Abstract
An alkaloid compound from the hairy root culture of Eurycoma longifolia has been isolated and characterised as 9-methoxycanthin-6-one. The aims of these studies were to investigate the in vitro anti-cancer activities of 9-methoxycanthin-6-one against ovarian cancer (A2780, SKOV-3), breast cancer (MCF-7), colorectal cancer (HT29), skin cancer (A375) and cervical cancer (HeLa) cell lines by using a Sulphorhodamine B assay, and to evaluate the mechanisms of action of 9-methoxycanthin-6-one via the Hoechst 33342 assay and proteomics approach. The results had shown that 9-methoxycanthin-6-one gave IC50 values of 4.04 ± 0.36 µM, 5.80 ± 0.40 µM, 15.09 ± 0.99 µM, 3.79 ± 0.069 µM, 5.71 ± 0.20 µM and 4.30 ± 0.27 µM when tested in A2780, SKOV-3, MCF-7, HT-29, A375 and HeLa cell lines, respectively. It was found that 9-methoxycanthin-6-one induced apoptosis in a concentration dependent manner when analysed via the Hoechst 33342 assay. 9-methoxycanthine-6-one were found to affect the expressions of apoptotic-related proteins, that were proteins pyruvate kinase (PKM), annexin A2 (ANXA2), galectin 3 (LGAL3), heterogeneous nuclear ribonucleoprotein A1 (HNRNP1A1), peroxiredoxin 3 (PRDX3), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from the differential analysis of 2-DE profiles between treated and non-treated 9-methoxycanthine-6-one. Proteins such as acetyl-CoA acyltransferase 2 (ACAA2), aldehyde dehydrogenase 1 (ALDH1A1), capping protein (CAPG), eukaryotic translation elongation factor 1 (EEF1A1), malate dehydrogenase 2 (MDH2), purine nucleoside phosphorylase (PNP), and triosephosphate isomerase 1 (TPI1) were also identified to be associated with A2780 cell death induced by 9-methoxycanthine-6-one. These findings may provide a new insight on the mechanisms of action of 9-methoxycanthin-6-one in exerting its anti-cancer effects in vitro.
Highlights
Cancer cases are still on the rise in both developed and developing countries, and it is predicted that 14 million new cancer cases will emerge by 2035, representing almost 60% of the global cancer incidence, as compared to 6.7 million new cancer cases (47.5% of all cancers) reported in 2012 [1]
The 13CNMR peak assignments showed a methoxy carbon resonated at δ 56.2, a signal at δ 163.41 attributed to carbonyl carbon (C-6), and seven signals, δ 115.66 (C-1), δ 143.81 (C-2), δ 138.05 (C-4), δ 129.69 (C-5), δ 101.45 (C-8), δ 114.76 (C-10) and δ 123.95 (C-12), corresponding to seven aromatic carbons
Other studies have supported this finding in which the yield of 9-methoxycanthin-6-one (1) was higher in E. longifolia tissue cultures than in E. longifolia roots collected from the wild
Summary
Cancer cases are still on the rise in both developed and developing countries, and it is predicted that 14 million new cancer cases will emerge by 2035, representing almost 60% of the global cancer incidence, as compared to 6.7 million new cancer cases (47.5% of all cancers) reported in 2012 [1]. Cancer arises due to malfunctions of certain genes and their translated proteins that are related to the over cell proliferations, divisions and growth and/or suppressions of apoptosis-related proteins [3]. Apoptosis or programmed cell death is a process involving expressions of certain genes and proteins which signal the cells to commit suicide in order to eliminate the potential cancer cells’ development in our body [4]. Most of the cancer cells malfunction in apoptosis and many anti-cancer therapeutic agents were found to re-induce the apoptosis mechanism and produce a better outcome to prevent cancer cells’ proliferation [5]. Phytochemical(s) that enable the re-inducement of the apoptosis mechanism in the cancer cells may have the potential to be studied further towards development of anti-cancer therapeutic agents
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