Tetrandrine induces cell death in SAS human oral cancer cells through caspase activation-dependent apoptosis and LC3-I and LC3-II activation-dependent autophagy

Huang,An-Cheng; Lien,Jin-Cherng; Lin,Meng-Wei; Yang,Jai-Sing; Wu,Ping-Ping; Chang,Shu-Jen; Lai,Tung-Yuan

doi:10.3892/ijo.2013.1952

Tetrandrine induces cell death in SAS human oral cancer cells through caspase activation-dependent apoptosis and LC3-I and LC3-II activation-dependent autophagy

Authors:
- An-Cheng Huang
- Jin-Cherng Lien
- Meng-Wei Lin
- Jai-Sing Yang
- Ping-Ping Wu
- Shu-Jen Chang
- Tung-Yuan Lai
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Affiliations: Department of Nursing, St. Mary's Junior College of Medicine, Nursing and Management, Yilan, Taiwan, R.O.C., Graduate Institute of Pharmaceutical Chemistry, China Medical University, Taichung, Taiwan, R.O.C., Department of Nursing, Cardinal Tien Junior College of Healthcare and Management, Yilan, Taiwan, R.O.C., Department of Pharmacology, China Medical University, Taichung, Taiwan, R.O.C., School of Pharmacy, China Medical University, Taichung, Taiwan, R.O.C., Department of Traditional Medicine, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan, R.O.C.
Published online on: May 21, 2013 https://doi.org/10.3892/ijo.2013.1952
Pages: 485-494

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Abstract

Numerous studies have demonstrated that autophagy is associated with cancer development. Thus, agents to induce autophagy could be employed in some cases for the treatment of cancer. Our results showed that tetrandrine significantly decreased the viability of SAS cells in a concentration- and time-dependent manner. Tetrandrine induced nuclear condensation, demonstrated by DAPI staining. The early events in apoptosis analysed by Annexin V/PI staining indicated that the percentage of cells staining positive for Annexin V was slightly increased in SAS cells with tetrandrine treatment but was much lower following bafilomycin A1 pre-treatment. Tetrandrine caused AVO and MDC induction in SAS cells in a concentration-dependent manner by fluorescence microscopy. Tetrandrine also caused LC-3 expression in SAS cells in a time-dependent manner. Our results show that tetrandrine treatment induced the levels of cleaved caspase-3 in a concentration- and time-dependent manner. Tetrandrine treatment induced the levels of LC-3 II, Atg-5, beclin-1, p-S6, p-ULK, p-mTOR, p-Akt (S473) and raptor. Tetrandrine decreased cell viability, but bafilomycin A1, 3-MA, chloroquine and NAC protected tetrandrine-treated SAS cells against decrease of cell viability. Atg-5, beclin-1 siRNA decreased tetrandrine-induced cleaved caspase-3 and cleaved PARP in SAS cells and protected tetrandrine-treated SAS cells against decrease in cell viability. Chloroquine, NAC and bafilomycin A1 also decreased tetrandrine-induced cleaved caspase-3 and cleaved PARP in SAS cells. Our results indicate the tetrandrine induces apoptosis and autophagy of SAS human cancer cells via caspase-dependent and LC3-I and LC3-II‑dependent pathways.

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