | Mechanistic target of rapamycin |
MAPKs | Mitogen-activated protein kinases |
MEK | MAP kinase |
PI3K | Phosphoinositide-3-kinase |
P5 | Pregnenolone |
shRNA | Short hairpin RNA |
PKA | Protein kinase A |
StAR | Steroidogenic acute regulatory protein |
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Gene | Forward/Reverse Primer |
---|---|
GAPDH | F: 5′-AATCCCATCACCATCTTCCA-3′ R: 5′-TGGACTCCACGACGTACTCA-3′ |
GSK3A | F: 5′-GCCCACTTCCCCCTCTCTT-3′ R: 5′-GTTGAGAGACGGTTGGATGGA-3′ |
GSK3B | F: 5′-CGGTGCAGCAGCCTTCA-3′ R: 5′-TGCCGTCCTTGTCTCTGCTA-3′ |
HSD3B1 | F: 5′-CGGCTAACGGGTGGAATCTG-3′ R: 5′-CCCCATAGATATACATGGGTCGTAAG-3′ |
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Chen, H.-M.; Hung, P.-Y.; Chen, C.-H.; Yu, Y.-J.; Syu, M.-S.; Hu, M.-C. HSD3B1 Expression Is Upregulated by Interleukin 4 in HT-29 Colon Cancer Cells via Multiple Signaling Pathways. Int. J. Mol. Sci. 2022, 23, 13572. https://doi.org/10.3390/ijms232113572
Chen H-M, Hung P-Y, Chen C-H, Yu Y-J, Syu M-S, Hu M-C. HSD3B1 Expression Is Upregulated by Interleukin 4 in HT-29 Colon Cancer Cells via Multiple Signaling Pathways. International Journal of Molecular Sciences. 2022; 23(21):13572. https://doi.org/10.3390/ijms232113572
Chicago/Turabian StyleChen, Hsin-Mei, Pei-Yu Hung, Chih-Hung Chen, Yu-Jhen Yu, Ming-Shan Syu, and Meng-Chun Hu. 2022. "HSD3B1 Expression Is Upregulated by Interleukin 4 in HT-29 Colon Cancer Cells via Multiple Signaling Pathways" International Journal of Molecular Sciences 23, no. 21: 13572. https://doi.org/10.3390/ijms232113572