Evaluation of the effect of Nigella sativa extract on human hepatocellular adenocarcinoma cell line (HepG2) in vitro
© Khan et al; licensee BioMed Central Ltd. 2014
Published: 2 April 2014
Materials and methods
The whole extract of Nigella sativa (generously donated by the ENT research group, KAUH) was filter sterilized using 0.2µm syringe filters. The HepG2 cells were seeded at 3 x 104 cells/well of a 24-well tissue culture plate and cultured overnight in DMEM low glucose medium supplemented with 10% fetal bovine serum, 200mM GlutaMax, 1% penicillin/streptomycin under standard culture conditions of 37°C in a 5% CO2 air atmosphere. Following addition of fresh medium, Nigella sativa extract was added at various concentrations namely 0.1%, 0.3%, 0.5, 0.7%, and 1%; and the cells cultured for 24 h and 48 h. Nigella sativa extract was not added to the control wells. Changes in cell morphology was imaged using inverted phase contrast optics and the cell viability was assessed by MTT assay.
Control HepG2 cells maintained their typical morphology and formed a confluent monolayer. In contrast, the cells treated with Nigella sativa extract showed varying changes in morphology (cell shrinkage, membrane damage) resulting in cell death and gross decreases in cell numbers starting from 0.3% concentration at both 24 h and 48 h (Figure 1B). MTT assay demonstrated statistically significant decreases in cell proliferation with increasing concentrations of the drug at 24 h and 48 h. The mean decreases in cell proliferation were 18%, 42%, 54%, 56%, and 62% at 24hr; and 23%, 27%, 36%, 38% and 53% at 48hr for the concentrations 0.1%, 0.3%, 0.5%, 0.7% and 1% respectively (Figure 1C, 1D).
In the present study, the extract of Nigella sativa demonstrated inhibition of HepG2 cell line in vitro. Our results are in accordance with an earlier study  where a different form of Nigella sativa extract was found to inhibit the growth and proliferation of the HepG2. We therefore conclude that Nigella sativa extract has anticancer properties which needs further exploration and as such we are currently involved in identifying the active ingredient of the extract as well as the underlying molecular mechanism leading to cell death.
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