Related Experiment Video
Updated: Nov 9, 2025
Discovery of Driver Genes in Colorectal HT29-derived Cancer Stem-Like Tumorspheres
Published on: July 22, 2020
Transcriptome analysis of signaling pathways targeted by Ellagic acid in hepatocellular carcinoma cells
Shuang Qiu1, Chen Zhong2, Bo Zhao1
1Key Laboratory of Saline-alkali Vegetation Ecology Restoration, Ministry of Education, College of Life Science, Northeast Forestry University, Harbin 150040, China.
Background:
Ellagic acid (EA) possesses prominent inhibitory activities against various cancers, including hepatocellular carcinoma (HCC). Our recent study demonstrated EA's activities in reducing HCC cell proliferation and tumor formation. However, the mechanisms of EA to exert its anticancer activities and its primary targets in cancer cells have not been systematically explored.
Methods:
Cell proliferation assay and flow cytometric analysis were used to examine the effects of EA treatment on viability and apoptosis, respectively, of HepG2 cells. RNA-seq studies and associated pathway analyses by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were employed to determine EA's primary targets. Differentially expressed genes (DEG) in EA-treated HepG2 cells were verified by RT-qPCR and Western blot. Integrative analyses of the RNA-seq dataset with a TCGA dataset derived from HCC patients were conducted to verify EA-targeted genes and signaling pathways. Interaction network analysis of the DEGs, shRNA-mediated knockdown, cell viability assay, and colony formation assay were used to validate EA's primary targets.
Results:
EA reduced cell viability, caused DNA damage, and induced cell cycle arrest at G1 phase of HepG2 cells. We identified 5765 DEGs encoding proteins with over 2.0-fold changes in EA-treated HepG2 cells by DESeq2. These DEGs showed significant enrichment in the pathways regulating DNA replication and cell cycle progression. As primary targets, p21 was significantly upregulated, while MCM2-7 were uniformly downregulated in response to EA treatment. Consistently, p21 knockdown desensitized liver cells to EA in cell viability and colony formation assays.
Conclusion:
EA induced G1 phase arrest and promoted apoptosis of HCC cells through activating the p21 gene and downregulating the MCM2-7 genes, respectively.
General Significance:
The discoveries in this study provide helpful insights into developing novel strategies in the therapeutic treatment of HCC patients.
Insights
Ellagic acid (EA) inhibits hepatocellular carcinoma (HCC) by inducing cell cycle arrest via p21 activation and promoting apoptosis through MCM2-7 downregulation. This research clarifies EA's anticancer mechanisms for potential HCC therapies.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Ellagic acid (EA) demonstrates significant anticancer properties, particularly against hepatocellular carcinoma (HCC).
- Previous research confirmed EA's ability to reduce HCC cell proliferation and tumor formation.
- The precise molecular mechanisms and primary cellular targets of EA's anticancer effects remained largely unexplored.
Purpose of the Study:
- To systematically investigate the molecular mechanisms underlying EA's anticancer activities in HCC.
- To identify the primary molecular targets of EA within cancer cells.
- To validate the role of identified targets in EA-mediated anti-HCC effects.
Main Methods:
- Utilized HepG2 cell line for in vitro studies, employing cell proliferation and flow cytometry assays.
- Conducted RNA-sequencing (RNA-seq) for global gene expression analysis, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses.
- Validated differentially expressed genes (DEGs) using RT-qPCR and Western blot, and integrated findings with TCGA HCC patient data.
- Employed shRNA-mediated knockdown and colony formation assays to confirm EA's primary targets.
Main Results:
- EA treatment led to reduced HepG2 cell viability, DNA damage, and G1 phase cell cycle arrest.
- RNA-seq identified 5765 differentially expressed genes (DEGs) enriched in DNA replication and cell cycle pathways.
- Key findings revealed significant upregulation of p21 and uniform downregulation of MCM2-7 genes in response to EA.
- Knockdown of p21 diminished EA's effects on cell viability and colony formation, confirming its role.
Conclusions:
- EA effectively induces G1 phase cell cycle arrest and promotes apoptosis in HCC cells.
- These effects are mediated by the activation of the p21 gene and the downregulation of MCM2-7 genes.
- The study provides crucial insights into EA's therapeutic potential for HCC treatment.
