Epicatechin attenuates the stemness of liver cancer stem cells and tumorigenesis through DNA methylation-mediated inactivation of GINS1/HRAS

Abstract Background The limited therapeutic options for hepatocellular carcinoma (HCC) are primarily attributed to the presence of liver cancer stem cells (LCSCs). Epicatechin (EC) is known to exert therapeutic effects on cancer. Go-ichi-ni-san complex subunit 1 (GINS1) is an oncogene associated wit...

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Main Authors: Xingbao Fang, Yan Cai, Xiongbing Peng, Zhaojun Li, Meifang Huang, Yuehong Li, Peiwan Liu
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Journal of Translational Medicine
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Online Access:https://doi.org/10.1186/s12967-025-06790-y
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author Xingbao Fang
Yan Cai
Xiongbing Peng
Zhaojun Li
Meifang Huang
Yuehong Li
Peiwan Liu
author_facet Xingbao Fang
Yan Cai
Xiongbing Peng
Zhaojun Li
Meifang Huang
Yuehong Li
Peiwan Liu
author_sort Xingbao Fang
collection DOAJ
description Abstract Background The limited therapeutic options for hepatocellular carcinoma (HCC) are primarily attributed to the presence of liver cancer stem cells (LCSCs). Epicatechin (EC) is known to exert therapeutic effects on cancer. Go-ichi-ni-san complex subunit 1 (GINS1) is an oncogene associated with HCC and represents a potential target for cancer therapy. In this study, we investigated the molecular mechanisms of EC by which modulates DNA methylation and inhibits GINS1, thereby affecting LCSCs and HCC malignancy. Methods Cancer Genome Atlas (TCGA) database was employed to analyze the differences in GINS1 and HRAS expression levels between HCC tissues and normal tissues. The effect of EC on GINS1 and HRAS expression was measured by qPCR and western blotting. Sphere formation assay, CCK-8 assays and transwell assays were used to evaluate the malignant behaviors of HCC cells in vitro. Stemness markers (Nanog, OCT4 and SOX2) were analyzed by western blotting. Methylation-specific PCR was conducted to assess DNA methylation level on GINS1 promoter. The correlation between GINS1 and HRAS in HCC was evaluated using spearman’s rank correlation analysis and confirmed through co-immunoprecipitation and GST pulldown assay. Additionally, xenograft tumor models based on HCC cells were conducted to investigate the anti-tumor effects of EC in vivo. Results Elevated expression of GINS1 was observed in HCC cells and LCSCs. Silencing GINS1 repressed LCSC phenotype, diminished the malignant behaviors of HCC cells and inhibited tumorigenesis. We further found EC attenuated LCSC phenotype and tumorigenesis by downregulating GINS1 expression. Mechanistically, administration of EC enhanced DNA methylation on the GINS1 promoter, leading to a subsequent reduction in GINS1 levels. Additionally, we found that GINS1 could interact with HRAS and activate HRAS, thereby inducing LCSC phenotype and promoting tumorigenesis. Overexpression of HRAS partially counteracted the inhibitory effects on LCSC phenotype induction and tumorigenesis mediated by GINS1 silencing or EC administration. Conclusion Taken together, our findings suggest an important role of the EC/DNA methylation/GINS1/HRAS pathway in regulating LCSCs properties, suggesting potential therapeutic targets for HCC.
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spelling doaj-art-ca4f24b11cfe4158804bedb3f08ee3b02025-08-20T03:46:12ZengBMCJournal of Translational Medicine1479-58762025-07-0123111510.1186/s12967-025-06790-yEpicatechin attenuates the stemness of liver cancer stem cells and tumorigenesis through DNA methylation-mediated inactivation of GINS1/HRASXingbao Fang0Yan Cai1Xiongbing Peng2Zhaojun Li3Meifang Huang4Yuehong Li5Peiwan Liu6Hepatobiliary Pancreatic Surgery, Kunming Medical University Affiliated Qujing Hospital The First People’s Hospital of QujingPathology Teaching and Research Office, Qujing Medical CollegeHepatobiliary Pancreatic Surgery, Kunming Medical University Affiliated Qujing Hospital The First People’s Hospital of QujingHepatobiliary Pancreatic Surgery, Kunming Medical University Affiliated Qujing Hospital The First People’s Hospital of QujingDepartment of Oncology, Kunming Medical University Affiliated Qujing Hospital The First People’s Hospital of QujingHepatobiliary Pancreatic Surgery, Kunming Medical University Affiliated Qujing Hospital The First People’s Hospital of QujingHepatobiliary Pancreatic Surgery, Kunming Medical University Affiliated Qujing Hospital The First People’s Hospital of QujingAbstract Background The limited therapeutic options for hepatocellular carcinoma (HCC) are primarily attributed to the presence of liver cancer stem cells (LCSCs). Epicatechin (EC) is known to exert therapeutic effects on cancer. Go-ichi-ni-san complex subunit 1 (GINS1) is an oncogene associated with HCC and represents a potential target for cancer therapy. In this study, we investigated the molecular mechanisms of EC by which modulates DNA methylation and inhibits GINS1, thereby affecting LCSCs and HCC malignancy. Methods Cancer Genome Atlas (TCGA) database was employed to analyze the differences in GINS1 and HRAS expression levels between HCC tissues and normal tissues. The effect of EC on GINS1 and HRAS expression was measured by qPCR and western blotting. Sphere formation assay, CCK-8 assays and transwell assays were used to evaluate the malignant behaviors of HCC cells in vitro. Stemness markers (Nanog, OCT4 and SOX2) were analyzed by western blotting. Methylation-specific PCR was conducted to assess DNA methylation level on GINS1 promoter. The correlation between GINS1 and HRAS in HCC was evaluated using spearman’s rank correlation analysis and confirmed through co-immunoprecipitation and GST pulldown assay. Additionally, xenograft tumor models based on HCC cells were conducted to investigate the anti-tumor effects of EC in vivo. Results Elevated expression of GINS1 was observed in HCC cells and LCSCs. Silencing GINS1 repressed LCSC phenotype, diminished the malignant behaviors of HCC cells and inhibited tumorigenesis. We further found EC attenuated LCSC phenotype and tumorigenesis by downregulating GINS1 expression. Mechanistically, administration of EC enhanced DNA methylation on the GINS1 promoter, leading to a subsequent reduction in GINS1 levels. Additionally, we found that GINS1 could interact with HRAS and activate HRAS, thereby inducing LCSC phenotype and promoting tumorigenesis. Overexpression of HRAS partially counteracted the inhibitory effects on LCSC phenotype induction and tumorigenesis mediated by GINS1 silencing or EC administration. Conclusion Taken together, our findings suggest an important role of the EC/DNA methylation/GINS1/HRAS pathway in regulating LCSCs properties, suggesting potential therapeutic targets for HCC.https://doi.org/10.1186/s12967-025-06790-yHepatocellular carcinomaCancer stem cellEpicatechinDNA methylationGINS1HRAS
spellingShingle Xingbao Fang
Yan Cai
Xiongbing Peng
Zhaojun Li
Meifang Huang
Yuehong Li
Peiwan Liu
Epicatechin attenuates the stemness of liver cancer stem cells and tumorigenesis through DNA methylation-mediated inactivation of GINS1/HRAS
Journal of Translational Medicine
Hepatocellular carcinoma
Cancer stem cell
Epicatechin
DNA methylation
GINS1
HRAS
title Epicatechin attenuates the stemness of liver cancer stem cells and tumorigenesis through DNA methylation-mediated inactivation of GINS1/HRAS
title_full Epicatechin attenuates the stemness of liver cancer stem cells and tumorigenesis through DNA methylation-mediated inactivation of GINS1/HRAS
title_fullStr Epicatechin attenuates the stemness of liver cancer stem cells and tumorigenesis through DNA methylation-mediated inactivation of GINS1/HRAS
title_full_unstemmed Epicatechin attenuates the stemness of liver cancer stem cells and tumorigenesis through DNA methylation-mediated inactivation of GINS1/HRAS
title_short Epicatechin attenuates the stemness of liver cancer stem cells and tumorigenesis through DNA methylation-mediated inactivation of GINS1/HRAS
title_sort epicatechin attenuates the stemness of liver cancer stem cells and tumorigenesis through dna methylation mediated inactivation of gins1 hras
topic Hepatocellular carcinoma
Cancer stem cell
Epicatechin
DNA methylation
GINS1
HRAS
url https://doi.org/10.1186/s12967-025-06790-y
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