Cytotoxic and apoptotic effects of scorpion Leiurus quinquestriatus venom on 293T and C2C12 eukaryotic cell lines

Abstract

Scorpion venom toxicity is of major concern due to its influence on human activities and public health. The cytotoxicity and apoptosis induced by scorpion L. quinquestriatus venom on two established eukaryotic cell lines (293T and C2C12) were analyzed. Both cultured cell lines were incubated with varying doses (10, 20, and 50 µg/ml) of scorpion venom in serum free medium (SFM) for 0.5, 1, 2, 4, and 8 hours at 37°C. The percentage of total lactate dehydrogenase (LDH) released in the culture during venom incubation was used as an index of cell damage. Control culture was treated with an equal amount of SFM. Cell injury was recognized morphologically and apoptosis was researched by a Fluorescing Apoptosis Detection System using the principle of TUNEL (TdT-mediated dUTP Nick-End Labelling) assay and confirmed by another assay concerning nuclear DNA staining with DAPI stain. Cytotoxicity was remarkable and cell survival highly reduced at the highest tested concentration (50 µg/ml). These effects were rapid and observed within 30 minutes. The apparent initial damage to the nucleus and lysis of the plasmalemma and/or organelle membranes, which was evident by a significant increase in cytosolic LDH release, suggested that this toxin acts at the membrane level. The morphological changes that occurred in apoptotic cells include condensation and compartmentalization of nuclear and cytoplasmic materials into structurally preserved membrane-bound fragments or blebs. The cytotoxic effects are dose and time dependent and cell death by apoptosis was more characteristic of 293T cells than C2C12 cells. The apoptotic effects were more prominent and clear in the early stages of toxicity, while other forms of cell damage such as swelling, rupture, and/or necrosis occurred at later stages

Resistance of cervical adenocarcinoma cells (HeLa) to venom from the scorpion Centruroides limpidus limpidus

“…MTS metabolism assays detected no venom-related effects on the viability of HeLa cells (Figure 3 ). These results contrast with what has been reported by others, specifically regarding the effects of crude venom of different scorpion species on several cancer cell lines, where cell viability is affected in a dose-dependent manner [ 4 , 14 , 15 ]. However, we found that crude Cll venom was toxic to Balb/c mice macrophages, as previously described in a Tityus serrulatus scorpion venom challenge of Balb/c mice macrophages [ 28 ].…”

New Trends in Cancer Therapy and Antiviral Drug Research.

“…It was also reported that atroporin and kaotree derived from Crotalusatrox and Naja n. kaouthia venoms had anti-proliferative effect to some human breast, colon, liver, and ovary as well as animal cancer cell lines at concentrations as low as 0.5 μg/ml, in addition to their synergetic activity as anti-proliferating to cancer cells but not to normal cells (Lipps, 1999 In agreement with results obtained in this study it was found that venom inducedcytotoxic effect may be mediated by alterations in apoptotic gene regulation (Torii et al, 1997;Abe et al, 1998;Reinaldo et al, 2002;Correa et al, 2002). Moreover, L-amino acid oxidase enzyme associated apoptotic processes may be involved in the growth inhibition of the tumor cells (Raibekas and Massey, 1998).Another study demonstrated thatLeiurusquinquestriatusscorpion venom exhibited apoptotic effect on 293T and C2C12 cell lines in different concentrations (Omran, 2003).Snake venom may also inhibit cancer progression in laboratory animals (Zhou et al, 2000;Hantgan et al, 2005).Cytotoxicity records were also supported on small cell lung cancer cells (SCLC) (Tang et al, 2003;Poulsen et al, 2005).…”

Comprehensive analysis of venom from the scorpion Centruroides tecomanus reveals compounds with antimicrobial, cytotoxic, and insecticidal activities

“…Our results are consistent with studies reported by Gupta et al (2010) and Zargan et al (2011), who proved that 50 and 100 mg of venom from the Iranian scorpion Odontobuthus doriae, assayed on breast cells (line MCF-7) and neuroblastoma cells (line SH-SY5Y) causes changes in cell morphology and induces apoptosis. Similarly, Omran (2003), using 50 mg of soluble venom from the Egyptian scorpion Leiurus quinquestriatus showed the presence of cellular damage and apoptosis. In our case, the soluble venom of C. tecomanus tested on lymphoma cells (L5178Y) at concentration of 100 mg/mL causes more than 80% cell death; yet, at lower concentration of 1 mg/mL a 20% cytotoxicity was observed.…”