Tetrandrine Enhances H2O2-Induced Apoptotic Cell Death Through Caspase-dependent Pathway in Human Keratinocytes.

Abstract

BACKGROUND:

Tetrandrine, a bis-benzylisoquinoline alkaloid, induces apoptosis of many types of human cancer cell. Hydrogen peroxide (H2O2) is a reactive oxygen species inducer; however, there are no reports to show whether pre-treatment of tetrandrine with H2O2 induces more cell apoptosis than H2O2 alone. Thus, the present study investigated the effects of tetrandrine on H2O2-induced cell apoptosis of human keratinocytes, HaCaT, in vitro.

MATERIALS AND METHODS:

HaCaT cells were pre-treated with and without tetrandrine for 1 h, and then treated with H2O2 for examining cell morphological changes and cell viability using contrast-phase microscopy and propidium iodide (PI) exclusion assay, respectively. Cells were measured apoptotic cell death by using annexin V/PI double staining and further analyzed by flow cytometer. Cells were further assessed for DNA condensation using 2-(4-amidinophenyl)-6-indolecarbamidine staining. Western blotting was used to measure expression of apoptosis-associated proteins and confocal laser microscopy was used to measure the protein expression and nuclear translocation from the cytoplasm to nuclei.

RESULTS:

Pre-treatment of tetrandrine for 1 h and treatment with H2O2 enhanced H2O2-induced cell morphological changes and reduced cell viability, whilst increasing apoptotic cell death and DNA condensation. Furthermore, tetrandrine significantly increased expression of reactive oxygen species-associated proteins such as superoxide dismutase (Cu/Zn) and superoxide dismutase (Mn) but significantly reduced the level of catalase, which was also confirmed by confocal laser microscopy. It also increased expression of DNA repair-associated proteins ataxia telangiectasia mutated, ataxia-telangectasia and Rad3-related, phospho-P53, P53 and phosphorylated histone H2AX, and of pro-apoptotic proteins BCL2 apoptosis regulator-associated X-protein, caspase-3, caspase-8, caspase-9 and poly ADP ribose polymerase in HaCaT cells.

CONCLUSION:

These are the first and novel findings showing tetrandrine enhances H2O2-induced apoptotic cell death of HaCaT cells and may provide a potent approach for the treatment of proliferated malignant keratinocytes.

• Authors: Yi-Ching Cheng, Chao-Lin Kuo, Sheng-Yao Hsu, Tzong-DER Way, Ching-Ling Cheng, Jaw-Chyun Chen, Kuo-Ching Liu, Shu-Fen Peng, Wai-Jane Ho, Fu-Shin Chueh, Wen-Wen Huang
• Journal: In vivo (Athens, Greece) (In Vivo) 2021 Jul-Aug Vol. 35 Issue 4 Pg. 2047-2057 ISSN: 1791-7549 [Electronic] Greece
• PMID: 34182480 (Publication Type: Journal Article)
• Copyright: Copyright © 2021 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

Chemical References

• Benzylisoquinolines
• Reactive Oxygen Species
• tetrandrine
• Hydrogen Peroxide
• Caspases

Topics

• Apoptosis
• Benzylisoquinolines (pharmacology)
• Caspases (genetics)
• Cell Survival
• Humans
• Hydrogen Peroxide
• Keratinocytes
• Reactive Oxygen Species