Anti-Cancer Activity of Manilkara Zapota Seed and Skin Extracts on Hela Cell Lines

Abstract

Objective: To assess the anticancer activity of ethanolic extracts of Manilkara zapota (MZ) seeds and skin against HeLa cell lines by Methyl Thiazolyl Tetrazolium bromide (MTT) and Lactate dehydrogenase (LDH) assay. Methodology: Crude extract of seeds and skin of MZ were prepared by standard procedures using ethanol. It was filtered and evaporated on water bath. The extract was dissolved in 65% ethanol, centrifuged at -4 degrees at 4000 rpm for 10 minutes and the supernatant was used. The anticancer activity of extracts was assessed using HeLa cancer cell lines by MTT and LDH assay. For MTT Assay, 96 well plate was seeded with 5000 HeLa cells per well, incubated at 37ºC under 5% CO2. After 24 hrs, cells were treated with the extracts (0.5-12 mg/ml for both skin and seed) and Paclitaxel (0.03-1mg/ml). After 48 hrs 100µl of 0.2% MTT and later 50µl of dimethyl sulfoxide (DMSO) were added and optical density was measured at 570nm. LDH Assay was done using the supernatant cell suspension and LDH reagent and absorbance read at 490nm. Results: The extracts were found to have anticancer activity and the IC-50 (Inhibitory Concentration - 50) of seed and skin extracts and Paclitaxel were 4µg/ml, 8µg/ml and 2 µg/ml respectively. Conclusion: The ethanolic extracts of MZ seeds and skin have shown anticancer activity against HeLa cells in vitro by MTT and LDH assay.