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Anti-proliferative activity of biochanin A in human osteosarcoma cells via mitochondrial-involved apoptosis

Hsu, Yen-Nien, Shyu, Huey-Wen, Hu, Tsui-Wen, Yeh, Jou-Pei, Lin, Ya-Wen, Lee, Ling-Yi, Yeh, Yao-Tsung, Dai, Hong-Ying, Perng, Daw-Shyong, Su, Shu-Hui, Huang, Yu-Hsuan, Su, Shu-Jem
Food and chemical toxicology 2018 v.112 pp. 194-204
Trifolium pratense, acetylcysteine, antioxidants, apoptosis, biochanin A, caspase-3, caspase-9, cell growth, chemoprevention, cytochrome c, cytotoxicity, dose response, doxorubicin, glutathione, humans, membrane potential, mitochondria, mitochondrial membrane, mitogen-activated protein kinase, osteosarcoma, phosphatidylserines, toxicology, vitamin E
Biochanin A is a major isoflavone in red clover and a potent chemopreventive agent against cancer. However, the effects of biochanin A on human osteosarcoma cells have never been clarified. This study investigated the anti-proliferative potential of biochanin A in osteosarcoma cells. The results indicate that biochanin A inhibited cell growth and colony formation in a dose-dependent manner with a minimal toxicity to normal cells. The combination of doxorubicin and biochanin A could synergistically inhibit osteosarcoma cell growth. The cytotoxic effect of biochanin A via the induction of apoptosis as evidenced by formation of apoptotic bodies, externalization of phosphatidylserine, accumulation of sub-G1 phase cells, caspase 3 activation, and cleavage of PARP. Apoptosis was associated with loss of the mitochondrial membrane potential, release of cytochrome c, caspase 9 activation, increased Bax expression, and reduced Bcl-2 and Bcl-XL expression. Pre-treatment with a caspase-9 specific inhibitor (Z-LEHD-FMK) partially attenuated cell death, suggesting involvement of the intrinsic mitochondrial apoptotic cascade. However, pre-treatment with the JNK inhibitor SP600125, the MEK inhibitor PD-98059, and the p38 MAPK inhibitor SB203580 or the antioxidants vitamin E, N-acetylcysteine, and glutathione failed to prevent biochanin A-induced cell death. Our results suggest that biochanin A inhibits cell growth and induces apoptosis in osteosarcoma cells by triggering activation of the intrinsic mitochondrial pathway and caspase-9 and -3 and increasing the Bax: Bcl-2/Bcl-XL ratio.