Investigation of the Cytotoxic, Genotoxic, and Apoptosis-Inducing Effects of Estragole Isolated from Fennel (Foeniculum vulgare)

Abstract

The present study was undertaken to evaluate, in the HepG2 human hepatoma cell line, the in vitro cytotoxic, genotoxic, and apoptotic activities of estragole (1), contained in the essential oil of Foeniculum vulgare (fennel) and suspected to induce hepatic tumors in susceptible strains of mice. Toward this end, an MTT cytotoxicity assay, a trypan blue dye exclusion test, a double-staining (acridine orange and DAPI) fluorescence viability assay, a single-cell microgel-electrophoresis (comet) assay, a mitochondrial membrane potential (Δψm) assay, and a DNA fragmentation analysis were conducted. In terms of potential genotoxic effects, the comet assay indicated that estragole (1) was not able to induce DNA damage nor apoptosis under the experimental conditions used.

Cytotoxicity screening of essential oils in cancer cell lines

“…Despite the reports on the biological activities of these essential oils, data on their cytotoxicity are still scarce in the literature (Fabio et al, 2007;Gazim et al, 2014;Villarini et al, 2014).…”

Epilobium angustifolium L.: A medicinal plant with therapeutic properties

“…This cell line has been chosen for its high degree of morphological and functional differentiation in vitro, and also because it is a suitable model to study drug and plant metabolites targeting in vitro (34)(35). MTT assay has been used to test cell viability at different concentration of E. angustifolium extract (25,50,75,100,150,200, 300, 400 and 500 µg/mL). Incubation of E. angustifolium extract after 24h resulted in a reduction of proliferation in a concentrationdependent manner (Fig.…”

“…Concentration of cells was 4×10 4 for each well. Then, after 24 h, the medium was replaced with fresh complete medium, containing different concentrations (25,50,75,100,150,200, 300, 400, 500 μg/mL) of E. angustifolium extract and incubated for additional 24 h. MEM was used as a negative control and 1% solution of DMSO as positive control. After 24h, MTT reagent was dissolved in PBS 1X, and added to the culture at 0.5 mg/ml final concentration.…”

“…After incubation, samples underwent two washing steps and at the end of the second wash, samples were re-suspended in 250 mL of 1mg/mL of DAPI solution. Samples were analyzed with the NucleoCounter® NC-3000™ analysis system (25).…”

“…Ethanol was added to every tube and cell suspensions were kept at 4°C for 12 h. After centrifugation ethanol was removed, cell pellets were suspended in PBS and centrifuged again after 5 minutes. Then cell pellets were suspended in a solution containing 1 μg/ml DAPI, 0.1% triton X-100 in PBS, and load on A8-ChemoMetec TM slides and DNA fragmentation was investigated with NucleoCounter® NC-3000™ analysis system (25).…”