The hemolymph-derived
achatinin(H) (
lectin) from Achatina fulica showed a marked cytotoxic effect on MCF7, a
human mammary carcinoma cell line. IC(50) values as measured by the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium
bromide assay for
achatinin(H) ranged from 6 to 10 microg/ml in the MCF7 cells. MCF7 cells showed significant morphological changes leading to cell death. The above cell death was observed after 48 h of treatment with 8 microg/ml when compared to untreated cells. Alterations in the
tumor marker enzymes, as well as in
antioxidant enzymes, were observed after
achatinin(H) treatment. The specificity and purity of the
achatinin(H) was confirmed by the Western blot assay.
Achatinin(H) binding to MCF7 cells was detected by anti-
achatinin(H), and visualization of the
achatinin(H) binding sites on confluent MCF7 cells was confirmed by flourescein
isothiocyanate conjugated secondary antibody. MCF7-treated cells fluoresced, indicating the presence of
achatinin(H) binding sites. Fluorescence-activated cell sorting analysis of the cell cycle showed a significant increase in S-phase in MCF7 cells after 48 h of
achatinin(H) treatment. The cells were arrested in G(2)/M phase of the cell cycle after 48 h with significant changes in cell viability. Cellular damage was confirmed by
agarose gel electrophoresis with the characteristic appearance of
a DNA streak in treated MCF7 cells indicating the ongoing apoptosis.