Evaluating the efficacy of an innovative herbal formulation (HF6) on different human cancer cell lines

Abstract

Cancer is reported to be the leading cause of death and illness worldwide. This research aims to evaluate the phytochemicals, antioxidant, cytotoxic, and apoptotic activities of the polyherbal formulation HF6. HF6 was prepared by blending equal quantities of plants powder, namely, Curcuma longa, Salvia officinalis, Cinnamomum zeylanicum, Capsicum annuum, Zingiber officinale, and Syzygium aromaticum, and later extracted using hexane (HF6H), chloroform (HF6C), ethyl acetate (HF6E), and methanol (HF6M) in Soxhlet apparatus. Among the four different extracts, only the hexane extract (HF6H) was significantly effective. The HF6H extract showed antioxidant and anticancer potentials against different cancer cell lines, and moderate cytotoxicity against non-cancer cells, rendering it a promising remedy. In addition, it exerted tremendous cytotoxic effects on MCF-7, Huh-7, HCT116, MDA-MB-231, LoVo, and HepG2 cells with IC₅₀ values of 2.02, 4.5, 6.9, 11.4, 23.5, and 34.7 µg/mL, respectively. The morphological hallmarks of apoptosis such as the rounding of cells, loss of contact with neighboring cells, formation of cell membrane blebbing, and microspike protrusion were detected using several different techniques. DAPI staining revealed apoptotic nuclear morphology such as condensation and DNA fragmentation. The morphological changes of MCF7 cells were also analyzed by AO/EB fluorescence staining. MCF7-stained green cells were viable cells, whereas the treated cells showed fragmented green nuclei representing early apoptosis. The phytochemical screening of HF6H showed positive results regarding the presence of alkaloids, polyphenols, flavonoids, and sterols. The GC-MS (gas chromatography-mass spectrometry) analysis of the HF6H extract indicated the presence of 12 compounds, mainly trans-caryophyllene (21.55%), cis-isoeugenol (18.42%), acetyleugenol (17.53%), alpha farnesene (10.0%), and zingiberene (8.55%). However, further investigation could be carried out to examine the toxicity of the extract on animal models.

Keywords: Anticancer; Antioxidant; Apoptosis; GC–MS; Polyherbal.