Opposite Regulation of CHOP and GRP78 and Synergistic Apoptosis Induction by Selenium Yeast and Fish Oil via AMPK Activation in Lung Adenocarcinoma Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Reagents and Chemicals
2.3. Measurement of Cell Viability
2.4. Photograph of the Cells
2.5. Analysis of Apoptotic and Necrotic Cell Death by Annexin V/7-Amino-Actinomycin D Staining
2.6. Analysis of Apoptotic Sub-G1 Fraction by Propidium Iodide Staining
2.7. Western Blot
2.8. Antibodies
2.9. Analysis of Synergistic Combination Effect
2.10. Statistical Analysis
3. Results
3.1. SY and FO Act Synergistically to Inhibit the Growth of LADC Cells
3.2. SY and FO Act Synergistically to Induce Apoptosis of LADC Cells
3.3. Synergistic Activation of AMPK by SY and FO in A549 LADC Cells
3.4. Combination of SY and FO Elevates CHOP and Reduces GRP78 in A549 LADC Cells
3.5. AMPK Inhibition Diminishes the Synergistic Apoptosis Induction and Caspase-4 Activation
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AMPK | AMP-activated protein kinase |
CC | compound C |
CHOP | CCAAT/enhancer-binding protein homologous protein |
DR5 | death receptor-5 |
EGFR | epidermal growth factor receptor |
ER | endoplasmic reticulum |
FO | fish oil |
GRP78 | glucose-regulated protein 78 |
p-IRE1 | phospho-inositol-requiring enzyme 1 |
LADC | Lung adenocarcinoma |
SEM | standard error of the mean |
SY | selenium yeast |
p-JNK | phospho-Jun amino-terminal kinase |
p-PERK | phospho-protein kinase-like endoplasmic reticulum kinase |
UPR | unfolded protein response |
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SY (ng/mL) | FO (μM) | FA | CI |
---|---|---|---|
250 | 130 | 0.29 | 0.945 |
250 | 200 | 0.36 | 0.968 |
250 | 260 | 0.44 | 0.983 |
500 | 130 | 0.43 | 0.840 |
500 | 200 | 0.65 | 0.576 |
500 | 260 | 0.70 | 0.631 |
1000 | 130 | 0.51 | 0.969 |
1000 | 200 | 0.69 | 0.650 |
1000 | 260 | 0.72 | 0.711 |
2000 | 130 | 0.63 | 0.999 |
2000 | 200 | 0.72 | 0.813 |
2000 | 260 | 0.80 | 0.683 |
SY (ng/mL) | FO (μM) | FA | CI |
---|---|---|---|
125 | 130 | 0.43 | 0.740 |
125 | 200 | 0.46 | 0.565 |
125 | 260 | 0.55 | 0.738 |
250 | 130 | 0.52 | 0.974 |
250 | 200 | 0.57 | 0.950 |
250 | 260 | 0.68 | 0.775 |
500 | 130 | 0.73 | 0.990 |
500 | 200 | 0.76 | 0.950 |
500 | 260 | 0.86 | 0.664 |
1000 | 130 | 0.88 | 1.009 |
1000 | 200 | 0.88 | 1.040 |
1000 | 260 | 0.92 | 0.810 |
SY (ng/mL) | FO (μM) | FA | CI |
---|---|---|---|
250 | 130 | 0.38 | 0.856 |
250 | 200 | 0.51 | 0.347 |
250 | 260 | 0.71 | 0.468 |
500 | 130 | 0.62 | 0.607 |
500 | 200 | 0.78 | 0.389 |
500 | 260 | 0.41 | 0.993 |
1000 | 130 | 0.52 | 0.955 |
1000 | 200 | 0.72 | 0.529 |
1000 | 260 | 0.82 | 0.381 |
2000 | 130 | 0.58 | 1.208 |
2000 | 200 | 0.74 | 0.697 |
2000 | 260 | 0.85 | 0.422 |
SY (ng/mL) | FO (μM) | FA | CI |
---|---|---|---|
250 | 130 | 0.37 | 0.706 |
250 | 200 | 0.52 | 0.675 |
250 | 260 | 0.63 | 0.700 |
500 | 130 | 0.46 | 0.677 |
500 | 200 | 0.61 | 0.612 |
500 | 260 | 0.76 | 0.549 |
1000 | 130 | 0.54 | 0.694 |
1000 | 200 | 0.67 | 0.603 |
1000 | 260 | 0.8 | 0.521 |
2000 | 130 | 0.68 | 0.575 |
2000 | 200 | 0.73 | 0.604 |
2000 | 260 | 0.82 | 0.535 |
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Kao, R.-H.; Lai, G.-M.; Chow, J.-M.; Liao, C.-H.; Zheng, Y.-M.; Tsai, W.-L.; Hsia, S.; Lai, I.-C.; Lee, H.-L.; Chuang, S.-E.; et al. Opposite Regulation of CHOP and GRP78 and Synergistic Apoptosis Induction by Selenium Yeast and Fish Oil via AMPK Activation in Lung Adenocarcinoma Cells. Nutrients 2018, 10, 1458. https://doi.org/10.3390/nu10101458
Kao R-H, Lai G-M, Chow J-M, Liao C-H, Zheng Y-M, Tsai W-L, Hsia S, Lai I-C, Lee H-L, Chuang S-E, et al. Opposite Regulation of CHOP and GRP78 and Synergistic Apoptosis Induction by Selenium Yeast and Fish Oil via AMPK Activation in Lung Adenocarcinoma Cells. Nutrients. 2018; 10(10):1458. https://doi.org/10.3390/nu10101458
Chicago/Turabian StyleKao, Ruey-Ho, Gi-Ming Lai, Jyh-Ming Chow, Chien-Huang Liao, Yu-Mei Zheng, Wei-Lun Tsai, Simon Hsia, I-Chun Lai, Hsin-Lun Lee, Shuang-En Chuang, and et al. 2018. "Opposite Regulation of CHOP and GRP78 and Synergistic Apoptosis Induction by Selenium Yeast and Fish Oil via AMPK Activation in Lung Adenocarcinoma Cells" Nutrients 10, no. 10: 1458. https://doi.org/10.3390/nu10101458