4-O-Methylascochlorin-Mediated BNIP-3 Expression Controls the Balance of Apoptosis and Autophagy in Cervical Carcinoma Cells
Abstract
:1. Introduction
2. Results
2.1. MAC Is Cytotoxic and Inhibits the Proliferation of Cervical Carcinoma Cells
2.2. MAC Induces Apoptosis of Cervical Carcinoma Cells
2.3. MAC Facilitates Autophagosome-Lysosome Fusion and Induces Autophagy in Cervical Carcinoma Cells
2.4. BNIP-3 Mediates MAC-Induced Autophagy and Apoptosis in Cervical Carcinoma Cells
2.5. Inhibition of Apoptosis Enhances MAC-Induced Autophagy in Cervical Carcinoma Cells
2.6. Inhibition of Autophagosome-Lysosome Fusion Enhances MAC-Induced Apoptosis in Cervical Carcinoma Cells
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Materials
4.2. Cell Viability Assay
4.3. Colony Formation Assay
4.4. Cell Cycle Analysis
4.5. Trypan Blue Staining
4.6. Hoechst 33,342 and PI Double Staining
4.7. Flow Cytometry for Apoptosis
4.8. RT-PCR
4.9. Western Blot Analysis
4.10. Immunofluorescence
4.11. GFP-RFP-LC3 Plasmid Transfection and Autophagy Flux
4.12. siRNA Transfection
4.13. Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
MAC | 4-O-methylascochlorin |
Akt | Serine-threonine kinase B |
mTOR | mammalian target of rapamycin |
p70S6K | 70-kDa ribosomal protein S6 Kinase |
BNIP-3 | Bcl-2/adenovirus E1B 19 kDa protein-interacting protein 3 |
HIF-1α | Hypoxia-inducible factor-1 alpha |
PARP | Poly (ADP-ribose) polymerase |
CQ | Chloroquine |
ATG | Autophagic-related gene |
AMPK | AMP-activated protein kinase |
IC50 | Half-maximal-inhibitory concentrations |
LAMP1 | Lysosomal-associated membrane protein 1 |
TFEB | Transcription factor EB |
PI | Propidium iodide |
DMEM | Dulbecco’s Modified Eagle Medium |
RPMI | Roswell Park Memorial Institute |
FBS | Fetal bovine serum |
PBS | Phosphate-buffered saline |
FITC | Fluorescein isothiocyanate |
Trypsin EDTA | Trypsin disodium ethylenediaminetetraacetic acid |
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Cho, Y.; Jeong, Y.-J.; Song, K.-H.; Chung, I.-K.; Magae, J.; Kwon, T.K.; Choi, Y.-H.; Kwak, J.-Y.; Chang, Y.-C. 4-O-Methylascochlorin-Mediated BNIP-3 Expression Controls the Balance of Apoptosis and Autophagy in Cervical Carcinoma Cells. Int. J. Mol. Sci. 2022, 23, 15138. https://doi.org/10.3390/ijms232315138
Cho Y, Jeong Y-J, Song K-H, Chung I-K, Magae J, Kwon TK, Choi Y-H, Kwak J-Y, Chang Y-C. 4-O-Methylascochlorin-Mediated BNIP-3 Expression Controls the Balance of Apoptosis and Autophagy in Cervical Carcinoma Cells. International Journal of Molecular Sciences. 2022; 23(23):15138. https://doi.org/10.3390/ijms232315138
Chicago/Turabian StyleCho, Yuna, Yun-Jeong Jeong, Kwon-Ho Song, Il-Kyung Chung, Junji Magae, Taeg Kyu Kwon, Yung-Hyun Choi, Jong-Young Kwak, and Young-Chae Chang. 2022. "4-O-Methylascochlorin-Mediated BNIP-3 Expression Controls the Balance of Apoptosis and Autophagy in Cervical Carcinoma Cells" International Journal of Molecular Sciences 23, no. 23: 15138. https://doi.org/10.3390/ijms232315138