Objective:

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To explore the key target molecules and potential mechanisms of oridonin against non-small cell lung cancer (NSCLC).

Methods:

2

The target molecules of oridonin were retrieved from Similarity Ensemble Approach (SEA)

Search Tool for Interacting Chemicals (STITCH)

SuperPred and TargetPred databases; target genes associated with the treatment of NSCLC were retrieved from GeneCards

DisGeNET and TTD databases. Then

the overlapping target molecules between the drug and the disease were identified. The protein–protein interaction (PPI) was constructed using the STRING database according to overlapping targets

and Cytoscape was used to screen for key targets. Molecular docking verification were performed using AutoDockTools and PyMOL software. Using the DAVID database

Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were conducted. The impact of oridonin on the proliferation and apoptosis of NSCLC cells was assessed using cell counting kit-8

cell proliferation EdU image kit

and Annexin V-FITC/PI apoptosis kit respectively. Moreover

real-time quantitative PCR and Western blot were used to verify the potential mechanisms.

Results:

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Fifty-six target molecules and 12 key target molecules of oridonin involved in NSCLC treatment were identified

including tumor protein 53 (TP53)

Caspase-3

signal transducer and activator of transcription 3 (STAT3)

mitogen-activated protein kinase kinase 8 (MAPK8)

and mammalian target of rapamycin (mTOR). Molecular docking showed that oridonin and its key target molecules bind spontaneously. GO and KEGG enrichment analyses revealed cancer

apoptosis

phosphoinosit

ide-3 kinase/protein kinase B (PI3K/Akt)

and other signaling pathways.

In vitro

experiments showed that oridonin inhibited the proliferation

induced apoptosis

downregulated the expression of Bcl-2 and Akt

and upregulated the expression of Caspase-3.

Conclusion:

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Oridonin can act on multiple targets and pathways to exert its inhibitory effects on NSCLC

and its mechanism may be related to upregulating the expression of Caspase-3 and downregulating the expressions of Akt and Bcl-2.