HPTLC estimation and anticancer potential of Aloe perryi petroleum ether extract (APPeE): A mechanistic study on human breast cancer cells (MDA-MB-231)

Abstract

Objectives: Aloe perryi plant in the genus Aloe (family: Asphodelaceae), have been used in traditional medicine systems. Herein, we aimed to analyze the anticancer effects of A. perryi petroleum ether extract (APPeE) against human breast cancer (MDA-MB-231) and normal (HEK-293) cell lines. Further the biomarker stigmasterol, isolated from A. perryi extract was quantified by a densitometric high-performance thin layer chromatography (HPTLC) method. Methods: The cytotoxic potential of APPeE was measured by MTT assay, neutral red uptake (NRU) assay, and morphological identification. Results: APPeE-induced a concentration dependent strong cytotoxic effects on MDA-MB-231 cells with an IC50 value of 24.5 μg/ml in contrast to HEK-293 (IC50 value of > 100 μg/ml). Similar results were obtained by NRU assay. Further cytotoxic concentrations of APPeE increased reactive oxygen species (ROS) production and activated caspase-3 and −9, leading to apoptosis in MDA-MB-231 cells. Additionally, a significant augment in the expression of p53, Bax, caspase-3 and −9 with a decline in Bcl-2 gene expression confirmed the involvement of apoptosis pathway in MDA-MB-231 cell death. In HPTLC analysis, the solvent system hexane: ethylacetate (8:2 v/v) furnished a sharp peak for stigmasterol (Rf = 0.19 ± 0.001). The low value of % relative standard deviation (RSD) (0.97–1.39) proposed the method is robust. The limit of detection (LOD) and limit of quantification (LOQ) were established as 13.82 and 41.89 ng, respectively. The stigmasterol in APPeE was quantified as 0.238% w/w of dried APPeE. Conclusions: Hence, this study concludes the promising anticancer effect of APPeE, which might be endorsed to the existence of known anticancer biomarker stigmasterol.

Keywords

• Aloe perryi
• Anticancer activity
• Cytotoxicity
• ROS generation
• Caspase activity
• Apoptosis