The Antiproliferative Effect of Chakasaponins I and II, Floratheasaponin A, and Epigallocatechin 3-O-Gallate Isolated from Camellia sinensis on Human Digestive Tract Carcinoma Cell Lines
Abstract
:1. Introduction
2. Results and Discussion
2.1. Antiproliferative Activities of Constituents Isolated from “Tea Flower” against Human Gastric Carcinoma HSC-2, HSC-4, MKN-45, and Caco-2 Cells
2.2. Effects of Cell Cycle Distribution in HSC-2 Cells
2.3. Quantification of Apoptotic Cell Death Using Annexin-V Binding Assay in HSC-2 Cells
2.4. Evaluation of Apoptotic Morphological Changes in HSC-2 Cells
2.5. DNA Fragmentation in HSC-2 Cells
2.6. Effects of Caspase-3/7 in HSC-2 Cells
3. Materials and Methods
3.1. Chemicals Constituents from “Tea Flower”
3.2. Reagents
3.3. Cell Viability Assays
3.4. Cell Cycle Analysis
3.5. Annexin-V/7-AAD Assay
3.6. DAPI Staining for Morphological Analysis
3.7. Agarose Gel Electrophoresis for the Detection of DNA Fragmentation
3.8. Caspase-3/7 Assay
3.9. Statistics
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Treatment | IC50 (µM) a | |||
---|---|---|---|---|
HSC-2 | HSC-4 | MKN-45 | Caco-2 | |
Chakasaponin I (1) | 4.6 | 17.5 | 16.8 | 30.2 |
Chakasaponin II (2) | 14.1 | 18.2 | 17.3 | 40.6 |
Floratheasaponin A (3) | 4.4 | 6.2 | 4.5 | 19.3 |
(–)-Epigallocatechin 3-O-gallate (4) | 28.3 | 27.2 | >100 (70.3) | >100 (71.8) |
5-FU | >100 (55.5) | >100 (77.3) | >100 (52.4) | >100 (80.2) |
Cisplatin | 14.5 | 7.0 | circa 100 | >100 (69.5) |
Doxorubicin | 0.040 | 0.18 | 0.12 | >100 (80.9) |
Camptothecin | 0.020 | 0.089 | 0.10 | >100 (63.7) |
Taxol | 0.0012 | 0.0059 | 0.15 | >100 (61.5) |
Treatment | Concentration (µM) | Cell Cycle (%) a | ||
---|---|---|---|---|
G0/G1 Phase | S Phase | G2/M Phase | ||
Control | – | 73.5 ± 0.8 | 10.8 ± 0.4 | 15.6 ± 0.7 |
Chakasaponin I (1) | 5 | 71.6 ± 0.4 | 10.2 ± 0.4 | 18.0 ± 0.1 |
– | 7.5 | 69.1 ± 1.4 * | 10.6 ± 0.8 | 20.1 ± 0.8 ** |
– | 10 | 59.9 ± 0.2 ** | 11.2 ± 1.0 | 28.7 ± 0.9 ** |
Control | – | 72.4 ± 1.1 | 11.6 ± 0.4 | 16.0 ± 0.7 |
Chakasaponin II (2) | 10 | 60.3 ± 0.1 ** | 10.5 ± 0.4 | 28.9 ± 0.3 ** |
– | 12.5 | 44.1 ± 1.3 ** | 18.8 ± 2.4 ** | 36.8 ± 1.3 ** |
– | 15 | 49.2 ± 0.8 ** | 20.0 ± 0.3 ** | 30.5 ± 0.6 ** |
Control | – | 76.5 ± 1.4 | 9.8 ± 0.6 | 13.5 ± 0.8 |
Floratheasaponin A (3) | 2.5 | 69.4 ± 1.0 * | 11.8 ± 0.1 | 18.5 ± 0.9 * |
– | 5 | 67.6 ± 1.7 ** | 10.3 ± 0.7 | 21.7 ± 0.9 ** |
– | 7.5 | 53.1 ± 0.8 ** | 15.9 ± 1.8 ** | 30.3 ± 1.1 ** |
Control | – | 78.5 ± 1.2 | 9.4 ± 0.5 | 11.8 ± 0.7 |
(–)-Epigallocatechin 3-O-gallate (4) | 25 | 78.8 ± 1.1 | 9.7 ± 0.5 | 11.2 ± 0.6 |
– | 50 | 78.6 ± 1.2 | 9.0 ± 0.5 | 12.0 ± 0.6 |
– | 75 | 76.8 ± 0.5 | 10.1 ± 0.1 | 12.7 ± 0.4 |
5-FU | 30 | 36.4 ± 0.3 ** | 37.9 ± 0.9 ** | 25.4 ± 0.6 ** |
Cisplatin | 3 | 50.7 ± 0.7 ** | 27.9 ± 0.5 ** | 20.0 ± 0.3 ** |
Treatment | Concentration (µM) | Total Apoptotic Cells (%) a |
---|---|---|
Control | – | 8.9 ± 1.5 |
Chakasaponin I (1) | 5 | 10.0 ± 0.4 |
– | 7.5 | 35.6 ± 3.0 ** |
– | 10 | 89.7 ± 0.4 ** |
Control | – | 6.3 ± 0.4 |
Chakasaponin II (2) | 10 | 7.0 ± 0.3 |
– | 12.5 | 35.7 ± 0.4 ** |
– | 15 | 64.2 ± 1.0 ** |
Control | – | 12.9 ± 0.4 |
Floratheasaponin A (3) | 2.5 | 17.5 ± 1.0 |
– | 5 | 18.7 ± 0.3 * |
– | 7.5 | 50.1 ± 1.9 ** |
Control | – | 13.9 ± 0.5 |
(–)-Epigallocatechin 3-O-gallate (4) | 25 | 14.4 ± 0.6 |
– | 50 | 13.1 ± 0.1 |
– | 75 | 12.7 ± 0.1 |
Camptothecin | 0.02 | 16.9 ± 0.2 ** |
– | 0.05 | 43.9 ± 0.7 ** |
Treatment | Concentration (µM) | Total Apoptotic Cells (%) a |
---|---|---|
Control | – | 7.1 ± 0.4 |
Chakasaponin I (1) | 5 | 25.9 ± 3.8 ** |
– | 7.5 | 59.6 ± 6.3 ** |
– | 10 | 90.2 ± 0.9 ** |
Control | – | 2.8 ± 0.1 |
Chakasaponin II (2) | 10 | 5.9 ± 0.1 |
– | 12.5 | 35.6 ± 1.5 ** |
– | 15 | 50.9 ± 9.7 ** |
Control | – | 4.2 ± 0.5 |
Floratheasaponin A (3) | 2.5 | 27.9 ± 0.6 ** |
– | 5 | 27.9 ± 1.0 ** |
– | 7.5 | 45.3 ± 0.6 ** |
Control | – | 7.2 ± 0.4 |
(–)-Epigallocatechin 3-O-gallate (4) | 25 | 5.9 ± 0.2 |
– | 50 | 4.9 ± 0.4 |
– | 75 | 5.9 ± 0.1 |
Camptothecin | 0.03 | 67.2 ± 1.2 ** |
– | 0.05 | 75.0 ± 2.1 ** |
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Kitagawa, N.; Morikawa, T.; Motai, C.; Ninomiya, K.; Okugawa, S.; Nishida, A.; Yoshikawa, M.; Muraoka, O. The Antiproliferative Effect of Chakasaponins I and II, Floratheasaponin A, and Epigallocatechin 3-O-Gallate Isolated from Camellia sinensis on Human Digestive Tract Carcinoma Cell Lines. Int. J. Mol. Sci. 2016, 17, 1979. https://doi.org/10.3390/ijms17121979
Kitagawa N, Morikawa T, Motai C, Ninomiya K, Okugawa S, Nishida A, Yoshikawa M, Muraoka O. The Antiproliferative Effect of Chakasaponins I and II, Floratheasaponin A, and Epigallocatechin 3-O-Gallate Isolated from Camellia sinensis on Human Digestive Tract Carcinoma Cell Lines. International Journal of Molecular Sciences. 2016; 17(12):1979. https://doi.org/10.3390/ijms17121979
Chicago/Turabian StyleKitagawa, Niichiro, Toshio Morikawa, Chiaki Motai, Kiyofumi Ninomiya, Shuhei Okugawa, Ayaka Nishida, Masayuki Yoshikawa, and Osamu Muraoka. 2016. "The Antiproliferative Effect of Chakasaponins I and II, Floratheasaponin A, and Epigallocatechin 3-O-Gallate Isolated from Camellia sinensis on Human Digestive Tract Carcinoma Cell Lines" International Journal of Molecular Sciences 17, no. 12: 1979. https://doi.org/10.3390/ijms17121979