Published online Jan 28, 2026. doi: 10.3748/wjg.v32.i4.114420
Revised: October 28, 2025
Accepted: December 8, 2025
Published online: January 28, 2026
Processing time: 126 Days and 5.4 Hours
Liver carcinoma, as a major global health concern due to its high incidence and mortality rates. Despite advancements in diagnostic and treatment methodologies, outcomes for hepatocellular carcinoma remain unsatisfactory. In response to these limitations, patients increasingly turn to alternative therapies such as traditional Chinese medicine, which has demonstrated potential in enhancing quality of life and prolonging survival in combination with conventional treat
To investigate the synergistic anti-hepatoma effect of TP and quercetin and elucidate the underlying molecular mechanism involving the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) and mam
The study utilized 5-week-old female BALB/c-nu mice for establishing a liver cancer subcutaneous transplant tumor model. TP and quercetin were administered intraperitoneally over 21 days to evaluate the effectiveness of the combination, with monitoring of tumor growth. IncuCyte Zoom and CompuSyn software were employed to analyze drug effects for different dose combination on cell proliferation and synergy. Various assays such as CCK-8 cell proliferation analysis, plate cell clone formation, cell scratch experiments, Transwell migration and invasion assays, Annexin V-FITC flow cytometry, and western blotting using specific antibodies were employed to assess cell apoptosis, migration, invasion. Then transcriptome analysis was used RNA sequencing to find the potential synergistic mechanisms and proved by western blotting.
In vivo, the combination therapy significantly slowed down tumor growth compared to the control group, quercetin alone group, and TP alone group. The tumor inhibition rates were 28.91% (quercetin), 28.8% (TP), and 59.3% (combination therapy), respectively. The determination of IncuCyte Zoom and CCK-8 confirmed that there is a concentration gradient and time gradient effect on tumor inhibition, with the synergistic effect of 25 nmol/L TP and 100 μmol/L quercetin being the best. Platelet cell clone formation and cell wound scratch assay showed that the combination group had better inhibitory effects. Transwell analysis showed a decrease in migration and invasion in the combination therapy group. Flow cytometry showed that over time, cell apoptosis increased after combination therapy. Transcriptome analysis emphasizes unique pathways influenced by the combination (JAK-STAT and mTOR signaling pathways) and has been validated at the protein level.
Compared with a single drug, the specific metering combination of TP and quercetin has enhanced anti-tumor effects, mediated by inhibition of cell proliferation, inducing cell apoptosis and inhibiting migration/invasion. This synergistic effect is closely related to the simultaneous inhibition of signaling pathways JAK-STAT and mTOR concurrently.
Core Tip: This study demonstrates that a specific combination of triptolide (25 nmol/L) and quercetin (100 μmol/L) exerts a synergistic antitumor effect against hepatocellular carcinoma both in vitro and in vivo. The combination significantly inhibits tumor cell proliferation, induces apoptosis, and suppresses migration and invasion more effectively than either drug alone. Mechanistically, this synergy is achieved through the simultaneous inhibition of the Janus kinase-signal transducer and activator of transcription and mammalian target of rapamycin signaling pathways, as confirmed by transcriptomic analysis, molecular docking, and western blot validation. Importantly, the combination reduces the required dose of triptolide, thereby mitigating its toxic side effects while maintaining potent anticancer efficacy. This research highlights a promising therapeutic strategy for hepatocellular carcinoma that leverages the complementary actions of natural compounds to enhance treatment outcomes.