A Multi-Level Study on the Anti-Lung Cancer Mechanism of Peiminine, a Key Component of Fritillaria ussuriensis Maxim.: Integrating Quality Analysis, Network Pharmacology, Bioinformatics Analysis, and Experimental Validation

Abstract

Globally, lung cancer is the primary cause of deaths associated with cancer; however, current therapies are costly and toxic, highlighting the need for novel treatments. Peiminine (Verticinone), a key bioactive compound derived from Fritillaria ussuriensis Maxim., has demonstrated diverse biological activities. However, the precise pharmacological mechanisms underlying its anti-lung cancer effects remain unclear. The objective of this study was to quantify the content of peiminine in Fritillaria ussuriensis Maxim. from different geographical regions using UHPLC-MS/MS and to elucidate the anti-lung cancer mechanisms of peiminine through network pharmacology, bioinformatics, and in vitro experiments. The content of peiminine in Fritillaria ussuriensis Maxim. from various regions was determined using UHPLC-MS/MS. Potential target genes associated with peiminine and lung cancer were systematically screened from multiple databases. To identify core genes, we set up a PPI (protein-protein interaction) network, followed by in-depth analyses of their corresponding target proteins. Survival analysis, molecular docking, and dynamics simulations were used to explore potential anti-cancer mechanisms. In vitro experiments on human H1299 NSCLC cells assessed peiminine's anti-tumor activity and measured key gene transcription levels. UHPLC-MS/MS analysis revealed that Fritillaria ussuriensis Maxim. from Mudanjiang (Heilongjiang Province) exhibited the highest peiminine content. Network pharmacological analysis identified PIK3CG, SRC, JAK3, AKT2, and PRKCA as key potential targets of peiminine in lung cancer treatment. Molecular docking results demonstrated strong binding affinities between peiminine and PIK3CG, SRC, and JAK3; these results were further confirmed using molecular dynamics simulations. Survival analysis indicated that a high AKT2 and PRKCA expression correlated with bad prognosis in lung cancer patients. In vitro, peiminine inhibited H1299 cell viability and regulated genes involved in the PI3K-Akt pathway (PI3K, AKT, and PTEN) and apoptosis (Bcl-2, Bax), suggesting that it may induce its effects via PI3K-Akt pathway inhibition. Peiminine from Fritillaria ussuriensis Maxim. exhibits significant anti-lung cancer potential by targeting key genes such as PIK3CG, SRC, and JAK3, as well as by modulating the PI3K-Akt signaling pathway and apoptosis-related genes. These results lay a foundation for further investigations into peiminine as a potentially effective therapeutic option for treating lung cancer. Additionally, the identified targets (PIK3CG, SRC, JAK3, AKT2, and PRKCA) may function as possible biomarkers for predicting lung cancer prognosis and guiding personalized therapy.

Keywords: PI3K–Akt signaling pathway; lung cancer; molecular docking; network pharmacology; peiminine.

Figure 1

(A) Product ion mass sepctra and structure of peiminine. (B) Representative chromatograms of peiminine reference solution. (C) Representative chromatogram of test solution of samples. (D) Heat map of peiminine content in Fritillaria ussuriensis Maxim. from different regions.

Figure 2

Related targets of lung cancer and peiminine. (A) The volcano map of differentially expressed genes. (B) Gene expression heat map. (C) PPI network of top 100 differentially expressed genes. (D) Venn diagram. (E) KEGG pathway. (F) GO analysis. (G) GSEA of the PI3K–Akt signaling pathway in lung cancer.

Figure 3

The PPI network. (A) A total of 105 targets. (B) PPI network of the 36 core targets. (C–I) Seven modules selected from the PPI network.

Figure 4

The 3D interaction between peiminine and the three genes. (A) PIK3CG; (B) SRC; (C) JAK3.

Figure 5

Molecular dynamics simulation of three complexes (JAK3 in blue, PIK3CG in orange, and SRC in red). (A–C) The RMSD curves of JAK3, PIK3CG, and SRC proteins with peiminine. (D–F) The RMSF curves of the complexes of JAK3, PIK3CG, and SRC proteins with peiminine. (G–I) The Rg curves of the complexes of JAK3, PIK3CG, and SRC proteins with peiminine. (J–L) The fluctuation curve of the number of hydrogen bonds formed between the JAK3, PIK3CG, and SRC proteins with peiminine. (M–O) The SASA curves of the complexes of the JAK3, PIK3CG, and SRC proteins with peiminine.

Figure 6

Molecular dynamics simulation. (A–C) Free energy distribution diagrams of the JAK3, PIK3CG, and SRC complexes with peiminine. (D–F) Structural comparisons at 0, 25, 50, 75, and 100 ns show peiminine’s positions (red, green, blue, yellow, and orange, respectively). (G–I) Average binding free energies of the complexes, calculated using MM/GBSA. Key terms include Van der Waals (VDWAALS), electrostatic (EEL), polar solvation (EGB), non-polar solvation (ESURF), molecular mechanics (GGAS), solvation (GSOLV), and total binding free energy (TOTAL). (J–L) Energy contributions of key residues in the JAK3, PIK3CG, and SRC proteins binding to peiminine. Critical binding contributions—JAK3: LEU-970 (−2.08 kcal/mol), PHE-833 (−2.07 kcal/mol), and ARG-948 (−1.70 kcal/mol); PIK3CG: ARG-284 (−1.38 kcal/mol), ARG-849 (−1.16 kcal/mol), and ARG-277 (−1.10 kcal/mol); SRC: ILE-153 (−2.72 kcal/mol).

Figure 7

Survival, expression, and correlation analysis. (A) Kaplan–Meier survival curve of AKT2. (B) Kaplan–Meier survival curve of PRKCA. (C) Correlation analysis of PIK3CG, SRC, JAK3, AKT2, and PRKCA. * for p value less than 0.05, ** for p value less than 0.01, and *** for p value less than 0.001.

Figure 8

The effect of peiminine in H1299 cells. (A) Cell viability of H1299 cells with different doses of peiminine. (B–F) The transcription levels of key genes after peiminine administration. Orange for untreated group, blue for 6 μM peiminine treatment group, green for 12 μM peiminine treatment group, and red for 25 μM peiminine treatment group. (B) PI3K. (C) PTEN. (D) Bax. (E) Bcl-2. (F) AKT. * p < 0.05 and ** p < 0.01 versus group without peiminine treatment.