Proceedings of The Physiological Society
University of Oxford (2011) Proc Physiol Soc 23, PC173
Cellular mechanism(s) of action of a water soluble extract of M. Charantia in inducing cell death on different cancer cell lines.
G. Manoharan1, J. Singh1
1. School of Pharmacy and Biomedical Sciences, University of Central Lancashire, Preston, United Kingdom.
A multitude of plants have been used extensively for the treatment of cancers throughout the world. Much research has been focused on the scientific evaluation of several photochemical from the tropical plant Momordica charantia (M charantia) which has been used frequently as an anti-cancer agent. In a previous communication (Gunasekar et al, 2010), we have shown that the crude water soluble extract of M. charantia can evoke both time and dose dependent effects on cancer cell death. This study now investigated the cellular mechanism(s) whereby the crude water soluble extract of M. charantia can elicit cell death employing 1321N1, Gos-3, U87-MG, Weri Rb1, Sk Mel, Corl -23 compared to normal healthy L6 muscle cell line. This study measured the release of cytochrome-c, the activities of caspase-3 and caspase-9 and intracellular free calcium concentrations [Ca 2+]i in the different cell lines following stimulation with 800 µg/ml of the crude water soluble extract of M. charantia for 24 hrs. Initial results have shown that the crude water soluble extract of M. Charantia can evoke significant (Student’s t-test; p<0.05) increases in the activities of both caspase-3 and caspase-9 compared to untreated cell lines over the same period. Typically, mean (±SD) values for caspase-3 activity (pmol AMC min1(mg protein1) were 0.42 ± 0.21, 0.37±0.17, 0.41±0.26, 0.51±0.32, 0.64±0.37, 0.67±0.43, 0.34±0.21, n = 12 in untreated cell lines compared to 0.84±0.42, 0.79±0.36, 0.77±0.41, 0.86±0.39, 0.95±0.49, 0.83±0.47, 0.62±0.36, n = 12 in the following treated cell lines 1321N1, Gos-3, U87-MG, Weri Rb1, Sk Mel and Corl -23, respectively. The activity was much smaller in L6 cell lines. Similarly, the crude water soluble extract of M. Charantia can elicit significant (p<0.05) increases in cytochrome-c release in the six different cancer cell lines compared to untreated cells. The results also show that the crude water-soluble extract of M. Charantia (800 µg) can evoke significant (p<0.05) and time-dependent increases in [Ca2+]i in all the six cancer cell lines employed in this study over a duration of 420 min compared to basal [Ca2+]i at the start of the experiment (0 min) and [Ca2+]i in each respective untreated cell line incubated alone in the medium for 420 min. The results also show that the extract had little or no significant effect on L6 skeletal muscle cell line. Mean (±SD) basal [Ca2+]i in this series of experiment was 0.17 ± 0.09 ratio units (intensity), n = 82. The results indicate that the crude extract of M. charantia can exert its anti-cancer effect (cell death) on cancer cells via damage of cell mitochondria body resulting in elevation in such cellular mediators as [Ca2+]I, caspase-3 and caspase-9 and release of cytochrome-c.
Where applicable, experiments conform with Society ethical requirements