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Daphnegiravone D synergistically inhibits liver cancer in combination with oxaliplatin in vitro and in vivo
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Yu-Jue Wang1, 2, 3, 4, Jiao-Yang Hou1, 2, 3, 4, Xin-Ye Wang1, 2, 3, 4, Hong-Fei Chen1, 2, 3, 4, Shuang Qiu1, 2, 3, 4, Qing-Bo Liu1, 2, 3, 4, Xiao-Xiao Huang1, 2, 3, 4, Guo-Dong Yao1, 2, 3, 4, *, Shao-Jiang Song1, 2, 3, 4, *
Acupuncture and Herbal Medicine | 2026, 6(3) : 372 - 386
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Acupuncture and Herbal Medicine | 2026, 6(3): 372-386
• Original Articles •
Daphnegiravone D synergistically inhibits liver cancer in combination with oxaliplatin in vitro and in vivo
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Yu-Jue Wang1, 2, 3, 4, Jiao-Yang Hou1, 2, 3, 4, Xin-Ye Wang1, 2, 3, 4, Hong-Fei Chen1, 2, 3, 4, Shuang Qiu1, 2, 3, 4, Qing-Bo Liu1, 2, 3, 4, Xiao-Xiao Huang1, 2, 3, 4, Guo-Dong Yao1, 2, 3, 4, *, Shao-Jiang Song1, 2, 3, 4, *
Affiliations
  • 1Key Laboratory of Computational Chemistry-Based Natural Antitumor Drug Research & Development, Shenyang Pharmaceutical University, Shenyang, China
  • 2School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang, China
  • 3Engineering Research Center of Natural Medicine Active Molecule Research & Development, Liaoning, Shenyang Pharmaceutical University, Shenyang, China
  • 4Key Laboratory of Natural Bioactive Compounds Discovery & Modification, Shenyang, Shenyang Pharmaceutical University, Shenyang, China
About Author:

Yu-Jue Wang and Jiao-Yang Hou contributed equally to this work.

Published: 2026-09-25 doi: 10.1097/HM9.0000000000000201
Outline
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Objective

Daphnegiravone D (DGD), a prenylated flavonoid from Daphne giraldii Nitsche, a plant whose roots and stem are used in Chinese medicine, exerts an inhibitory effect on liver cancer cells. The protein kinase ataxia telangiectasia-mutated and Rad3-related (ATR) is an important component of the DNA damage response, and its inhibition enhances the sensitivity of some cancer cells to DNA-damaging drugs. Oxaliplatin (OXA) is a third-generation, platinum-based anticancer drug that exerts its inhibitory effects on liver cancer cells by inhibiting DNA synthesis. Owing to the fact that most patients develop drug resistance, finding novel treatments for liver cancer is urgent. In a previous study, DGD was shown to exert anti-tumor effects as a potential ATR kinase inhibitor. This study investigated the mechanism of action of DGD and its combination with OXA in liver cancer.

Methods

HepG2 and Hep3B cells were used to evaluate the inhibitory effects of DGD on liver cancer. Cellular thermal shift assay, co-immunoprecipitation, and western blotting were used to confirm the impact of DGD on DNA damage. The methyl thiazolyl tetrazolium (MTT) assay was performed, and the resulting data were used to calculate drug synergy scores. Acridine orange/ethidium bromide staining, Annexin V/propidium iodide staining, reactive oxygen species–related staining, and western blotting were performed to assess the efficacy of the combination of DGD and OXA. An in vivo tumor xenograft model in nude mice was used to investigate the efficacy of this combination. Hematoxylin–eosin staining and immunohistochemistry were performed to observe the condition of tissues in vivo.

Results

DGD inhibited liver cancer cells by affecting the formation of the ATR-ATRIP complex and downregulating the activator protein TopBP1. We found that DGD and OXA showed synergistic growth-inhibitory effects in liver cancer cells in vitro. DGD and OXA showed synergistic growth-inhibitory effects in liver cancer cells in vitro and showed lower cytotoxicity toward L-02 cells than an equi-effective higher concentration of OXA under the tested conditions in normal liver cells. The combination was associated with enhanced apoptosis, mitochondrial dysfunction, and oxidative stress. Further experiments suggested that DGD may enhance OXA-induced DNA damage, at least partly in association with suppression of ATR-related signaling. Consistent with the above results, DGD enhanced the antitumor efficacy of OXA in vivo without causing overt additional toxicity under the present experimental conditions.

Conclusion

Our findings indicate that DGD enhances the antitumor activity of OXA against liver cancer, with the in vitro synergistic effect being associated, at least partly, with suppression of ATR-related signaling, enhanced DNA damage, and oxidative stress. DGD also improved the antitumor efficacy of OXA in the xenograft model without overt additional toxicity.

ATR  /  Daphnegiravone D  /  Drug combination  /  Liver cancer  /  Oxaliplatin
Yu-Jue Wang, Jiao-Yang Hou, Xin-Ye Wang, Hong-Fei Chen, Shuang Qiu, Qing-Bo Liu, Xiao-Xiao Huang, Guo-Dong Yao, Shao-Jiang Song. Daphnegiravone D synergistically inhibits liver cancer in combination with oxaliplatin in vitro and in vivo[J]. Acupuncture and Herbal Medicine, 2026 , 6 (3) : 372 -386 . DOI: 10.1097/HM9.0000000000000201
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Funding
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  • National Natural Science Foundation of China(81973528; 82273821)
  • LiaoNing Revitalization Talents Program(XLYC2503156)
  • Youth Innovation Team Scientific Research Project of Liaoning Provincial Department of Education(LT222410163052)
  • Natural Science Foundation of Liaoning province(2022-MS-244)
  • Young Elite Scientists Sponsorship Program by CACM(2023-QNRC2-B24)
  • Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-D-202208)
  • Plan of Shenyang Young and Middle-aged Science and Technology Innovation Talents Support(RC210451)
  • Scientific Research Fund Project of Liaoning Provincial Department of Education(LJKZ0907)
  • China Postdoctoral Science Foundation(2019M661135)
  • Song Shaojiang Expert Workstation of Yunnan Province(202305 AF150030)
  • Outstanding and Excellent Youth Talent Support Program of Shenyang Pharmaceutical University(ZQ202201; YQ202104)
  • Liaoning Revitalization Talents Program(XLYC2002066)
  • Science and Technology Planning Project of Liaoning Province(2021JH1/10400049)
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Year 2026 volume 6 Issue 3
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Article Info
doi: 10.1097/HM9.0000000000000201
  • Receive Date：2025-11-22
  • Online Date：2026-11-04
  • Published：2026-09-25
Article Data
Affiliations
History
  • Received：2025-11-22
  • Accepted：2026-04-26
Funding
National Natural Science Foundation of China(81973528; 82273821)
LiaoNing Revitalization Talents Program(XLYC2503156)
Youth Innovation Team Scientific Research Project of Liaoning Provincial Department of Education(LT222410163052)
Natural Science Foundation of Liaoning province(2022-MS-244)
Young Elite Scientists Sponsorship Program by CACM(2023-QNRC2-B24)
Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-D-202208)
Plan of Shenyang Young and Middle-aged Science and Technology Innovation Talents Support(RC210451)
Scientific Research Fund Project of Liaoning Provincial Department of Education(LJKZ0907)
China Postdoctoral Science Foundation(2019M661135)
Song Shaojiang Expert Workstation of Yunnan Province(202305 AF150030)
Outstanding and Excellent Youth Talent Support Program of Shenyang Pharmaceutical University(ZQ202201; YQ202104)
Liaoning Revitalization Talents Program(XLYC2002066)
Science and Technology Planning Project of Liaoning Province(2021JH1/10400049)
Affiliations
    1Key Laboratory of Computational Chemistry-Based Natural Antitumor Drug Research & Development, Shenyang Pharmaceutical University, Shenyang, China
    2School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang, China
    3Engineering Research Center of Natural Medicine Active Molecule Research & Development, Liaoning, Shenyang Pharmaceutical University, Shenyang, China
    4Key Laboratory of Natural Bioactive Compounds Discovery & Modification, Shenyang, Shenyang Pharmaceutical University, Shenyang, China

Corresponding:

* Guo-Dong Yao, E-mail: ;
Shao-Jiang Song, E-mail: .
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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