The apoptotic mechanism is complex and involves many pathways. Defects can occur at any time along these pathways, resulting in malignant cell transformation and resistance to anticancer drugs. Collective efforts have made great progress in the implementation of natural products in clinical use and in discovering new therapeutic opportunities. This study aimed to screen volatile compounds of Warburgia salutaris leaf extracts and investigate their pro-apoptotic effects on MCF-7 cells. The approach was mainly based on determining cell viability using MTT and scratch assays, and DNA synthesis and damage using BrdU and comet assays, respectively. DAPI/PI stains were used for morphological analysis and expression was determined by RT-PCR and human apoptotic proteome profiler. Warburgia salutaris extracts exhibited antiproliferative effects on MCF-7 cells in a time- and dose-dependent manner. Acetone and methanol extracts exhibited low IC₅₀ at 24, 48 and 72 h. Furthermore, the scratch test revealed that MCF-7 does not metastasise when treated with IC₅₀. Expression showed upregulation of pro-apoptotic proteins and executioner caspases. Taken together, these findings suggest that leaves can promote apoptosis through the intrinsic apoptotic pathway, as observed by upregulation of the Bax and caspase 3 proteins. This paper provides new insights into the mechanisms of action of W. salutaris leaf extracts in the development of anticancer drugs.

Although medicinal plants possess vast biological properties, crude medicinal plant extracts often show limited therapeutic efficacy due to poor aqueous solubility, instability, and inadequate bioavailability, which restricts efficient intracellular delivery. As cancer is a genetic disease requiring intracellular and nuclear targeting, improved delivery systems are essential. Warburgia salutaris is traditionally used in Southern Africa and possesses reported anticancer and anti-inflammatory properties; however, its crude extracts exhibit suboptimal delivery characteristics. This study comparatively evaluated the anticancer effects of unencapsulated (WSN) and liposomal-encapsulated (WSE) crude leaf extracts, with emphasis on apoptotic mechanisms. Liposomal formulation was confirmed by FTIR, PXRD, and DLS, yielding stable nanoparticles (159.4 nm; PDI 0.114; +79.3 mV). Both WSN and WSE demonstrated efficacy, concentration-dependent cytotoxicity against MCF-7 breast cancer cells (IC50 < 0.0195 mg/mL) with minimal toxicity toward Vero kidney cells and Raw 264.7 macrophages. Mechanistically, WSN induced rapid cytotoxicity with necrotic features, whereas WSE promoted regulated apoptosis. Apoptosis was validated by DAPI/PI staining, Annexin V/PI flow cytometry, mRNA expression levels of Bax, Bcl-2, and caspase-3 measured with RT-PCR and proteome profiling array confirming activation of intrinsic and extrinsic pathways. Both extracts also reduced LPS-induced ROS production. LC-MS identified multiple bioactive phytochemicals. Overall, liposomal encapsulation enhanced therapeutic precision, stability, and selectivity cytotoxicity, supporting its development as a nanomedicine-based anticancer strategy.

Although medicinal plants possess vast biological properties, crude medicinal plant extracts often show limited therapeutic efficacy due to poor aqueous solubility, instability, and inadequate bioavailability, which restricts efficient intracellular delivery. As cancer is a genetic disease requiring intracellular and nuclear targeting, improved delivery systems are essential. Warburgia salutaris is traditionally used in Southern Africa and possesses reported anticancer and anti-inflammatory properties; however, its crude extracts exhibit suboptimal delivery characteristics. This study comparatively evaluated the anticancer effects of unencapsulated (WSN) and liposomal-encapsulated (WSE) crude leaf extracts, with emphasis on apoptotic mechanisms. Liposomal formulation was confirmed by FTIR, PXRD, and DLS, yielding stable nanoparticles (159.4 nm; PDI 0.114; +79.3 mV). Both WSN and WSE demonstrated efficacy and concentration-dependent cytotoxicity against MCF-7 breast cancer cells (IC₅₀ < 0.0195 mg/mL) with minimal toxicity toward Vero kidney cells and RAW 264.7 macrophages. Mechanistically, WSN induced rapid cytotoxicity with necrotic features, whereas WSE promoted regulated apoptosis. Apoptosis was validated by DAPI/PI staining, Annexin V/PI flow cytometry, mRNA expression levels of Bax, Bcl-2, and caspase-3 measured with RT-PCR and proteome profiling array, confirming activation of intrinsic and extrinsic pathways. Both extracts also reduced LPS-induced ROS production. LC-MS identified multiple bioactive phytochemicals. Overall, liposomal encapsulation enhanced therapeutic precision, stability, and selectivity cytotoxicity, supporting its development as a nanomedicine-based anticancer strategy.

Agapanthus africanus (L.) Hoffmanns. is a medicinal plant traditionally used in South Africa for its promise as a source of bioactive compounds with anticancer properties. This study aimed to investigate the apoptotic effects of A. africanus fractions on cancer cell lines and to identify the bioactive phytochemical constituents using gas chromatography-mass spectrometry analysis. To test for cytotoxicity, MCF-7, A549, and HeLa cancer cells were treated with crude extract, n-hexane, n-butanol, dichloromethane, and aqueous fractions of A. africanus extracts at different concentrations (0.00–1000 µg/mL). Total apoptosis was quantified using Annexin V/PI staining. The 4′,6-diamidino-2-phenylindole was used to detect nuclear morphological changes and the Caspase-GLO 3/7 assay was employed to check the caspase activation in the cancer cells. Expression of apoptosis-related (caspase-3, bax, bcl-2) genes was evaluated using real time-polymerase chain reaction. The crude extract of A. africanus exhibited dose-dependent cytotoxicity against MCF-7, A549, and HeLa cells, with IC₅₀ values of 130 µg/mL, 380 µg/mL, and <125 µg/mL, respectively. Among the tested fractions, the n-butanol fraction showed cytotoxicity towards MCF-7 cells with an IC₅₀ value of <870 µg/mL. In contrast, n-hexane, dichloromethane and the aqueous fractions exhibited higher IC₅₀ values against cancer cells. Flow cytometry analysis, which was applied to quantify total apoptosis, revealed that the crude extract of A. africanus induced apoptosis by (~60%) compared to the n-butanol fraction, which exhibited a moderate apoptotic effect (~27%). DAPI nuclear staining showed nuclear shrinkage and chromatin condensation in the MCF-7 cell line, whereas in Caspase-GLO 3/7, the crude extract and n-butanol fraction resulted in significant luminescence, indicating activation of caspase-3/7. Caspase-3/7 analysis showed A. africanus treatments produced varying levels of apoptotic activation. The crude extract increased caspase activity by 2.9-fold, while the n-butanol fraction induced a 1.7-fold rise compared with untreated cells. GC-MS chromatograms detected and identified 16 compounds in the fractionated n-butanol and 23 compounds from the crude extract of A. africanus. The major compounds identified from the n-butanol fraction included n-hexadecanoic acid; α-tocopherol and 9,12,15-octadecatrienoic acid, while the GC–MS profile of the crude extract was dominated by 6,10,14-trimethylpentadecan-2-one; 1,3,5-Triphenylcyclohexane and phytol. The study indicates the pro-apoptotic potential of A. africanus, particularly in its crude form, supporting its ethnopharmacological use and suggesting its relevance as a candidate for anticancer drug discovery.