Riceberry Rice Bran Protein Hydrolyzed Fractions Induced Apoptosis, Senescence and G1/S Cell Cycle Arrest in Human Colon Cancer Cell Lines
Abstract
:1. Introduction
2. Materials and Methods
2.1. Riceberry Rice Bran Hydrolyzation, Extraction, Anthocyanin Removal, and Fractionation
2.2. Cell Culture
2.3. Cellular Viability by MTT Assay
2.4. Senescence-Associated Beta-Galactosidase (SA-β-gal) Staining
2.5. Acridine Orange/Propidium Iodide (AO/PI) Double Staining Test
2.6. Apoptosis Assay
2.7. Cell Cycle Analysis
2.8. Statical Analysis
3. Results
3.1. Cytotoxicity of Hydrolyzed Riceberry Rice Bran Extract on Colon Cancer Cell Lines
3.2. Senescence Inductive Effect of Hydrolyzed Riceberry Rice Bran Extract on Colon Cancer Cell Lines
3.3. Apoptotic Inductive Effect of Hydrolyzed Riceberry Rice Bran Extract on Colon Cancer Cell Lines
3.4. The Analysis of Cell Apoptosis by Annexin V and Flow Cytometry
3.5. Cell Cycle Analysis by Flow Cytometry
3.6. Cytotoxicity of HRBE Fractions on Colon Cancer Cell Lines
3.7. Apoptotic Inductive Effect of the >50 kDa HRBE Fraction on Metastatic Colon Cancer Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Time/Treatment | IC50 (µg/mL) | |||||
---|---|---|---|---|---|---|
HT-29 | SW-620 | PCS-201-010 | ||||
HRBE | Mitomycin C | HRBE | Mitomycin C | HRBE | Mitomycin C | |
24 h | 43,677.08 ± 30,191.29 | 762.91 ± 650.87 | 9879.66 ± 1855.29 | 192.93 ± 90.21 | 15,351.45 ± 7954.70 | 209.78 ± 78.02 |
48 h | 11,164.35 ± 1642.52 | 141.26 ± 59.35 | 6869.84 ± 1674.36 | 57.60 ± 25.79 | 8006.75 ± 1077.60 | 177.10 ± 65.48 |
72 h | 6053.50 ± 605.86 | 56.99 ± 2.88 | 5467.95 ± 1143.94 | 39.65 ± 12.15 | 6745.09 ± 1016.33 | 145.02 ± 23.67 |
The Percentage Yield of Extracts | ||
---|---|---|
Fraction size | >50 kDa | 28.74 |
50–30 kDa | 15.25 | |
30–10 kDa | 19.4 | |
10–3 kDa | 12.29 | |
<3 kDa | 10.67 |
Time/Treatment | IC50 | ||||||
---|---|---|---|---|---|---|---|
The Fraction of HRBE (µg/mL) | Doxorubicin (µg/mL) | Mitomycin C (µg/mL) | |||||
>50 kDa | 50–30 kDa | 30–10 kDa | 10–3 kDa | <3 kDa | |||
24 h | 27,457.44 ± 8871.23 | 46,314.12± 42,427.4 | 30,242.36± 3395.62 | 106,535.55 ± 32,830.31 | 90,793.65 ± 5005.19 | 273.72 ± 0.1 | 33.34 ± 4.67 |
48 h | 11,209.20 ± 3219.13 | 39,100.08 ± 22,822.23 | 13,679.24 ± 4151.03 | 28,289.96 ± 15,349.07 | 18,644.05 ± 1741.60 | 91.89 ± 0.12 | 12.77 ± 1.84 |
72 h | 4,864.93 ± 1187.25 | 8669.95 ± 2437.84 | 290,377.02 ± 47,713.73 | 13,672.91 ± 4561.09 | 11,357.97 ± 2577.29 | 77.24 ± 0.09 | 3.5 ± 7.79 |
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Wattayagorn, V.; Kongsema, M.; Tadakittisarn, S.; Chumnanpuen, P. Riceberry Rice Bran Protein Hydrolyzed Fractions Induced Apoptosis, Senescence and G1/S Cell Cycle Arrest in Human Colon Cancer Cell Lines. Appl. Sci. 2022, 12, 6917. https://doi.org/10.3390/app12146917
Wattayagorn V, Kongsema M, Tadakittisarn S, Chumnanpuen P. Riceberry Rice Bran Protein Hydrolyzed Fractions Induced Apoptosis, Senescence and G1/S Cell Cycle Arrest in Human Colon Cancer Cell Lines. Applied Sciences. 2022; 12(14):6917. https://doi.org/10.3390/app12146917
Chicago/Turabian StyleWattayagorn, Vichugorn, Mesayamas Kongsema, Sukuntaros Tadakittisarn, and Pramote Chumnanpuen. 2022. "Riceberry Rice Bran Protein Hydrolyzed Fractions Induced Apoptosis, Senescence and G1/S Cell Cycle Arrest in Human Colon Cancer Cell Lines" Applied Sciences 12, no. 14: 6917. https://doi.org/10.3390/app12146917
APA StyleWattayagorn, V., Kongsema, M., Tadakittisarn, S., & Chumnanpuen, P. (2022). Riceberry Rice Bran Protein Hydrolyzed Fractions Induced Apoptosis, Senescence and G1/S Cell Cycle Arrest in Human Colon Cancer Cell Lines. Applied Sciences, 12(14), 6917. https://doi.org/10.3390/app12146917