Abstract
Natural flavonoids from plants have been demonstrated to possess promising chemopreventive activities against various diseases. 7-{4-[Bis-(2-hydroxy-ethyl)-amino]-butoxy}-5-hydroxy-8-methoxy-2-phenyl-chromen-4-one (V8), a newly synthesized derivative of wogonin may have antioxidant, antiviral, anti-inflammatory and anti-tumor potentials as wogonin. Based on the recent findings of V8, the anti-tumor activities and fundamental mechanisms by which V8 inhibits growth of hepatocellular carcinoma were further investigated in this study. After the treatment of V8, a significant inhibition of HepG2 cell proliferation was observed in a dose-dependent manner with the IC50 value of 23 μM using MTT assay. The exposure to V8 also resulted in apoptosis induction and an accumulation of ROS and Ca2+. Meanwhile, a release of cytochrome c (Cyt-c), activation of BH-3 only proteins and Bax, decrease in mitochondrial membrane potential ΔΨ, as well as a suppression of Bcl-2, pro-caspase9 and pro-caspase3 expression were shown. Moreover, knocking down CHOP partly decreased the effect of V8-mediated apoptosis and activation of GRP78, p-PERK, p-eIF2α, ATF4 and CHOP modulated ER stress triggered by V8. In vivo, V8 inhibited the transplanted mice H22 liver carcinomas in a dose-dependent manner. Compared with wogonin, V8 exhibited stronger anti-proliferative effects both in vitro and in vivo. The underlying mechanism of activating PERK-eIF2α-ATF4 pathway by which V8 induces apoptosis was verified once again in vivo. The apoptosis induction via the mitochondrial pathway by modulating the ROS-mediated ER signaling pathway might serve to provide support for further studies of V8 as a possible anticancer drug in the clinical treatment of cancer.
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Abbreviations
- ROS:
-
Reactive oxygen species
- ER:
-
Endoplasmic reticulum
- HCC:
-
Hepatocellular carcinoma
- UPR:
-
Unfolded protein response
- ΔΨm:
-
Mitochondrial transmembrane potential
- CHOP:
-
CCAAT/enhancer-binding protein–homologous protein
- DMSO:
-
Dimethyl sulfoxide
- NAC:
-
N-acetyl cysteine
- carboxy-H2DCFDA:
-
Dichlorofluorescein diacetate
- ERAD:
-
ER-associated degradation
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Acknowledgments
This work was supported by Program for Changjiang Scholars and Innovative Research Team in University (IRT1193), the Project Program of State Key Laboratory of Natural Medicines, China Pharmaceutical University(No. JKGZ201101), the National Science & Technology Major Project (No.2012ZX09304-001) and Natural Science Foundation of Jiangsu province (No.BK2009297 and No.BK2010432).
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Yi Zhang and Li Zhao have contributed equally to this article.
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Zhang, Y., Zhao, L., Li, X. et al. V8, a newly synthetic flavonoid, induces apoptosis through ROS-mediated ER stress pathway in hepatocellular carcinoma. Arch Toxicol 88, 97–107 (2014). https://doi.org/10.1007/s00204-013-1085-6
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DOI: https://doi.org/10.1007/s00204-013-1085-6