Combined Treatment with Low Cytotoxic Ethyl Acetate Nepenthes Extract and Ultraviolet-C Improves Antiproliferation to Oral Cancer Cells via Oxidative Stress
Abstract
:1. Introduction
2. Materials and Methods
2.1. Crude Extracts and Chemical
2.2. Cell Cultures and Proliferation Determination
2.3. UVC Irradiation and EANA Treatment
2.4. Cell Cycle Analysis
2.5. Apoptosis
2.6. Intracellular ROS
2.7. Mitochondrial Superoxide (MitoSOX)
2.8. Mitochondrial Membrane Potential (MitoMP)
2.9. Antioxidant Gene Expressions
2.10. γH2AX
2.11. 8-Hydroxy-2′-Deoxyguanosine (8-OxodG)
2.12. Statistics
3. Results
3.1. UVC Sensitizing Responses of EANA in Oral Cancer Cells
3.2. Cell Cycle Analysis of UVC and/or EANA Treatments in Oral Cancer Cells
3.3. Annexin V/7AAD Changes of UVC and/or EANA Treatments in Oral Cancer Cells
3.4. Caspase Activation of UVC and/or EANA Treatments in Oral Cancer Cells
3.5. ROS Changes of UVC and/or EANA Treatments in Oral Cancer Cells
3.6. MitoSOX Changes of UVC and/or EANA Treatments in Oral Cancer Cells
3.7. MitoMP Changes of UVC and/or EANA Treatments in Oral Cancer Cells
3.8. Antioxidant Gene Expressions of UVC and/or EANA Treatments in Oral Cancer Cells
3.9. γH2AX Changes of UVC and/or EANA Treatments in Oral Cancer Cells
3.10. 8-oxodG Changes of UVC and/or EANA Treatments in Oral Cancer Cells
4. Discussion
4.1. Radiosensitizers with Low Side Effects Are Helpful in Cancer Therapy
4.2. Oxidative Stresses Are Synergistically Increased in the Combined Treatment (UVC/EANA) of Oral Cancer Cells
4.3. DNA Damages Are Synergistically Increased in the Combined Treatment (UVC/EANA) of Oral Cancer Cells
4.4. Apoptosis Is Synergistically Increased in the Combined Treatment (UVC/EANA) of Oral Cancer Cells
4.5. Cell Cycle Arresting at G2/M Is Synergistically Increased in the Combined Treatment (UVC/EANA) of Oral Cancer Cells
4.6. All Separate and Synergistic Changes in UVC and/or EANA of Oral Cancer Cells Are Oxidative Stress Dependent
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Genes | Forward Primers (5′→3′) | Reverse Primers (5′→3′) | Length |
---|---|---|---|
RELA | TCAAGATCTGCCGAGTGAAC [33] | GACGATCGTCTGTATCTGGC | 306 bp |
FOXO3 | TACGAGTGGATGGTGCGTT | GGCTTTTCCGCTCTTCC | 189 bp |
SOD1 | AGGGCATCATCAATTTCGAGC [36] | CCCAAGTCTCCAACATGCCTC [37] | 211 bp |
HMOX1 | CCTTCTTCACCTTCCCCAACAT [37] | GGCAGAATCTTGCACTTTGTTGC [37] | 251 bp |
GAPDH | CCTCAACTACATGGTTTACATGTTCC [38] | CAAATGAGCCCCAGCCTTCT [39] | 220 bp |
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Peng, S.-Y.; Lin, L.-C.; Yang, Z.-W.; Chang, F.-R.; Cheng, Y.-B.; Tang, J.-Y.; Chang, H.-W. Combined Treatment with Low Cytotoxic Ethyl Acetate Nepenthes Extract and Ultraviolet-C Improves Antiproliferation to Oral Cancer Cells via Oxidative Stress. Antioxidants 2020, 9, 876. https://doi.org/10.3390/antiox9090876
Peng S-Y, Lin L-C, Yang Z-W, Chang F-R, Cheng Y-B, Tang J-Y, Chang H-W. Combined Treatment with Low Cytotoxic Ethyl Acetate Nepenthes Extract and Ultraviolet-C Improves Antiproliferation to Oral Cancer Cells via Oxidative Stress. Antioxidants. 2020; 9(9):876. https://doi.org/10.3390/antiox9090876
Chicago/Turabian StylePeng, Sheng-Yao, Li-Ching Lin, Zhe-Wei Yang, Fang-Rong Chang, Yuan-Bin Cheng, Jen-Yang Tang, and Hsueh-Wei Chang. 2020. "Combined Treatment with Low Cytotoxic Ethyl Acetate Nepenthes Extract and Ultraviolet-C Improves Antiproliferation to Oral Cancer Cells via Oxidative Stress" Antioxidants 9, no. 9: 876. https://doi.org/10.3390/antiox9090876