Down-Regulation of Transglutaminase 2 Stimulates Redifferentiation of Dedifferentiated Chondrocytes through Enhancing Glucose Metabolism
Abstract
:1. Introduction
2. Results
2.1. TG2 Expression is Associated With Dedifferentiation of Chondrocytes
2.2. Alteration of Energy Metabolism in Pellet Culture System
2.3. Down-Regulation of TG2 Enhances Glycolytic Pathway in Chondrocytes
3. Discussion
4. Materials and Methods
4.1. Ethics Statement
4.2. Chondrocyte Isolation
4.3. Pellet Preparation and Cultivation
4.4. Western Blot Analysis
4.5. RNA Isolation and Real Time Quantitative PCR
4.6. Lactate Assay
4.7. Knockdown Experiment
4.8. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
3D culture system | Three-Dimensional Culture System |
ACI | Autologous Chondrocyte Implantation |
ECM | Extracellular Matrix |
GAG | Glycosaminoglycan |
TG2 | Transglutaminase 2 |
SOX9 | SRY (Sex Determining Region Y)-Box 9 (SOX9) |
COL2A1 | Collagen Type II |
ACAN | Aggrecan |
GLUT1 | Glucose Transporter 1 |
HK2 | Hexokinase 2 |
LDHA | Lactate Dehydrogenase A |
PKM2 | Pyruvate Kinase Muscle isozyme M2 |
TGF-β1 | Transforming Growth Factor Beta 1 |
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Gene | Human Primer Sequence | |
---|---|---|
TG2 | Sense | 5′-GGCACCAAGTACCTGCTCA-3′ |
Antisense | 5′-AGAGGATGCAAAGAGGAACG-3′ | |
ACAN | Sense | 5′-GCCTGCGCTCCAATGACT-3′ |
Antisense | 5′-ATGGAACACGATGCCTTTCAC-3′ | |
COL2A1 | Sense | 5′-CACGTACACTGCCCTGAAGGA-3′ |
Antisense | 5′-CGATAACAGTCTTGCCCCACTT-3′ | |
SOX9 | Sense | 5′-CCCCAACAGATCGCCTACAG-3′ |
Antisense | 5′-GAGTTCTGGTCGGTGTAGTC-3′ | |
GLUT1 | Sense | 5′-GGTTGTGCCATACTCATGACC-3′ |
Antisense | 5′-CAGATAGGACATCCAGGGTAGC-3′ | |
HK2 | Sense | 5′-TCCCCTGCCACCAGACTA-3′ |
Antisense | 5′-TGGACTTGAATCCCTTGGTC-3′ | |
PKM2 | Sense | 5′-CGTCTGAACTTCTCTCATGGAA-3′ |
Antisense | 5′-ATGGGGTCAGAAGCAAAGC-3′ | |
LDHA | Sense | 5′-GCAGATTTGGCAGAGAGTATAATG-3′ |
Antisense | 5′-GACATCATCCTTTATTCCGTAAAGA-3′ | |
18S rRNA | Sense | 5′-CTTCCACAGGAGGCCTACAC-3′ |
Antisense | 5′-CGCAAAATATGCTGGAACTTT-3′ |
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Ko, K.-W.; Choi, B.; Park, S.; Arai, Y.; Choi, W.C.; Lee, J.-M.; Bae, H.; Han, I.-B.; Lee, S.-H. Down-Regulation of Transglutaminase 2 Stimulates Redifferentiation of Dedifferentiated Chondrocytes through Enhancing Glucose Metabolism. Int. J. Mol. Sci. 2017, 18, 2359. https://doi.org/10.3390/ijms18112359
Ko K-W, Choi B, Park S, Arai Y, Choi WC, Lee J-M, Bae H, Han I-B, Lee S-H. Down-Regulation of Transglutaminase 2 Stimulates Redifferentiation of Dedifferentiated Chondrocytes through Enhancing Glucose Metabolism. International Journal of Molecular Sciences. 2017; 18(11):2359. https://doi.org/10.3390/ijms18112359
Chicago/Turabian StyleKo, Kyoung-Won, Bogyu Choi, Sunghyun Park, Yoshie Arai, Won Chul Choi, Joong-Myung Lee, Hojae Bae, In-Bo Han, and Soo-Hong Lee. 2017. "Down-Regulation of Transglutaminase 2 Stimulates Redifferentiation of Dedifferentiated Chondrocytes through Enhancing Glucose Metabolism" International Journal of Molecular Sciences 18, no. 11: 2359. https://doi.org/10.3390/ijms18112359