Curcumin Promotes KLF5 Proteasome Degradation through Downregulating YAP/TAZ in Bladder Cancer Cells
Abstract
:1. Introduction
2. Results
2.1. Curcumin Down-Regulated KLF5 Protein Expression in a Dose- and Time-Dependent Manner in 5637 and WH Bladder Cancer Cells
2.2. Curcumin Promoted Proteasome-Dependent Degradation of KLF5 Protein
2.3. KLF5 Mediated the Anti-Proliferative Effect of Curcumin
2.4. Curcumin Down-Regulated YAP and TAZ Expression
2.5. YAP Played Critical Roles in KLF5 Protein Stability in 5637 Cells
2.6. Curcumin Inhibited Subcutaneous Tumor Growth and the YAP/TAZ/KLF5/Cyclin D1 Axis in Vivo
3. Discussion
4. Experimental Section
4.1. Cell Culture and Reagents
4.2. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Assay
4.3. Western Blot Analysis
4.4. Cycloheximide (CHX) Chase Assay
4.5. Plasmids, Lentivirus Preparation, siRNA and Transfection
4.6. Real-Time Quantitative PCR (qPCR)
Gene | Forward Primer | Reverse Primer |
---|---|---|
GAPDH | ATGGGGAAGGTGAAGGTCGG | GACGGTGCCATGGAATTTGC |
KLF5 | CAGAGGACCTGGTCCAGACAAG | GAGGCCAGTTCTCAGGTGAGTG |
WWP1 | ATGTTGTGTGGCATGCAGGA | CTGCAATAGTCGCATTCTTACTTCA |
FBXW7 | CATATGTTGCTCAAAGGTGGCAAG | TGGGACAACAATCCCATATTTGAAG |
SMURF2 | CAAGATTGCTTCAGTTTGTGACAG | GGGCTTTCGGCAGGTTGTT |
YAP1 | AATTTGCCCAGTTATACCTCAGTG | CACATCAAGGCTATGATTCAAACTC |
TAZ | GCTGAGCGTACAGGCAGACA | AGGAACATAAACCATGGGTCCTT |
CDK6 | TGCACAGTGTCACGAACAGA | ACCTCGGAGAAGCTGAAACA |
AXL | CGTAACCTCCACCTGGTCTC | TCCCATCGTCTGACAGCA |
ITGB2 | AAGTGACGCTTTACCTGCGAC | AAGCATGGAGTAGGAGAGGTC |
CYR61 | GGTCAAAGTTACCGGGCAGT | GGAGGCATCGAATCCCAGC |
4.7. Tumor Xenograft Model
4.8. Immunohistochemistry
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
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Gao, Y.; Shi, Q.; Xu, S.; Du, C.; Liang, L.; Wu, K.; Wang, K.; Wang, X.; Chang, L.S.; He, D.; et al. Curcumin Promotes KLF5 Proteasome Degradation through Downregulating YAP/TAZ in Bladder Cancer Cells. Int. J. Mol. Sci. 2014, 15, 15173-15187. https://doi.org/10.3390/ijms150915173
Gao Y, Shi Q, Xu S, Du C, Liang L, Wu K, Wang K, Wang X, Chang LS, He D, et al. Curcumin Promotes KLF5 Proteasome Degradation through Downregulating YAP/TAZ in Bladder Cancer Cells. International Journal of Molecular Sciences. 2014; 15(9):15173-15187. https://doi.org/10.3390/ijms150915173
Chicago/Turabian StyleGao, Yang, Qi Shi, Shan Xu, Chong Du, Liang Liang, Kaijie Wu, Ke Wang, Xinyang Wang, Luke S. Chang, Dalin He, and et al. 2014. "Curcumin Promotes KLF5 Proteasome Degradation through Downregulating YAP/TAZ in Bladder Cancer Cells" International Journal of Molecular Sciences 15, no. 9: 15173-15187. https://doi.org/10.3390/ijms150915173