Zhankuic Acids A, B and C from Taiwanofungus camphoratus Act as Cytotoxicity Enhancers by Regulating P-Glycoprotein in Multi-Drug Resistant Cancer Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals and Reagents
2.2. Cell Lines
2.3. SRB Cytotoxicity Assay
2.4. Intracellular Calcein Accumulation Assay
2.5. Real-Time Quantitative RT-PCR
2.6. MDR1 Shift Assay
2.7. Rhodamine123 and Doxorubicin Efflux Assay
2.8. P-gp ATPase Activity Assay
2.9. Cell Cycle Analysis
2.10. Apoptosis Assay
2.11. Statistical Analysis
3. Results
3.1. P-gp Efflux Function was Inhibited by ZA-A, ZA-B and ZA-C
3.2. ZA-A, ZA-B and ZA-C were not P-gp Substrates and the ABCB1 mRNA Levels were not Influenced by These Triterpenoids
3.3. The Inhibitory Mechanisms and ATPase Interactions between P-gp and ZA-A, ZA-B and ZA-C
3.4. The MDR Reversal Effects and Molecular Mechanisms of ZA-A, ZA-B and ZA-C
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Cell Line | Flp-InTM293 | ABCB1/Flp-InTM293 | ||
Compound | IC50 ± S.E. (nM) | RF | IC50 ± S.E. (nM) | RF |
Doxorubicin | 84.783 ± 0.856 | 1.0 | 767.708 ± 135.973 | 1.0 |
+ ZA-A (10 μM) | 53.276 ± 1.913 | 1.6 | 462.964 ± 27.210 | 1.7 |
+ ZA-A (20 μM) | 44.824 ± 0.436 | 1.9 | 433.570 ± 9.658 | 1.8 |
+ ZA-A (40 μM) | 37.224 ± 2.369 * | 2.3 | 124.221 ± 14.013 * | 6.2 |
+ ZA-B (10 μM) | 50.806 ± 7.431 | 1.7 | 462.296 ± 39.402 | 1.7 |
+ ZA-B (20 μM) | 48.319 ± 6.729 | 1.8 | 414.329 ± 18.101 | 1.9 |
+ ZA-B (40 μM) | 42.509 ± 2.711 * | 2.0 | 208.154 ± 3.263 * | 3.7 |
+ ZA-C (10 μM) | 64.002 ± 1.282 | 1.3 | 555.031 ± 6.192 | 1.4 |
+ ZA-C (20 μM) | 56.275 ± 2.034 | 1.5 | 507.168 ± 26.490 | 1.5 |
+ ZA-C (40 μM) | 45.570 ± 0.215 | 1.9 | 215.486 ± 105.299 * | 3.6 |
Cell Line | Flp-InTM293 | ABCB1/Flp-InTM293 | ||
Compound | IC50 ± S.E. (nM) | RF | IC50 ± S.E. (nM) | RF |
Paclitaxel | 6.556 ± 0.037 | 1.0 | 381.099 ± 19.440 | 1.0 |
+ ZA-A (10 μM) | 6.342 ± 0.336 | 1.0 | 119.886 ± 1.945 * | 3.2 |
+ ZA-A (20 μM) | 4.649 ± 0.314 | 1.4 | 33.885 ± 7.803 * | 11.2 |
+ ZA-A (40 μM) | 1.607 ± 0.638 * | 4.1 | 4.099 ± 1.729 * | 93.0 |
+ ZA-B (10 μM) | 5.749 ± 0.021 | 1.1 | 243.946 ± 26.032 | 1.6 |
+ ZA-B (20 μM) | 4.586 ± 0.182 | 1.4 | 172.364 ± 9.353 * | 2.2 |
+ ZA-B (40 μM) | 4.362 ± 0.650 | 1.5 | 27.096 ± 2.799 * | 14.1 |
+ ZA-C (10 μM) | 5.089 ± 0.752 | 1.3 | 312.457 ± 3.101 | 1.2 |
+ ZA-C (20 μM) | 4.534 ± 0.063 | 1.4 | 285.500 ± 3.030 | 1.3 |
+ ZA-C (40 μM) | 3.844 ± 0.297 | 1.7 | 201.582 ± 8.884 | 1.9 |
Cell Line | Flp-InTM293 | ABCB1/Flp-InTM293 | ||
Compound | IC50 ± S.E. (nM) | RF | IC50 ± S.E. (nM) | RF |
Vincristine | 83.822 ± 1.350 | 1.0 | 3858.781 ±217.537 | 1.0 |
+ ZA-A (10 μM) | 85.570 ± 0.120 | 1.0 | 1868.840 ± 12.138 * | 2.1 |
+ ZA-A (20 μM) | 76.274 ± 9.852 | 1.1 | 1200.585 ± 112.243 * | 3.2 |
+ ZA-A (40 μM) | 3.732 ± 0.040 * | 22.5 | 134.419 ± 4.400 * | 28.7 |
+ ZA-B (10 μM) | 43.234 ± 1.928 | 1.9 | 1698.434 ± 325.352 * | 2.3 |
+ ZA-B (20 μM) | 36.645 ± 12.895 * | 2.3 | 1475.014 ± 111.837 * | 2.6 |
+ ZA-B (40 μM) | 4.459 ± 0.455 * | 18.8 | 399.390 ± 30.490 * | 9.7 |
+ ZA-C (10 μM) | 69.245 ± 0.871 | 1.2 | 2995.200 ± 51.279 | 1.3 |
+ ZA-C (20 μM) | 54.267 ± 1.594 | 1.5 | 2274.225 ± 75.201 | 1.7 |
+ ZA-C (40 μM) | 7.801 ± 1.350 * | 10.7 | 1553.927 ± 33.018 * | 2.5 |
Cell Line | HeLaS3 | KB/VIN | ||
Compound | IC50 ± S.E. (nM) | RF | IC50 ± S.E. (nM) | RF |
Doxorubicin | 101.088 ± 1.283 | 1.0 | 3819.266 ± 98.110 | 1.0 |
+ ZA-A (10 μM) | 75.675 ± 0.732 | 1.3 | 427.791 ± 49.019 * | 8.9 |
+ ZA-A (20 μM) | 51.914 ± 1.216 | 1.9 | 76.419 ± 0.151 * | 50.0 |
+ ZA-B (10 μM) | 105.041 ± 6.633 | 1.0 | 2109.439 ± 24.634 | 1.8 |
+ ZA-B (20 μM) | 69.211 ± 5.204 | 1.5 | 1310.133 ± 1.797 * | 2.9 |
+ ZA-C (10 μM) | 99.561 ± 8.025 | 1.0 | 2146.430 ± 129.070 | 1.8 |
+ ZA-C (20 μM) | 83.740 ± 2.555 | 1.2 | 621.683 ± 17.651 * | 6.1 |
Cell Line | HeLaS3 | KB/VIN | ||
Compound | IC50 ± S.E. (nM) | RF | IC50 ± S.E. (nM) | RF |
Paclitaxel | 4.994 ± 0.077 | 1.0 | 510.512 ± 11.020 | 1.0 |
+ ZA-A (10 μM) | 1.509 ± 0.032 * | 3.3 | 141.745 ± 0.293 * | 3.6 |
+ ZA-A (20 μM) | 0.793 ± 0.028 * | 6.3 | 45.807 ± 0.128 * | 11.1 |
+ ZA-B (10 μM) | 2.053 ± 0.002 * | 2.4 | 225.581 ± 1.992 * | 2.3 |
+ ZA-B (20 μM) | 0.609 ± 0.004 * | 8.2 | 139.363 ± 2.287 * | 3.7 |
+ ZA-C (10 μM) | 3.657 ± 0.031 | 1.4 | 247.282 ± 7.276 * | 2.1 |
+ ZA-C (20 μM) | 3.238 ± 0.146 | 1.5 | 215.503 ± 5.563 * | 2.4 |
Cell Line | HeLaS3 | KB/VIN | ||
Compound | IC50 ± S.E. (nM) | RF | IC50 ± S.E. (nM) | RF |
Vincristine | 17.326 ± 0.645 | 1.0 | 15730.719 ± 2402.023 | 1.0 |
+ ZA-A (10 μM) | 6.661 ± 0.820 * | 2.6 | 2007.168 ± 51.760 * | 7.8 |
+ ZA-A (20 μM) | 4.463 ± 0.549 * | 3.9 | 304.132 ± 28.753 * | 51.7 |
+ ZA-B (10 μM) | 8.838 ± 0.420 * | 2.0 | 1967.489 ± 98.547 * | 8.0 |
+ ZA-B (20 μM) | 6.590 ± 0.236 * | 2.6 | 1018.696 ± 138.704 * | 15.4 |
+ ZA-C (10 μM) | 17.004 ± 1.944 | 1.0 | 6952.094 ± 54.567 * | 2.3 |
+ ZA-C (20 μM) | 15.795 ± 0.928 | 1.1 | 2938.459 ± 874.146 * | 5.4 |
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Teng, Y.-N.; Wang, Y.-H.; Wu, T.-S.; Hung, H.-Y.; Hung, C.-C. Zhankuic Acids A, B and C from Taiwanofungus camphoratus Act as Cytotoxicity Enhancers by Regulating P-Glycoprotein in Multi-Drug Resistant Cancer Cells. Biomolecules 2019, 9, 759. https://doi.org/10.3390/biom9120759
Teng Y-N, Wang Y-H, Wu T-S, Hung H-Y, Hung C-C. Zhankuic Acids A, B and C from Taiwanofungus camphoratus Act as Cytotoxicity Enhancers by Regulating P-Glycoprotein in Multi-Drug Resistant Cancer Cells. Biomolecules. 2019; 9(12):759. https://doi.org/10.3390/biom9120759
Chicago/Turabian StyleTeng, Yu-Ning, Yen-Hsiang Wang, Tian-Shung Wu, Hsin-Yi Hung, and Chin-Chuan Hung. 2019. "Zhankuic Acids A, B and C from Taiwanofungus camphoratus Act as Cytotoxicity Enhancers by Regulating P-Glycoprotein in Multi-Drug Resistant Cancer Cells" Biomolecules 9, no. 12: 759. https://doi.org/10.3390/biom9120759