Objective:

To evaluate the therapeutic efficacy of exosome-like nanoparticles derived from Nerium oleander flowers (NELNs) against triple-negative breast cancer (TNBC) in vitro.

Approach:

• Cytotoxicity Assessment: Cytotoxicity of NELNs was evaluated using the MTT assay.
• Apoptosis and ROS Generation Analysis: Apoptosis and reactive oxygen species (ROS) generation were examined using AO/EtBr and DCFDA staining through fluorescence microscopy.
• Flow Cytometry: Flow cytometry was employed for confirming apoptosis and analyzing cell cycle progression.
• Network Pharmacology: Network pharmacology was used for target prediction, PPI network construction, and functional enrichment analysis, followed by molecular docking of NELN-encapsulated metabolites.

Key Findings:

• NELNs induced significant cytotoxicity in MDA-MB-231 cells.
• NELNs promoted apoptosis and increased intracellular ROS levels.
• Flow cytometry confirmed apoptotic induction and cell cycle alterations.
• Pathway enrichment analysis highlighted arachidonic acid and linoleic acid metabolism.
• Cytochrome P450 enzymes (CYP1A2, CYP2C9, CYP3A4) were identified as putative targets.

Interpretation:

The study provides preliminary evidence that NELNs exert antiproliferative and pro-apoptotic activity against TNBC cells, with insights into potential mechanisms.

Limitations:

• The study is limited to in vitro conditions and may not fully represent in vivo efficacy.
• Further validation in animal models and clinical studies is necessary.

Conclusion:

Further investigation of NELNs is warranted based on the findings.