Predicted mechanisms of Huaier in hepatocellular carcinoma therapy via IL-2/PI3K-Akt immunomodulation: An integrated...

Huaier (Trametes robiniophila Murr.; syn. Vanderbylia robiniophila (Murrill) B.K. Cui & Y.C. Dai), a medicinal fungus, has been traditionally used in China for adjuvant treatment of hepatocellular carcinoma (HCC). While promising, comprehensive high-quality evidence for its efficacy and a systematic understanding of its mechanisms remain limited.

This study aims to provide preliminary evidence for the clinical efficacy of Huaier through a meta-analysis, and explore its potential mechanisms of action by integrating network pharmacology, bioinformatics, molecular docking, molecular dynamics simulation, and cell experiments. By bridging clinical outcomes with molecular targets and pathways, we seek to propose a working hypothesis for the pharmacological basis of Huaier in HCC, which requires further validation.

A meta-analysis of Huaier combined with conventional antitumor therapy for HCC was constructed. Meanwhile, network pharmacology was employed to screen key targets and signaling pathways, subsequently validating them through TCGA/GTEx bioinformatics analysis, molecular docking, molecular dynamics simulation, and in vitro experiments in Jurkat cells.

The meta-analysis encompassed 27 studies comprising 2880 patients (1632 experimental; 1248 control). These findings indicated that combination therapy with Huaier improves the objective response rate (RR = 1.40, 95% CI = 1.29-1.52, P < 0.00001), disease control rate (RR = 1.15, 95% CI = 1.10-1.20, P < 0.00001), 6-month survival rate (RR = 1.20, 95% CI = 1.10-1.32, P < 0.0001), and 1-year survival rate (RR = 1.33, 95% CI = 1.20-1.48, P < 0.00001). Moreover, this therapy showed a trend toward reduced recurrence (RR = 0.74, 95% CI = 0.65-0.85, P < 0.0001) and augmented the immune function. Network pharmacology showed that Huaier may potentially exert its antitumor effects via core genes including IL-2, IL-6, TNF and VEGFA, with key pathways encompassing the PI3K-Akt, VEGF and TCR signaling pathways. Bioinformatics analysis demonstrated marked expression differences of these genes between HCC and normal tissues. Molecular docking analysis illustrated that Huaier constituents exhibited strong binding affinity to PIK3CA and AKT1. Subsequent molecular dynamics simulations further validated the stable binding of representative Huaier constituents to PIK3CA and AKT1, with MM-GBSA binding free energies ranging from -65.00 to -22.12 kcal/mol. Cellular assays indicated that Huaier upregulates IL-2 mRNA expression at the transcriptional level in Jurkat cells-an effect abrogated by the PI3K inhibitor LY294002-suggesting a potential involvement of targets associated with the PI3K-Akt signaling cascade.

The findings suggest that combination therapy with Huaier may enhance the efficacy of conventional antitumor treatments against HCC, providing preliminary evidence for its clinical application. The immunomodulatory mechanism may involve the regulation of IL-2 expression through the PI3K-Akt pathway, as supported by computational predictions and preliminary in vitro validation at the transcriptional level. However, these mechanistic conclusions remain preliminary and require further confirmation through protein-level assays and more physiologically relevant in vivo models.