Kaempferol induces autophagy through AMPK and AKT signaling molecules and causes G2/M arrest via downregulation of CDK1/cyclin B in SK-HEP-1 human hepatic cancer cells

Huang,Wen-Wen; Tsai,Shih-Chang; Peng,Shu-Fen; Lin,Meng-Wei; Chiang,Jo-Hua; Chiu,Yu-Jen; Fushiya,Shinji; Tseng,Michael  T.; Yang,Jai-Sing

doi:10.3892/ijo.2013.1909

Kaempferol induces autophagy through AMPK and AKT signaling molecules and causes G2/M arrest via downregulation of CDK1/cyclin B in SK-HEP-1 human hepatic cancer cells

Authors:
- Wen-Wen Huang
- Shih-Chang Tsai
- Shu-Fen Peng
- Meng-Wei Lin
- Jo-Hua Chiang
- Yu-Jen Chiu
- Shinji Fushiya
- Michael T. Tseng
- Jai-Sing Yang
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Affiliations: Department of Biological Science and Technology, China Medical University, Taichung 40402, Taiwan, R.O.C., Department of Nursing, Cardinal Tien College of Healthcare and Management, Taipei 23143, Taiwan, R.O.C., Department of Life Sciences, National Chung Hsing University, Taichung 40227, Taiwan, R.O.C., Department of Surgery, Taipei Veterans General Hospital, Taipei 11217, Taiwan, R.O.C., Department of Kampo Pharmaceutical Sciences, Nihon Pharmaceutical University, Saitama 362-0806, Japan, Department of Anatomical Sciences and Neurobiology, School of Medicine, University of Louisville, Louisville, KY 40202, USA, Department of Pharmacology, China Medical University, Taichung 40402, Taiwan, R.O.C.
Published online on: April 17, 2013 https://doi.org/10.3892/ijo.2013.1909
Pages: 2069-2077

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Abstract

Kaempferol belongs to the flavonoid family and has been used in traditional folk medicine. Here, we investigated the antitumor effects of kaempferol on cell cycle arrest and autophagic cell death in SK-HEP-1 human hepatic cancer cells. Kaempferol decreased cell viability as determined by MTT assays and induced a G2/M phase cell cycle arrest in a concentration-dependent manner. Kaempferol did not induce DNA fragmentation, apoptotic bodies or caspase-3 activity in SK-HEP-1 cells as determined by DNA gel electrophoresis, DAPI staining and caspase-3 activity assays, respectively. In contrast, kaempferol is involved in the autophagic process. Double-membrane vacuoles, lysosomal compartments, acidic vesicular organelles and cleavage of microtubule-associated protein 1 light chain 3 (LC3) were observed by transmission electron microscopy, LysoΤracker red staining, GFP-fluorescent LC3 assays and acridine orange staining, respectively. In SK-HEP-1 cells, kaempferol increased the protein levels of p-AMPK, LC3-II, Atg 5, Atg 7, Atg 12 and beclin 1 as well as inhibited the protein levels of CDK1, cyclin B, p-AKT and p-mTOR. Taken together, CDK1/cyclin B expression and the AMPK and AKT signaling pathways contributed to kaempferol-induced G2/M cell cycle arrest and autophagic cell death in SK-HEP-1 human hepatic cancer cells. These results suggest that kaempferol may be useful for long-term cancer prevention.

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