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Archive > Volume 32 Issue 8 > 2025,32(8):814-822. DOI:10.3872/j.issn.1007-385X.2025.08.004 Prev Next
Basic Research
Paeonol suppresses the proliferation, migration, and invasion abilities of lung cancer A549 cells via the EZH2/NXPH4/CDKN2A axis
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Abstract:
[Abstract] Objective: To investigate the effects of paeonol (Pae) on the proliferation, migration, and invasion abilities of lung cancer A549 cells by regulating the enhancer of zeste homolog 2/neurexophilin 4 /cyclin dependent kinase inhibitor 2A (EZH2/NXPH4/ CDKN2A) axis. Methods: A549 lung cancer cell lines were treated with Pae at gradient concentrations (0, 6.25, 12.5, 25, 50, 100, 200 μg/mL). CCK-8 assay was used to screen for suitable Pae concentrations. A549 cells were assigned into the control group (untreated A549 cells), the Pae group (12.5 μg/mL Pae), the Pae + siEZH2 group (transfected with siEZH2+12.5 μg/mL Pae), the Pae + siNC group (transfected with siNC+12.5 μg/mL Pae), the Pae + vectorNC group (transfected with vector NC + 12.5 μg/mL Pae), and the Pae + vectorEZH2 group (transfected with vector EZH2 + 12.5 μg/mL Pae). After the indicated treatments, the colony formation assay was applied to detect the number of cell clones. Cell cycle distribution was measured by flow cytometry. Transwell assay was used to detect changes in cell invasion ability. Scratch assay was employed to detect changes in cell migration ability. Flow cytometry was applied to detect cell apoptosis. WB assay was used to detect the protein expression levels of EZH2, NXPH4, CDKN2A, Bcl-2, and caspase-3. Only siEZH2 or only siNC were transfected into A549 cells. Cell apoptosis was measured by flow cytometry, and the expressions of Bcl-2 and caspase-3 proteins were detected by WB assay. AnA549-cell nude-mouse transplant model was established to evaluate the in vivo antitumor efficacy of Pae and its effects on the expressions of relevant proteins. Results: 12.5 μg/mL was chosen as the concentration for subsequent experiments. Compared with those in the control group, the cell clone number, the proportion of S phase cells, the migration ability, the invasion ability, and the expression levels of EZH2, NXPH4, and Bcl-2 proteins were significantly reduced in the Pae group, while the proportion of G0/G1 phase cells, the apoptosis rate and the expression levels of CDKN2A and caspase-3 proteins were significantly increased (all P < 0.05). Compared with those in the Pae + si NC group, the cell clone number, the proportion of S phase cells, the migration ability, the invasion ability, and the expression levels of EZH2, NXPH4, and Bcl-2 proteins decreased more markedly in the Pae + siEZH2 group, while the proportion of G0/G1 phase cells, the apoptosis rate and the expression levels of CDKN2A and caspase-3 proteins increased more markedly (all P < 0.05). Compared with those in the Pae+vector NC group, the cell clone number, the proportion of S phase cells, the migration and invasion abilities, and the expression levels of EZH2, NXPH4, and Bcl-2 proteins increased significantly in the Pae + vectorEZH2 group, while the proportion of G0/G1 phase cells, the apoptosis rate and the expression levels of CDKN2A and caspase-3 proteins decreased significantly (P < 0.05). After EZH2 knockdown, the apoptosis rate and caspase-3 expression of A549 cells increased significantly, while Bcl-2 expression decreased significantly (both P < 0.05). In vivo experiments showed that paeonol significantly reduced tumor volume and mass, and inhibited the activity of the EZH2/NXPH4/ CDKN2A pathway. Conclusion: Pae inhibits the proliferation, migration and invasion of A549 cells and induces apoptosis by down regulating the EZH2/NXPH4/CDKN2Apathway.
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