Effects of spider Macrothele raven venom on cell proliferation and cytotoxicity in HeLa cells
Abstract
Aim:To examine the effect of venom from the spider Macrothele raven on cell
proliferation and cytotoxicity in human cervical carcinoma, HeLa cells. Methods:
Morphological and biochemical signs of apoptosis appeared using acridine orange-
ethidium bromide (AO/EB) staining. Marked morphological changes in HeLa
cells after treatment with spider venom were observed using scanning electron
microscopy (SEM) and transmission electron microscopy (TEM). Cell proliferation
and cytotoxicity were determined by [methyl-3H] thymidine assay ([3H]TdR)
and lactate dehydrogenase (LDH) release, respectively. DNA fragmentation and
cell cycle distribution were monitored using flow cytometry. In addition, Western
blot analysis was used to evaluate the level of caspase-3 expression. In vivo
examination of the inhibition of the size of tumors in nude mice treated with spider
venom was measured. Results: Marked morphological changes were observed
using AO/EB staining, SEM and TEM assay. Spider venom at concentrations of
10–40 mg/L caused dose- and time-dependent inhibition of HeLa cell proliferation.
The ratio of apoptosis and necrosis increased. The activity of caspase-3 was
upregulated after spider venom treatment. In vivo study of tumor size revealed
that tumors significantly decreased in size from controls to tumors treated for 3
weeks with spider venom (P<0.05). Conclusion: The inhibition of HeLa cells by
the venom of the spider Macrothele raveni was carried out in three ways: induction
of apoptosis, necrosis of toxicity damage and direct lysis. Spider venom is a
novel anti-tumor material both in vitro and in vivo.
Keywords:
proliferation and cytotoxicity in human cervical carcinoma, HeLa cells. Methods:
Morphological and biochemical signs of apoptosis appeared using acridine orange-
ethidium bromide (AO/EB) staining. Marked morphological changes in HeLa
cells after treatment with spider venom were observed using scanning electron
microscopy (SEM) and transmission electron microscopy (TEM). Cell proliferation
and cytotoxicity were determined by [methyl-3H] thymidine assay ([3H]TdR)
and lactate dehydrogenase (LDH) release, respectively. DNA fragmentation and
cell cycle distribution were monitored using flow cytometry. In addition, Western
blot analysis was used to evaluate the level of caspase-3 expression. In vivo
examination of the inhibition of the size of tumors in nude mice treated with spider
venom was measured. Results: Marked morphological changes were observed
using AO/EB staining, SEM and TEM assay. Spider venom at concentrations of
10–40 mg/L caused dose- and time-dependent inhibition of HeLa cell proliferation.
The ratio of apoptosis and necrosis increased. The activity of caspase-3 was
upregulated after spider venom treatment. In vivo study of tumor size revealed
that tumors significantly decreased in size from controls to tumors treated for 3
weeks with spider venom (P<0.05). Conclusion: The inhibition of HeLa cells by
the venom of the spider Macrothele raveni was carried out in three ways: induction
of apoptosis, necrosis of toxicity damage and direct lysis. Spider venom is a
novel anti-tumor material both in vitro and in vivo.