Cytoprotective Role of Autophagy in CDIP1 Expression-Induced Apoptosis in MCF-7 Breast Cancer Cells
Abstract
:1. Introduction
2. Results
2.1. Lysosomal Degradation of CDIP1
2.2. Induction of Autophagy by CDIP1
2.3. Protective Role of Autophagy in CDIP1 Expression-Induced Apoptosis
3. Discussion
4. Materials and Methods
4.1. Antibodies and Reagents
4.2. Plasmid Constructions
4.3. Cell Culture, DNA Transfection and Lentiviral Infection
4.4. Cell Lysate Preparation and Western Blotting
4.5. Statistical Analysis
4.6. Research Ethics
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
ADR | adriamycin |
ALG-2 | apoptosis-linked gene 2 |
BafA1 | bafilomycin A1 |
BAP31 | B-cell receptor-associated protein 31 |
CDIP1 | cell death-inducing p53-target protein 1 |
CHX | cycloheximide |
CIAP | calf intestine alkaline phosphatase |
CQ | chloroquine |
Dox | doxycycline |
Epo | epoxomicin |
ER | endoplasmic reticulum |
GAPDH | glyceraldehyde-3-phosphate dehydrogenase |
LC3B | microtubule-associated proteins 1A/1B light chain 3B |
LITAF | lipopolysaccharide-induced tumor necrosis factor-α factor homolog |
PARP1 | poly[ADP-ribose] polymerase 1 |
SIMPLE | small integral membrane protein of the late endosome |
TNF-α | tumor necrosis factor-α |
TSG101 | tumor susceptibility gene 101 |
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Mutations | Sence Primer Sequences | Antisense Primer Sequences |
---|---|---|
T27A | 5′-agccccgcccgccccaggccgtt-3′ | 5′-aacggcctggggcgggcggggct-3′ |
S31A | 5′-cccaggccgtgcctccccagc-3′ | 5′-gctggggaggcacggcctggg-3′ |
S32A | 5′-caggccgttccgccccagctgtg-3′ | 5′-cacagctggggcggaacggcctg-3′ |
S32D | 5′-caggccgttccgacccagctgtg-3′ | 5′-cacagctgggtcggaaccgcctg-3′ |
S31A/S32A | 5′-cccaggccgtgccgccccagctgtg-3′ | 5′-cacagctggggcggcacggcctggg-3′ |
S72A/T76A/Y77A | 5′-caccacacatggctgcagatggcgccgccatgcctccg-3′ | 5′-cggaggcatggcggcgccatctcagccatgtgtggtg-3′ |
H89A | 5′-ctccaggccccgccccacccatg-3′ | 5′-catgggtggggcggggcctggag-3′ |
Y94A/Y95A | 5′-cacccatgggcgccgcccccccaggg-3′ | 5′-ccctgggggggcggcgcccatgggtg-3′ |
Y100A/Y105A | 5′-ccccagggcccgccacgccagggcccgcccctggccctg-3′ | 5′-cagggccaggggcgggccctggcgtggcgggccctgggg-3′ |
H111A | 5′-cctgggggcgccacagccacag-3′ | 5′-ctgtggctgtggcgcccccagg-3′ |
T112A/T114A | 5′-ctgggggccacgcagccgcagtcctggtc-3′ | 5′-gaccaggactgcggctgcgtggcccccag-3′ |
S119A | 5′-gtcctggtccctgcaggagctgccac-3′ | 5′-gtggcagctcctgcagggaccaggac-3′ |
T123A/T124A/T126A | 5′-caggagctgccgccgcagtggcagtgctgcag-3′ | 5′-ctgcagcactgccactgcggcggcaggtcctg-3′ |
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Inukai, R.; Mori, K.; Maki, M.; Takahara, T.; Shibata, H. Cytoprotective Role of Autophagy in CDIP1 Expression-Induced Apoptosis in MCF-7 Breast Cancer Cells. Int. J. Mol. Sci. 2024, 25, 6520. https://doi.org/10.3390/ijms25126520
Inukai R, Mori K, Maki M, Takahara T, Shibata H. Cytoprotective Role of Autophagy in CDIP1 Expression-Induced Apoptosis in MCF-7 Breast Cancer Cells. International Journal of Molecular Sciences. 2024; 25(12):6520. https://doi.org/10.3390/ijms25126520
Chicago/Turabian StyleInukai, Ryuta, Kanako Mori, Masatoshi Maki, Terunao Takahara, and Hideki Shibata. 2024. "Cytoprotective Role of Autophagy in CDIP1 Expression-Induced Apoptosis in MCF-7 Breast Cancer Cells" International Journal of Molecular Sciences 25, no. 12: 6520. https://doi.org/10.3390/ijms25126520