三白草酮调控Krüppel样因子5-促红细胞生成素肝细胞A2通路对肝癌细胞生长和免疫逃逸的影响

doi:10.3969/j.issn.1006-5725.2025.10.008

Abstract

Abstract:

Objective To investigate the effects of sauchinone （Sch） on the growth and immune escape of liver cancer cells by regulating the Krüppel like factor 5 （KLF5）-erythropoietin-producing hepatocellular receptor A2 （EphA2） pathway. Methods Huh-7 cells were grouped into liver cancer group， Sch low， medium， and high-dose groups （Sch-L group， Sch-M group， Sch-H group）， Sch-H + KLF5 activator negative control group （OE-NC）， and Sch-H + KLF5 activator （OE-KLF5） group. 5-bromo-2-deoxyuracil （EdU） staining， CCK-8， scratch assay， and Transwll were used to detect the proliferation， migration， and invasion of Huh-7 cells， respectively. Western blot was applied to detect proliferative cell nuclear antigen （PCNA）， migration invasion enhancer（MIEN1）， matrix metalloproteinase-2 （MMP-2）， programmed death receptor ligand 1 （PD-L1）， KLF5， and EphA2 proteins in Huh-7 cells. The Huh-7 cells in above 6 groups were co cultured with activated CD8⁺ T cells in a 96 well plate and named as liver cancer co culture group， Sch-L co culture group， Sch-M co culture group， Sch-H co culture group， Sch-H + OE-NC co culture group， and Sch-H + OE-KLF5 co culture group. The killing rate of CD8⁺T cells in the co culture system and the levels of interferon-γ （IFN-γ）， interleukin-4 （IL-4）， and tumor necrosis factor-α （TNF-α） in the supernatant were detected. Results Compared with the liver cancer group， the EdU positive rate， OD₄₅₀ value， cell invasion number， and PCNA， MIEN1， MMP-2， PD-L1， KLF5， and EphA2 proteins in the Sch-L， Sch-M， and Sch-H groups reduced， the migration distance shorten （P < 0.05）. Compared with the Sch-H group and Sch-H + OE-NC group， the EdU positive rate， OD₄₅₀ value， cell invasion number， and PCNA， MIEN1， MMP-2， PD-L1， KLF5， and EphA2 proteins in the Sch-H + OE-KLF5 group increased， the migration distance prolonged （P < 0.05）. Compared with the liver cancer co culture group， the killing rate of CD8⁺T cells on Huh-7 cells and the levels of IFN-γ， TNF-α， and IL-4 in the supernatant in the Sch-L co culture group， Sch-M co culture group， and Sch-H co culture group increased （P < 0.05）. Compared with the Sch-H co culture group and the Sch-H + OE-NC co culture group， the killing rate of CD8⁺ T cells on Huh-7 cells and the levels of IFN-γ， TNF-α， and IL-4 in the supernatant in the Sch-H + OE-KLF5 co culture group decreased （P < 0.05）. Conclusion Sch may inhibit growth and immune escape of liver cancer cells by inhibiting the KLF5-EphA2 pathway.

Key words: sauchinone, liver cancer, immune escape, krüppel like factor 5-erythropoietin-producing hepatocellular receptor A2 pathway

CLC Number:

R285.5

Shan WANG,Jinliang WAN,Zhonghua ZHAO,Jing YANG,Jie ZHAO,Yanzhang. HAO. Effects of sauchinone regulating KLF5⁃EphA2 pathway on the growth and immune escape of hepatocellular carcinoma cells[J]. The Journal of Practical Medicine, 2025, 41(10): 1480-1486.

Figures/Tables 10

Fig.1

Tab.1

Fig.2

Tab.2

Fig.3

Tab.3

Tab.4

Tab.5

Fig.4

Tab.6

References 27

1	WANG R， LI R， YANG H， et al. Flavokawain C inhibits proliferation and migration of liver cancer cells through FAK/PI3K/AKT signaling pathway［J］. J Cancer Res Clin Oncol， 2024，150（3）：117-133. doi:10.1007/s00432-024-05639-z doi: 10.1007/s00432-024-05639-z
2	PARK H， LEE S， LEE J， et al. Exploring the JAK/STAT signaling pathway in hepatocellular carcinoma： Unraveling signaling complexity and therapeutic implications［J］. Int J Mol Sci， 2023，24（18）：13764-13786. doi:10.3390/ijms241813764 doi: 10.3390/ijms241813764
3	GUO F， YANG X， HU C， et al. Network pharmacology combined with machine learning to reveal the action mechanism of licochalcone intervention in liver cancer［J］. Int J Mol Sci，2023，24（21）：15935-15960. doi:10.3390/ijms242115935 doi: 10.3390/ijms242115935
4	BAO X， HU J， ZHAO Y， et al. Advances on the anti-tumor mechanisms of the carotenoid Crocin［J］. PeerJ， 2023，11：e15535. doi:10.7717/peerj.15535 doi: 10.7717/peerj.15535
5	HU X， WANG J， SHANG P， et al. Sauchinone inhibits breast cancer cell proliferation through regulating microRNA-148a-3p/HER-2 axis［J］. Thorac Cancer， 2023，14（13）：1135-1144. doi:10.1111/1759-7714.14834 doi: 10.1111/1759-7714.14834
6	KIM Y W， JANG E J， KIM C H， et al. Sauchinone exerts anticancer effects by targeting AMPK signaling in hepatocellular carcinoma cells［J］. Chem Biol Interact， 2017，261：108-117. doi:10.1016/j.cbi.2016.11.016 doi: 10.1016/j.cbi.2016.11.016
7	GAO W， LU Y X， WANG F， et al. miRNA-217 inhibits proliferation of hepatocellular carcinoma cells by regulating KLF5［J］. Eur Rev Med Pharmacol Sci， 2019，23（18）：7874-7883.
8	WANG H， HOU W， PERERA A， et al. Targeting EphA2 suppresses hepatocellular carcinoma initiation and progression by dual inhibition of JAK1/STAT3 and AKT signaling［J］. Cell Rep，2021，34（8）：108765-108805. doi:10.1016/j.celrep.2021.108765 doi: 10.1016/j.celrep.2021.108765
9	PAPACONSTANTINOU D， TSILIMIGRAS D I， PAWLIK T M. Recurrent hepatocellular carcinoma： Patterns， detection， staging and treatment［J］. J Hepatocell Carcinoma， 2022，9：947-957. doi:10.2147/jhc.s342266 doi: 10.2147/jhc.s342266
10	沈未，鲁迪，徐骁. 肝癌肝移植适应证的科学演进［J］. 器官移植，2025，16（2）：202-207.
11	CHEN C， WANG Z， DING Y， et al. Tumor microenvironment-mediated immune evasion in hepatocellular carcinoma［J］. Front Immunol， 2023，14：1133308. doi:10.3389/fimmu.2023.1133308 doi: 10.3389/fimmu.2023.1133308
12	ZHOU W， YANG F. Circular RNA circRNA-0039459 promotes the migration， invasion， and proliferation of liver cancer cells through the adsorption of miR-432［J］. Bioengineered， 2022，13（5）：11810-11821. doi:10.1080/21655979.2022.2073129 doi: 10.1080/21655979.2022.2073129
13	KIM N H， SUNG N J， SHIN S， et al. Sauchinone inhibits the proliferation， migration and invasion of breast cancer cells by suppressing Akt-CREB-MMP13 signaling pathway［J］. Biosci Rep， 2021，41（10）：BSR20211067. doi:10.1042/bsr20211067 doi: 10.1042/bsr20211067
14	LU Y， YU M， YE J， et al. Sauchinone inhibits the proliferation and immune invasion capacity of colorectal cancer cells through the suppression of PD-L1 and MMP2/MM9［J］. Anticancer Agents Med Chem，2023，23（12）：1406-1414. doi:10.2174/1871520623666230320125406 doi: 10.2174/1871520623666230320125406
15	LI S Q， FENG J， YANG M， et al. Sauchinone： A prospective therapeutic agent-mediated EIF4EBP1 down-regulation suppresses proliferation， invasion and migration of lung adenocarcinoma cells［J］. J Nat Med， 2020，74（4）：777-787. doi:10.1007/s11418-020-01435-4 doi: 10.1007/s11418-020-01435-4
16	YAO S J， MA H S， LIU G M， et al. Increased IL-1α expression is correlated with bladder cancer malignant progression［J］. Arch Med Sci， 2020，19（1）：160-170.
17	TRIPATHI A K， DESAI P P， TYAGI A， et al. Short peptides based on the conserved regions of MIEN1 protein exhibit anticancer activity by targeting the MIEN1 signaling pathway［J］. J Biol Chem， 2024，300（3）：105680-105692. doi:10.1016/j.jbc.2024.105680 doi: 10.1016/j.jbc.2024.105680
18	SUN J， HU J R， LIU C F， et al. ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis［J］. BMC Cancer， 2023，23（1）：1108. doi:10.1186/s12885-023-11612-9 doi: 10.1186/s12885-023-11612-9
19	边红艳，张舒，孟珊珊，等. 石榴皮多酚调控miR-138-5p/HIF-1α通路对结肠癌细胞恶性生物学行为的影响［J］. 实用医学杂志，2025，41（5）：676-682.
20	FORTIN B M， PFEIFFER S M， INSUA-RODRÍGUEZ J， et al. Circadian control of tumor immunosuppression affects efficacy of immune checkpoint blockade［J］. Nat Immunol， 2024，25（7）：1257-1269. doi:10.1038/s41590-024-01859-0 doi: 10.1038/s41590-024-01859-0
21	PAN Y， SHU G， FU L， et al. EHBP1L1 drives immune evasion in renal cell carcinoma through binding and stabilizing JAK1［J］. Adv Sci （Weinh），2023，10（11）：e2206792. doi:10.1002/advs.202206792 doi: 10.1002/advs.202206792
22	王芳，胡洁琼，冯彦虎，等. 乌司他丁调节CCL2-CCR2信号轴对胃癌细胞上皮-间质转化和免疫逃逸的影响分析［J］. 中国免疫学杂志，2024，40（2）：305-311.
23	HAO L， LI S， DENG J， et al. The current status and future of PD-L1 in liver cancer［J］. Front Immunol， 2023，14：1323581. doi:10.3389/fimmu.2023.1323581 doi: 10.3389/fimmu.2023.1323581
24	周智华，常静雯，严元元，等. 参芪扶正注射液增强免疫检查点抑制剂anti-PD-L1抗肿瘤作用及机制研究［J］. 中国临床药理学与治疗学，2024，29 （7）：792-799.
25	ZHAO P， SUN J， HUANG X， et al. Targeting the KLF5-EphA2 axis can restrain cancer stemness and overcome chemoresistance in basal-like breast cancer［J］. Int J Biol Sci， 2023，19（6）：1861-1874. doi:10.7150/ijbs.82567 doi: 10.7150/ijbs.82567
26	WANG Y， YU B， QU M， et al. Britannin inhibits cell proliferation， migration and glycolysis by downregulating KLF5 in lung cancer［J］. Exp Ther Med， 2024，27（3）：109-117. doi:10.3892/etm.2024.12397 doi: 10.3892/etm.2024.12397
27	XIANG Y， HUANG Y， SUN H， et al. Deregulation of miR-520d-3p promotes hepatocellular carcinoma development via lncRNA MIAT regulation and EPHA2 signaling activation［J］. Biomed Pharmacother， 2019，109：1630-1639. doi:10.1016/j.biopha.2018.11.014 doi: 10.1016/j.biopha.2018.11.014

组别	EdU阳性率（%）	OD₄₅₀值
肝癌组	28.76 ± 1.32	0.89 ± 0.07
Sch-L组	24.23 ± 1.01^?	0.76 ± 0.06^?
Sch-M组	19.92 ± 0.83^?#	0.64 ± 0.05^?#
Sch-H组	12.11 ± 0.53^?#&	0.39 ± 0.03^?#&
Sch-H + OE-NC组	12.34 ± 0.46	0.41 ± 0.03
Sch-H + OE-KLF5组	20.55 ± 0.98^@△	0.65 ± 0.04^@△
F值	315.846	95.367
P值	< 0.001	< 0.001

组别	迁移距离
肝癌组	786.65 ± 30.44
Sch-L组	626.77 ± 25.56^?
Sch-M组	392.36 ± 18.55^?#
Sch-H组	215.56 ± 9.34^?#&
Sch-H + OE-NC组	210.88 ± 8.76
Sch-H + OE-KLF5组	416.63 ± 19.81^@△
F值	756.800
P值	< 0.001

组别	细胞侵袭数
肝癌组	73.23 ± 3.21
Sch-L组	52.67 ± 2.06^?
Sch-M组	38.67 ± 1.83^?#
Sch-H组	12.43 ± 0.52^?#&
Sch-H+OE-NC组	12.55 ± 0.51
Sch-H+OE-KLF5组	40.55 ± 1.89^@△
F值	904.600
P值	< 0.001

组别	CD8⁺T细胞杀伤率
肝癌共培养组	11.26 ± 0.43
Sch-L共培养组	20.54 ± 0.82^?
Sch-M共培养组	31.18 ± 1.40^?#
Sch-H共培养组	43.69 ± 1.89^?#&
Sch-H + OE-NC共培养组	43.01 ± 1.73
Sch-H + OE-KLF5共培养组	28.63 ± 1.32^@△
F值	517.900
P值	< 0.001

组别	IFN-γ	TNF-α	IL-4
肝癌共培养组	23.31 ± 0.95	28.55 ± 1.26	34.69 ± 1.61
Sch-L共培养组	27.76 ± 1.13^?	33.68 ± 1.31^?	39.71 ± 1.80^?
Sch-M共培养组	34.55 ± 1.25^?#	39.22 ± 1.37^?#	46.68 ± 1.99^?#
Sch-H共培养组	41.88 ± 1.96^?#&	47.23 ± 2.05^?#&	55.78 ± 2.18^?#&
Sch-H+OE-NC共培养组	42.03 ± 1.89	47.63 ± 1.99	55.16 ± 2.03
Sch-H+OE-KLF5共培养组	30.66 ± 1.22^@△	36.44 ± 1.57^@△	43.78 ± 2.07^@△
F值	164.470	130.288	110.561
P值	< 0.001	< 0.001	< 0.001

Effects of sauchinone regulating KLF5⁃EphA2 pathway on the growth and immune escape of hepatocellular carcinoma cells

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 10

References 27

Related Articles 11

Recommended Articles

Metrics

Comments

组别	PCNA/GAPDH	MIEN1/GAPDH	MMP-2/GAPDH	PD-L1/GAPDH	KLF5/GAPDH	EphA2/GAPDH
肝癌组	2.56 ± 0.15	2.08 ± 0.16	1.21 ± 0.12	1.75 ± 0.15	0.91 ± 0.07	0.71 ± 0.06
Sch-L组	2.01 ± 0.18^?	1.79 ± 0.13^?	1.00 ± 0.08^?	1.34 ± 0.12^?	0.72 ± 0.06^?	0.54 ± 0.04^?
Sch-M组	1.43 ± 0.12^?#	1.32 ± 0.11^?#	0.72 ± 0.06^?#	1.01 ± 0.10^?#	0.51 ± 0.04^?#	0.38 ± 0.03^?#
Sch-H组	0.95 ± 0.07^?#&	0.83 ± 0.07^?#&	0.31 ± 0.04^?#&	0.62 ± 0.05^?#&	0.22 ± 0.02^?#&	0.11 ± 0.01^?#&
Sch-H+OE-NC组	0.97 ± 0.08	0.85 ± 0.06	0.33 ± 0.02	0.65 ± 0.05	0.23 ± 0.01	0.13 ± 0.01
Sch-H+OE-KLF5组	1.58 ± 0.13^@△	1.41 ± 0.12^@△	0.69 ± 0.05^@△	1.08 ± 0.09^@△	0.54 ± 0.04^@△	0.40 ± 0.03^@△
F值	143.000	115.800	159.200	110.100	216.400	270.300
P值	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

[1]	Xinfa LIU,Yin YUAN,Chunmei LUAN,Lin. JIANG. Analysis of the predictive value of bispectral index dynamic changes for hypotension after laparoscopic surgery in elderly patients with primary liver cancer [J]. The Journal of Practical Medicine, 2025, 41(10): 1460-1465.
[2]	Hong YANG,Jun DING,Peng GUO,Zhiqing. ZHANG. The predictive value of combined detection of cell cycle molecule CDKN3 and serum CHI3L1， miR⁃106b in liver cancer metastasis [J]. The Journal of Practical Medicine, 2025, 41(10): 1473-1479.
[3]	Yanping TANG,Kezhi LI,Zhengmin CAI,Hao TAO,Jiaying TANG,Xueyu LI,Yanjuan LI,Ji CAO. Study on the mechanism of fatty acid binding protein 5 binding Vimentin protein in the Hepatocellular Carcinoma cells [J]. The Journal of Practical Medicine, 2024, 40(6): 756-761.
[4]	Huiling YE,Renming. ZHONG. Progress in assessing the treatment accuracy of liver stereotactic body radiotherapy through post-therapeutic magnetic resonance imaging morphologic alterations [J]. The Journal of Practical Medicine, 2024, 40(22): 3119-3123.
[5]	Xinyi ZHANG,Guoliang ZHANG,Guanglin. ZHANG. Clinical value of quantitative detection of serum HBsAg and HBeAg to evaluate the viral response of entecavir in patients with hepatitis B cirrhosis complicated with primary liver cancer [J]. The Journal of Practical Medicine, 2023, 39(17): 2241-2247.
[6]	YANG Kailun, LI Zhi, REN Qimeng, LIU Zhao, MAO Xinyu, ZHU Xiaoli, NI Caifang. . The efficacy and safety of TACE with different chemotherapy regimens in the treatment of primary liver cancer [J]. The Journal of Practical Medicine, 2022, 38(5): 594-599.
[7]	ZHANG Xiaokun, SITU Ruiru.. Clinical value of combined detection of alpha⁃fetoprotein，alpha⁃fetoprotein alloplasm⁃3 ratio and Golgi so⁃ ma protein 73 in primary liver cancer [J]. The Journal of Practical Medicine, 2021, 37(8): 1068-1071.
[8]	WANG Shaoshuang, ZHOU Yannan, YANG Tingting, YAN Jun, DONG Maijuan, WANG Qiang.. Effect of general anesthesia combined with regional block on analgesia and postoperative rehabilitation in liver cancer patients undergoing laparotomy Effect of general anesthesia combined with regional block on analgesia and postoperative rehabilitation in liver cancer patients undergoing laparotomy [J]. The Journal of Practical Medicine, 2021, 37(15): 1939-1943.
[9]	LI Xilong, LI Changsheng, WANG Maoai, YANG Chunyao, ZHANG Zhen, MIAO Changhong, LU Xihua. . Effect of dexmedetomidine on inflammatory response and perioperative neurocognitive disorder in elderly patients undergoing laparoscopic radiofrequency ablation for liver cancer [J]. The Journal of Practical Medicine, 2021, 37(15): 1999-2002.
[10]	YANGPeiyu, DU Wei, LI Zhengliang, ZHOU Zhou, XIONG Wencui. . Effect of WY⁃14643 on oxidative stress and apoptosis of liver tissues adjacent to cancer after TAE [J]. The Journal of Practical Medicine, 2021, 37(13): 1660-1665.
[11]	GAO Wenhui, WU Jinjun, JIAN Xiaoshun, JIA Xiaoting, LI Jing. . Preparation of sorafenib loaded human serum albumin nanoparticles and evaluation of their anti ⁃hepatic tumor activity in vitro [J]. The Journal of Practical Medicine, 2021, 37(11): 1392-1396.