Exploring the Molecular Mechanism of Theaflavin in Inhibiting Hepatocellular Carcinoma Through Reverse Virtual Screening and Biological Experimental Validation
- 1Key Laboratory of Infection and Immunity, Health Commission of Jiangxi Province, School of Basic Medicine, Nanchang Medical College, Nanchang 330052, China
Abstract
Hepatocellular carcinoma (HCC) ranks among the leading causes of cancer death, underscoring the pressing need for therapeutic agents with better efficacy and lower toxicity. Theaflavin (TF) is a polyphenol abundant in black tea that possesses broad-spectrum anti-tumor activity. However, the direct targets and molecular mechanisms of TF in HCC have not been completely defined. Here, reverse virtual screening was employed to predict candidate targets of TF, which were then validated by molecular docking, molecular dynamics (MD) simulation, drug affinity responsive target stability (DARTS), thermal shift assay (TSA) and endogenous fluorescence spectroscopy. The fluorescence titration results showed that TF could directly bind to CDK2, JNK1 and JNK2, with binding free energies of −37.08, −38.77 and −37.27 kJ/mol, and binding constants Ka of 3.16 × 106, 6.25 × 106 and 3.42 × 106 L⋅mol−1, respectively. TF inhibited the viability of HepG2, SK-Hep-1 and Huh-7 cells in a dose-dependent manner, enhanced reactive oxygen species (ROS) accumulation, reduced mitochondrial membrane potential, elevated the Bax/Bcl-2 ratio and activated caspase-9/3, thereby inducing apoptosis through the mitochondrial pathway. Moreover, TF induced G0/G1-phase arrest and curtailed cell migration and invasion, although the latter effects may be partly attributable to reduced cell viability. In nude mice bearing subcutaneous xenografts, TF markedly restrained tumor growth and promoted apoptosis. In summary, TF, a multi-target compound potentially interacting with CDK2 and JNK1/2, was associated with alterations in cell cycle- and MAPK-related signaling and may serve as a potential lead compound for HCC treatment.
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BibTeX
@article{lv2026exploring,
title = {{Exploring the Molecular Mechanism of Theaflavin in Inhibiting Hepatocellular Carcinoma Through Reverse Virtual Screening and Biological Experimental Validation}},
author = {Shaoli Lv and Bowei Zhang and Wenli Jiang and Jingyi Liu and Sitong Jie and Zonghan Li and Xiaoman Yan and Huaqing He and Xinling Liao and Wang Wang},
journal = {Biomolecules},
year = {2026},
month = nov,
doi = {10.3390/biom16111618},
url = {https://www.mdpi.com/2218-273X/16/11/1618},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Exploring the Molecular Mechanism of Theaflavin in Inhibiting Hepatocellular Carcinoma Through Reverse Virtual Screening and Biological Experimental Validation
AU - Shaoli Lv
AU - Bowei Zhang
AU - Wenli Jiang
AU - Jingyi Liu
AU - Sitong Jie
AU - Zonghan Li
AU - Xiaoman Yan
AU - Huaqing He
AU - Xinling Liao
AU - Wang Wang
PY - 2026
DA - 2026/11/06
JO - Biomolecules
DO - 10.3390/biom16111618
UR - https://www.mdpi.com/2218-273X/16/11/1618
AB - Hepatocellular carcinoma (HCC) ranks among the leading causes of cancer death, underscoring the pressing need for therapeutic agents with better efficacy and lower toxicity. Theaflavin (TF) is a polyphenol abundant in black tea that possesses broad-spectrum anti-tumor activity. However, the direct targets and molecular mechanisms of TF in HCC have not been completely defined. Here, reverse virtual screening was employed to predict candidate targets of TF, which were then validated by molecular docking, molecular dynamics (MD) simulation, drug affinity responsive target stability (DARTS), thermal shift assay (TSA) and endogenous fluorescence spectroscopy. The fluorescence titration results showed that TF could directly bind to CDK2, JNK1 and JNK2, with binding free energies of −37.08, −38.77 and −37.27 kJ/mol, and binding constants Ka of 3.16 × 106, 6.25 × 106 and 3.42 × 106 L⋅mol−1, respectively. TF inhibited the viability of HepG2, SK-Hep-1 and Huh-7 cells in a dose-dependent manner, enhanced reactive oxygen species (ROS) accumulation, reduced mitochondrial membrane potential, elevated the Bax/Bcl-2 ratio and activated caspase-9/3, thereby inducing apoptosis through the mitochondrial pathway. Moreover, TF induced G0/G1-phase arrest and curtailed cell migration and invasion, although the latter effects may be partly attributable to reduced cell viability. In nude mice bearing subcutaneous xenografts, TF markedly restrained tumor growth and promoted apoptosis. In summary, TF, a multi-target compound potentially interacting with CDK2 and JNK1/2, was associated with alterations in cell cycle- and MAPK-related signaling and may serve as a potential lead compound for HCC treatment.
SN - 2218-273X
ER -