Screening Prevention and Treatment of Liver Cancer Quality Markers (Q-Markers) from Ganoderma lucidum Spore (GLS) Based on Spectrum–Effect Relationship

Liver cancer remains a major global health challenge with limited treatment options, and oxidative stress is increasingly recognized as a key driver of liver cancer development. Ganoderma lucidum spores (GLS) have demonstrated anti-liver cancer efficacy in clinical settings; however, their quality is highly variable due to undefined quality markers (Q-markers). This study established UPLC-Q-TOF-MS/MS and FT-IR fingerprint profiles for GLS from different origins and correlated them with pharmacodynamic parameters in an H22 tumor-bearing mouse model through spectrum–effect relationship analysis. Five triterpenoids, namely ganoderic acids A, B, G, and ganoderenic acids E and D, were identified as efficacy-correlated components. All five compounds inhibited the proliferation of HepG-2 and H22 cells, and component knockout assays demonstrated that the removal of any single component significantly impaired the combined anticancer efficacy. The Q-marker mixture dose-dependently reduced intracellular ROS accumulation in HepG-2 cells, and chemical antioxidant assays (DPPH, ABTS, FRAP) demonstrated that the Q-marker mixture exhibited significantly stronger free radical scavenging and iron-reducing capabilities compared to the crude GLS extract. The network pharmacology analysis revealed that the PI3K/Akt signaling pathway was of great importance and played a key role in mediating oxidative stress. Subsequent verification via Western blot showed that the mixture of selected Q-markers downregulated the phosphorylation levels of PI3K and Akt in a dose-dependent manner. In summary, these results confirm that these five triterpenoids are candidate Q-markers responsible for the anticancer effect of GLS, which is mediated, at least in part, by their antioxidant activity and by the modulation of the PI3K/Akt signaling pathway, and provide a scientific basis for quality control related to the therapeutic efficacy of GLS.