Plant Flavonoids from Coreopsis tinctoria Show Multi-Target Power Against Liver Cancer
Network pharmacology and lab experiments reveal CTFs suppress hepatocellular carcinoma via apoptosis, ROS induction, and cell cycle arrest.
Summary
Researchers combined computational network pharmacology with laboratory experiments to investigate how total flavonoids from Coreopsis tinctoria (CTFs) fight hepatocellular carcinoma (HCC). They identified 27 bioactive compounds and 32 HCC-relevant targets, with core targets including Caspase-3, p53, Bcl-2, Bax, and MAPK1. Key flavonoids—quercetin, EGCG, fisetin, acacetin, luteolin, and kaempferol—showed strong molecular docking scores against these targets. In HepG2 liver cancer cells, CTFs inhibited proliferation, triggered reactive oxygen species production, induced G2/M and S-phase cell cycle arrest, and promoted apoptosis. Normal liver cells (LO2) were unaffected or stimulated. A zebrafish xenograft model confirmed significant tumor suppression in vivo. These findings position CTFs as a promising multi-target botanical therapeutic candidate for HCC.
Detailed Summary
Hepatocellular carcinoma remains one of the deadliest cancers globally, with limited effective treatments and high recurrence rates. Natural plant compounds, particularly flavonoids, have attracted growing interest as multi-target anticancer agents with potentially favorable safety profiles. This study systematically investigated the anti-HCC potential of Coreopsis tinctoria Total Flavonoids (CTFs), a botanical extract used in traditional Uyghur medicine, using an integrated network pharmacology and experimental validation approach.
Using databases including TCMSP, GeneCards, OMIM, and DrugBank, researchers identified 27 bioactive CTF components and 318 putative molecular targets, of which 32 overlapped with HCC-associated genes. Protein-protein interaction (PPI) network analysis and Cytoscape visualization highlighted core targets: Caspase-3, p53, MAPK1, Bcl-2, and Bax. The most pharmacologically active compounds were quercetin, (-)-epigallocatechin (EGCG), fisetin, acacetin, luteolin, and kaempferol. Molecular docking via Autodock Vina confirmed strong binding affinities between these flavonoids and the core targets. KEGG and GO enrichment analyses pointed to apoptosis, cell cycle regulation, and oxidative stress pathways as primary mechanisms of action.
Bioinformatic analysis of HCC versus normal tissue showed that pro-apoptotic genes Bax, Caspase-3, and p53 were upregulated in tumor tissue, and Kaplan-Meier survival analysis revealed that low expression of Bax and Caspase-3 correlated with significantly worse patient outcomes—validating these as clinically meaningful targets.
In vitro experiments using HepG2 hepatocellular carcinoma cells demonstrated that CTFs dose-dependently inhibited proliferation (CCK-8 assay), reduced colony formation, induced reactive oxygen species (ROS) production, caused G2/M and S-phase cell cycle arrest, and promoted apoptosis (flow cytometry). Importantly, CTFs promoted rather than inhibited growth in normal LO2 hepatocytes, suggesting selective cytotoxicity. qPCR confirmed upregulation of Bax, p53, and Caspase-3, and downregulation of Bcl-2, with an increased Bax/Bcl-2 ratio consistent with pro-apoptotic signaling. In vivo validation using a zebrafish xenograft model showed significant tumor growth suppression at non-toxic concentrations, confirmed by fluorescence imaging and HE staining.
These findings collectively support CTFs as a multi-target botanical agent capable of engaging apoptotic, oxidative stress, and cell cycle pathways simultaneously. The selective action on cancer versus normal liver cells is particularly noteworthy. While the study provides compelling preclinical evidence, mammalian animal models and eventual clinical trials will be necessary to confirm translational relevance. The work also opens avenues for isolating and optimizing individual flavonoid components for maximum therapeutic efficacy.
Key Findings
- CTFs identified 27 bioactive compounds targeting 32 HCC-relevant genes including Caspase-3, p53, Bcl-2, Bax, and MAPK1.
- Molecular docking confirmed strong binding of quercetin, EGCG, fisetin, luteolin, and kaempferol to core HCC targets.
- CTFs selectively inhibited HepG2 cancer cell proliferation while promoting normal LO2 hepatocyte growth.
- CTFs induced ROS production, G2/M and S-phase arrest, and apoptosis, with upregulated Bax/Caspase-3 and suppressed Bcl-2.
- Zebrafish xenograft model confirmed significant in vivo tumor suppression at non-toxic CTF concentrations.
Methodology
The study used network pharmacology (TCMSP, GeneCards, STRING, Cytoscape, DAVID) to map CTF-HCC target interactions, followed by molecular docking validation. In vitro experiments (CCK-8, flow cytometry, qPCR, colony formation) in HepG2 and LO2 cells and a zebrafish xenograft in vivo model provided experimental confirmation.
Study Limitations
The study relies primarily on cell lines and zebrafish models; mammalian (rodent) in vivo studies and pharmacokinetic data are absent. Network pharmacology predictions require further wet-lab validation for each identified target. Clinical translation remains distant without toxicology and efficacy data in higher organisms.
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