Activation of the cyclic guanosine monophosphate-adenosine monophosphate adenosine synthetase-stimulator of interferon genes signaling pathway regulates the cytotoxicity of γδT cells against hepatoma cells

Shuai HU; Erqiang WANG; Xiaoyong DUO; Zhi XU; Yumeng ZHANG; Shiwei XIE; Lihua RONG; Yuchen WANG; Jiang LI; Shijie ZHANG

doi:10.12449/JCH251219

Volume 41 Issue 12

Dec. 2025

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Article Navigation > Journal of Clinical Hepatology > 2025 > 41(12): 2572-2580

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Activation of the cyclic guanosine monophosphate-adenosine monophosphate adenosine synthetase-stimulator of interferon genes signaling pathway regulates the cytotoxicity of γδT cells against hepatoma cells

DOI: 10.12449/JCH251219

Shuai HU¹,
Erqiang WANG³,
Xiaoyong DUO²,
Zhi XU¹,
Yumeng ZHANG¹,
Shiwei XIE¹,
Lihua RONG¹,
Yuchen WANG¹,
Jiang LI^{1, 4
,
,
,},
Shijie ZHANG^{1, 4
,
,
,}

1.
Department of Hepatobiliary Surgery，The First Affiliated Hospital of Shihezi University，Shihezi，Xinjiang 832000，China
2.
First Department of Critical Care Medicine，The First Affiliated Hospital of Shihezi University，Shihezi，Xinjiang 832000，China
3.
School of Basic Medical Sciences，Huazhong University of Science and Technology，Wuhan 430000，China
4.
NHC Key Laboratory of Prevention and Treatment of Central Asia High Incidence Diseases，Shihezi University，Shihezi，Xinjiang 832000，China

Research funding:

Open Research Fund of the Key Laboratory of Prevention and Treatment of High Incidence of Central Asia,NHC （National Health Commission） (KF202203);

Guided Science and Technology Plan Project of XPCC （Xinjiang Production and Construction Corps） (2023ZD023);

Hospital-Level Science and Technology Plan Project (BS202203)

More Information

Corresponding author: LI Jiang， jiangli08@163.com （ORCID： 0000-0003-0120-9239）; ZHANG Shijie， zhangshijie1@sina.com （ORCID： 0000-0002-2416-2421）
Received Date: 2025-04-12
Accepted Date: 2025-05-23
Published Date: 2025-12-25

Abstract

Abstract

Objective To investigate the regulatory effect of the cyclic guanosine monophosphate-adenosine monophosphate adenosine synthetase （cGAS）-stimulator of interferon genes （STING） signaling pathway on the cytotoxicity of γδT cells against hepatocellular carcinoma （HCC） through in vitro experiments， and to provide new ideas for improving the efficacy of adoptive immunotherapy based on γδT cells. Methods Peripheral blood mononuclear cells （PBMCs） were isolated and γδT cells were multiplied， and their purity was measured. Mature γδT cells were divided into γδT group， γδT-G10 group， and γδT-H-151 group. After 24 hours of in vitro stimulation， Western blot was used to measure the expression levels of key proteins in the cGAS-STING signaling pathway， and ELISA was used to measure the concentrations of interferon-γ （IFN-γ） and tumor necrosis factor-α （TNF-α）. The cells in each group were cocultured with MHCC-97H and Huh-7 cells for 6 hours， and then CCK8 assay was used to measure the survival rate of HCC cells in each group. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups， the Dunnett’s T3-test and the Tukey test were used for further comparision between two groups. Results Flow cytometry showed that the purity of γδT cells reached above 99%. Western blot showed that there was no significant difference in the expression of cGAS between the γδT group and the other two groups； compared with the γδT group， the γδT-G10 group had significant increases in the expression levels of STING， phosphorylated STING， TBK1， phosphorylated TBK1， interferon regulatory factor 3 （IRF3）， and phosphorylated IRF3， while the γδT-H-151 group had significant reductions in the expression of these proteins. ELISA showed that compared with the γδT group， the γδT-G10 group had significant increases in the secretion of IFN-γ and TNF-α by γδT cells （P<0.01 and P<0.05）， while the γδT-H-151 group had significant reductions in IFN-γ and TNF-α （P<0.01 and P<0.000 1）. CCK-8 assay showed that compared with the γδT group， the γδT-G10 group had significant reductions in the survival rates of the two HCC cell lines （P<0.000 1）， while the γδT-H-151 group showed significant increases （P<0.000 1）. Conclusion The cGAS-STING signaling pathway can regulate the cytotoxicity of γδT cells against HCC in vitro.
- Carcinoma， Hepatocellular,
- Guanylate Kinases,
- Adenosine Monophosphate,
- Interferon Regulatory Factor-1,
- T-Cell， gamma-delta

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Activation of the cyclic guanosine monophosphate-adenosine monophosphate adenosine synthetase-stimulator of interferon genes signaling pathway regulates the cytotoxicity of γδT cells against hepatoma cells

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Activation of the cyclic guanosine monophosphate-adenosine monophosphate adenosine synthetase-stimulator of interferon genes signaling pathway regulates the cytotoxicity of γδT cells against hepatoma cells

DOI: 10.12449/JCH251219

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