Abstract
Escin, a triterpene saponin isolated from horse chestnut seed, has been used to treat encephaledema, tissue swelling and chronic venous insufficiency. Recent studies show that escin induces cell cycle arrest, tumor proliferation inhibition and tumor cell apoptosis. But the relationship between escin-induced DNA damage and cell apoptosis in tumor cells remains unclear. In this study, we investigated whether and how escin-induced DNA damage contributed to escin-induced apoptosis in human colorectal cancer cells. Escin (5–80 μg/mL) dose-dependently inhibited the cell viability and colony formation in HCT116 and HCT8 cells. Escin treatment induced DNA damage, leading to p-ATM and γH2AX upregulation. Meanwhile, escin treatment increased the expression of p62, an adaptor protein, which played a crucial role in controlling cell survival and tumorigenesis, and had a protective effect against escin-induced DNA damage: knockdown of p62 apparently enhanced escin-induced DNA damage, whereas overexpression of p62 reduced escin-induced DNA damage. In addition, escin treatment induced concentration- and time-dependent apoptosis. Similarly, knockdown of p62 significantly increased escin-induced apoptosis in vitro and produced en escin-like antitumor effect in vivo. Overexpression of p62 decreased the rate of apoptosis. Further studies revealed that the functions of p62 in escin-induced DNA damage were associated with escin-induced apoptosis, and p62 knockdown combined with the ATM inhibitor KU55933 augmented escin-induced DNA damage and further increased escin-induced apoptosis. In conclusion, our results demonstrate that p62 regulates ATM/γH2AX pathway-mediated escin-induced DNA damage and apoptosis.
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Acknowledgements
This work was partially supported by the National Natural Science Foundation of China (No 81672970), the Natural Science Foundation of Jiangsu Province (No BK20160338), and supported by Suzhou Key Medical Center (No LCZX201505), the Projects of Suzhou Technology Bureau (No SS201753, SYS201552, SZS201618), the Second Affiliated Hospital of Soochow University Preponderant Clinic Discipline Group Project Funding, the Project of Invigorating Health Care through Science, Technology and Education (Jiangsu Provincial Medical Youth Talent, QNRC2016249) and Graduate Student Scientific Research Innovation Projects of Jiangsu Province (No KYCX17_1999), National Natural Science Youth Foundation of China (No 81602613).
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Wang, Z., Chen, Q., Li, B. et al. Escin-induced DNA damage promotes escin-induced apoptosis in human colorectal cancer cells via p62 regulation of the ATM/γH2AX pathway. Acta Pharmacol Sin 39, 1645–1660 (2018). https://doi.org/10.1038/aps.2017.192
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DOI: https://doi.org/10.1038/aps.2017.192
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