AKT serine/threonine protein kinase modulates bufalin-triggered intrinsic pathway of apoptosis in CAL 27 human oral cancer cells

Tsai,Shih-Chang; Lu,Chi-Cheng; Lee,Chao-Ying; Lin,Yung-Chang; Chung,Jing-Gung; Kuo,Sheng-Chu; Amagaya,Sakae; Chen,Fei-Na; Chen,Michael   Yuanchien; Chan,Shih-Feng; Yang,Jai-Sing

doi:10.3892/ijo.2012.1605

AKT serine/threonine protein kinase modulates bufalin-triggered intrinsic pathway of apoptosis in CAL 27 human oral cancer cells

Authors:
- Shih-Chang Tsai
- Chi-Cheng Lu
- Chao-Ying Lee
- Yung-Chang Lin
- Jing-Gung Chung
- Sheng-Chu Kuo
- Sakae Amagaya
- Fei-Na Chen
- Michael Yuanchien Chen
- Shih-Feng Chan
- Jai-Sing Yang
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Affiliations: Department of Biological Science and Technology, China Medical University, Taichung 404, Taiwan, R.O.C. , Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan, R.O.C. , School of Pharmacy, China Medical University, Taichung 404, Taiwan, R.O.C., Department of Veterinary Medicine, National Chung Hsing University, Taichung 402, Taiwan, R.O.C. , Graduate Institute of Pharmaceutical Chemistry, China Medical University, Taichung 404, Taiwan, R.O.C., Tsuzuki Institute for Traditional Medicine, China Medical University, Taichung 404, Taiwan, R.O.C., Department of Social Medicine, China Medical University, Taichung 404, Taiwan, R.O.C. , School of Dentistry, China Medical University, Taichung 404, Taiwan, R.O.C., Department of Pharmacology, China Medical University, Taichung 404, Taiwan, R.O.C.
Published online on: August 23, 2012 https://doi.org/10.3892/ijo.2012.1605
Pages: 1683-1692

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Abstract

Bufalin has been reported to induce apoptosis in a variety of cancers but little is demonstrated in oral squamous cell carcinoma (OSCC) cells. The present study investigated the inhibition of proliferation, cell cycle arrest and apoptotic effects of bufalin in CAL 27 human oral cancer cells. Bufalin inhibited the growth of CAL 27 cells in a concentration-dependent manner and an IC50 value of bufalin was about 125 nM for 24 h treatment using the MTT assay. Moreover, the cell cycle distribution was arrested at the G0/G1 phase in CAL 27 cells after bufalin exposure. Upon bufalin stimulation, the expression of Bcl-2 was significantly decreased while that of cytochrome c, Apaf-1 and AIF was increased compared to the control group by western blot analysis. An increase in the expression of the active form of caspases was found in bufalin-treated cells, and the caspase activities were also elevated. Bufalin-triggered apoptosis was blocked by specific inhibitors of caspase-9 (z-LEHD-fmk) and caspase-3 (z-DEVD-fmk), respectively. In contrast, CAL 27 cells overexpressing constitutively active AKT (CAL 27/CA-AKT) were exposed to bufalin at different concentrations, and cell growth remained unchanged. Bufalin exhibited minimal apoptotic effects on CAL 27/CA-AKT cells. Taken together, bufalin induced G0/G1 phase arrest and provoked the intrinsic apoptotic pathway via AKT activation in CAL 27 cells. Our data suggest that bufalin could be potentially efficacious in the treatment of oral cancer in the future.

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