Metabolomic and biochemometric profiling of Tecoma stans with mechanistic insights of its in vitro cytotoxic activity.

Tecoma stans is an ornamental plant recognized for its diverse biological activities, including notable cytotoxic effects. This study integrates LC-MS/MS-guided biochemometric analysis with mechanistic cytotoxicity evaluation to prioritize metabolites potentially associated with the cytotoxic activity of T. stans leaves. The hydroalcoholic extract and its solvent fractions n-hexane (HEX), dichloromethane (DCM), and ethyl acetate (EAC) were assessed for in vitro cytotoxicity against human lung adenocarcinoma (A549) and ovarian carcinoma (SKOV-3) cell lines using the MTT assay. Comprehensive LC-MS/MS-based metabolomic profiling and biochemometric correlation analysis were performed on the DCM fraction. The major correlated compound, chrysoeriol (CRY), was isolated and, along with the DCM fraction, subjected to cell cycle analysis, apoptosis assays, scratch wound healing assays, and ELISA-based quantification of apoptotic and metastatic markers (caspase-3, p-STAT3, Bcl-2, and MMP-2) in SKOV-3 cells. The DCM fraction exhibited the highest potency, particularly against SKOV-3 cells (IC₅₀ = 18.50 µg/mL). Metabolomic profiling led to the annotation of 107 metabolites, where CRY, α-sulfoquinovosyl monoacylglyceride (α-SQMG), tecomanine, and hydroxyskytanthine were prioritized as metabolites potentially associated with cytotoxic activity. DCM induced pronounced sub-G₁ arrest (71.42%, P < 0.0005) and increased late apoptosis to 23.32%, whereas CRY elevated sub-G₁ accumulation to 48.02% and late apoptosis to 12.22% (P < 0.0005). Furthermore, both treatments markedly reduced scratch wound closure and modulated key signaling pathways through caspase-3 activation and suppression of p-STAT3, Bcl-2, and MMP-2. The CRY and DCM fraction of T. stans leaves exerted their cytotoxic effects against SKOV-3 cells through the induction of apoptosis, wound closure inhibition and modulation of key oncogenic proteins. This study provides the first biochemometric and mechanistic validation of T. stans leaves DCM fraction and CRY in SKOV-3 cells, highlighting the potential combined contribution of its phytochemical constituents.