KCP10043F Represses the Proliferation of Human Non-Small Cell Lung Cancer Cells by Caspase-Mediated Apoptosis via STAT3 Inactivation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemical and Reagents
2.2. Cell Culture
2.3. Cytotoxicity Assay
2.4. Annexin V-FITC (Fluorescein Isothiocyanate) and Propidium Iodide (PI) Double Staining Assay
2.5. DAPI (4′,6-Diamidino-2-Phenylindole) Staining Assay
2.6. Terminal Deoxynucleotidyl Transferase dUTP Nick end Labeling (TUNEL) Assay
2.7. Western Blot Analysis
2.8. Mitochondrial and Cytosolic Fractionation
2.9. Measurement of Mitochondrial Membrane Potential (ΔΨm)
2.10. Transfection for Signal Transducer and Activator of Transcription (STAT3) Overexpression
2.11. Cytokine Production
2.12. Immunocytochemistry
2.13. Animals
2.14. In Vivo Tumor Xenograft Studies
2.15. Molecular Docking Analysis
2.16. Statistical Analysis
3. Results
3.1. KCP10043F Inhibits the Proliferation of A549 and NCI-H358 Human Non-Small Cell Lung Cancer (NSCLC) Cells by Inducing Apoptosis
3.2. KCP10043F Induces Caspase-Dependent Apoptosis
3.3. KCP10043F Induces the Release of Cytochrome C into the Cytosol and Loss of ΔΨm
3.4. KCP10043F Affects the STAT3 Signaling Pathway in A549 and NCI-H358 Cells
3.5. KCP10043F Docks into the SH2 Domain of STAT3
3.6. KCP10043F Suppresses Tumor Growth through Apoptotic Cell Death in A549 Xenograft Model
4. Discussion
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Lee, J.-H.; Lee, H.-H.; Ryu, K.D.; Kim, M.; Ko, D.; Chung, K.-S.; Hassan, A.H.E.; Lee, S.H.; Lee, J.Y.; Lee, K.-T. KCP10043F Represses the Proliferation of Human Non-Small Cell Lung Cancer Cells by Caspase-Mediated Apoptosis via STAT3 Inactivation. J. Clin. Med. 2020, 9, 704. https://doi.org/10.3390/jcm9030704
Lee J-H, Lee H-H, Ryu KD, Kim M, Ko D, Chung K-S, Hassan AHE, Lee SH, Lee JY, Lee K-T. KCP10043F Represses the Proliferation of Human Non-Small Cell Lung Cancer Cells by Caspase-Mediated Apoptosis via STAT3 Inactivation. Journal of Clinical Medicine. 2020; 9(3):704. https://doi.org/10.3390/jcm9030704
Chicago/Turabian StyleLee, Jeong-Hun, Hwi-Ho Lee, Ki Deok Ryu, Misong Kim, Dohyeong Ko, Kyung-Sook Chung, Ahmed H.E. Hassan, Seung Hyeun Lee, Jae Yeol Lee, and Kyung-Tae Lee. 2020. "KCP10043F Represses the Proliferation of Human Non-Small Cell Lung Cancer Cells by Caspase-Mediated Apoptosis via STAT3 Inactivation" Journal of Clinical Medicine 9, no. 3: 704. https://doi.org/10.3390/jcm9030704