A Potential Anti-Glioblastoma Compound LH20 Induces Apoptosis and Arrest of Human Glioblastoma Cells via CDK4/6 Inhibition
Abstract
:1. Introduction
2. Results
2.1. LH20 Inhibits GBM Cell Proliferation
2.2. LH20 Suppresses GBM Cell Migration
2.3. LH20 Induces GBM Cell Apoptosis and Mitochondria Membrane Potential (MMP) Drop
2.4. LH20 Arrested the GBM Cell Cycle at G2/M Phase
2.5. CDK4/6 Were Predicted to Be the Targets of LH20
2.6. LH20 Inactivates Phosphorylation of Retinoblastoma (Rb), the Downstream Protein of CDK4/6
3. Discussion
4. Materials and Methods
4.1. Preparation of LH20
4.2. Cell Culture
4.3. Cell Viability
4.4. LDH Assay
4.5. Wound Healing Assay
4.6. Annexin V-FITC/PI Apoptosis Assay
4.7. Determination of MMP
4.8. Cell Cycle Analysis
4.9. Predicting Targets and Pathways of LH20
4.10. Molecular Docking
4.11. Western Blotting Analysis
4.12. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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Antibody | Source | Identifier | Dilution Rate |
---|---|---|---|
Rabbit anti-CDK1 | Proteintech | Cat# 19532-1-AP | 1:1000 |
Rabbit anti-CDK4 | Abcam | Cat# ab199728 | 1:2000 |
Mouse anti-CDK6 | Santa Cruz Biotechnology | Cat# sc-7961 | 1:200 |
Mouse anti-Rb | Santa Cruz Biotechnology | Cat# sc-102 | 1:200 |
Rabbit anti-pRb (Ser807/811) | Cell Signaling Technology | Cat# 8516 | 1:1000 |
Mouse anti-CylinA2 | Proteintech | Cat# 66391-1-lg | 1:5000 |
Rabbit anti-GAPDH | Proteintech | Cat# 10494-1-AP | 1:6000 |
Mouse anti-Tubulin | Proteintech | Cat# 66031-1-lg | 1:20,000 |
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Wang, Y.; Li, Y.; Liu, D.; Zheng, D.; Li, X.; Li, C.; Huang, C.; Wang, Y.; Wang, X.; Li, Q.; et al. A Potential Anti-Glioblastoma Compound LH20 Induces Apoptosis and Arrest of Human Glioblastoma Cells via CDK4/6 Inhibition. Molecules 2023, 28, 5047. https://doi.org/10.3390/molecules28135047
Wang Y, Li Y, Liu D, Zheng D, Li X, Li C, Huang C, Wang Y, Wang X, Li Q, et al. A Potential Anti-Glioblastoma Compound LH20 Induces Apoptosis and Arrest of Human Glioblastoma Cells via CDK4/6 Inhibition. Molecules. 2023; 28(13):5047. https://doi.org/10.3390/molecules28135047
Chicago/Turabian StyleWang, Yan, Youbin Li, Dong Liu, Danyang Zheng, Xiaogang Li, Chang Li, Caihui Huang, Yun Wang, Xuesong Wang, Qifu Li, and et al. 2023. "A Potential Anti-Glioblastoma Compound LH20 Induces Apoptosis and Arrest of Human Glioblastoma Cells via CDK4/6 Inhibition" Molecules 28, no. 13: 5047. https://doi.org/10.3390/molecules28135047