Spectrum-effect relationship between HPLC fingerprints and antitumor activities of Inonotus hispidus

Inonotus hispidus (I. hispidus), a traditional edible and medicinal fungus, has been widely used in the fields of medicine and health care. Modern pharmacological studies have confirmed that this fungus exhibits various biological activities, including antitumor, antioxidant, and hypoglycemic effects. However, the underlying mechanisms linking its antitumor activity to specific chemical constituents have not yet been fully elucidated. In this study, HPLC fingerprints of 11 batches of I. hispidus were established. The inhibitory effects of these samples on 4 T1 and HeLa cells were evaluated using the MTT assay. The correlation between the HPLC fingerprints and antitumor activity was analyzed using grey relational analysis and partial least squares regression. The results indicated that the antitumor activity of I. hispidus was due to the synergistic effects of multiple components. Of these, peaks 3, 5 (hispidin), 6, and 7 were identified as key contributors. Notably, these key components were predominantly enriched in the ethyl acetate fraction. The accuracy of the correlation results was validated by performing in vitro inhibition of 4 T1 cell viability using different extraction solvents. This study provides scientific evidence for the antitumor potential of I. hispidus and helps clarify the spectrum-effect relationship of its active components.