Effect of Helix aspersa extract on TNFα, NF-κB and some tumor suppressor genes in breast cancer cell line Hs578T

BACKGROUND The garden snail, Helix aspersa, is a big land snail widely found in the Mediterranean countries. It is one of the most consumed species and widely used in zootherapy. OBJECTIVE The present study was carried out to investigate for the first time the first time the antitumor activity of an aqueous extract from Helix aspersa. MATERIALS AND METHODS The effect of H. aspersa extract was studied on a triple negative breast cancer cell line Hs578T. Firstly, the morphological changes and the mode of cell death induced by the extract have been evaluated by microscopy and acridine orange/ethidium bromide staining. The effect of the extract at dilution 0.1% and 1% was then tested on some genes, regulators of cell death and proliferation like tumor necrosis factor α (TNFα), NF- κB, and the tumor suppressor genes P53 and PTEN. RESULTS Data demonstrate that the extract induces necrosis in tumor cells. It enhances significantly the expression of TNFα; mRNA levels were 20 and 10 times more important in treated cells compared to nontreated cells. NF-κB and PTEN were inhibited with the dilution 1% after 8 and 24 hours of treatment. P53 expression was further inhibited but only with the highest dose, after 4, 8, and 24 hours. CONCLUSION Our results show that H. aspersa extract has an antitumor activity against Hs578T cells; it is a potent stimulator for TNFα and a good inhibitor for NF-κB. Abbreviations used: AO: acridine orange; Bcl-2: B cell lymphoma 2. cDNA: complementary DNA; ELISA: enzyme linked immunosorbent assay; EB: ethidium bromide; IC50: the half maximal inhibitory concentration; mRNA: messenger RNA. MAPK: mitogen-activated protein kinase; NF-κB: nuclearfactorkappa B; PBS: phosphate buffered saline. PI3K: phospho-inositol 3 kinase; PTEN: phosphatase and tensin homolog; ROS: reactive oxygen species. RT-PCR: reverse transcription polymerase chain reaction; TNFα: tumor necrosis factor alpha. TNFR1: TNF receptor-1; TP53: tumor protein 53.