Abstract
Purpose : To investigate the in vitro anti-proliferative effect and mechanism of action of Moringa oleifera Lam. leaf extract on human colon carcinoma HCT116 cell line. Methods : M. oleifera leaves were extracted with methanol. It was fractionated by Sephadex LH-20 column chromatography. Several fractions were identified by thin layer chromatography (TLC), proton nuclear magnetic resonance (1H NMR) and mass spectrometry (MS). The growth inhibitory activity and mechanism of action of the extracts in HCT116 colon cancer cells were investigated by 3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay and Western blotting. Results : Successive fractions from M. oleifera leaf crude extracts by column chromatography were combined into four pooled batches (MOL1 - MOL4) according to their absorbance at 260 nm and TLC pattern. MOL2 and MOL3 contain astragalin and isoquercetin, respectively. The results from MTT assay indicated that cell proliferation was significantly (p < 0.05) inhibited in a concentration-dependent fashion, especially by MOL2, MOL3 and MOL4. MOL2 and MOL3 exhibited a stronger cell growth inhibition than their major ingredients. The anti-proliferative activity of MOL2 - MOL4 in HCT116 colon cancer cells was mediated by downregulation of ERK1/2 phosphorylation. Conclusion : M. oleifera leaf extract has a strong anti-proliferative activity which is exerted by decreasing ERK1/2 phosphorylation. Thus, the extract has a potential for use in cancer chemoprevention. Keywords : Moringa oleifera , Anti-proliferation, Colon cancer, AKT, ERK1/2 phosphorylation, Chemoprevention
Highlights
Cancer is one of the leading diseases that results in hundreds of thousand deaths among Thais [13]
Our preliminary results suggested that the crude extract from M. oleifera leaves shows inhibitory effect on cell growth in HCT 116 cells
Our results showed that HCT116 treated with M. oleifera leaf extracts resulted in drastically decreased p-ERK1/2 and little decreased AKT expression, but not p38 mitogen-activated protein kinases (MAPKs) in a concentration dependent manner
Summary
Cancer is one of the leading diseases that results in hundreds of thousand deaths among Thais [13]. M. oleifera leaf extracts inhibit human epidermal carcinoma (KB) cell growth analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenytetrazolium bromide (MTT) assay [14]. AKT and ERK signaling pathways have been shown to coregulate several proteins promoting cancer cell survival and tumor growth [20]. A previous study showed that bioactive fractions from column chromatography separation of M. oleifera leaf extract exhibit apoptotic induction activity in HCT 116 colon cancer cells [21]. Dried methanol extract of M. oleifera leaves was subjected to gel filtration on Sephadex LH-20 with four pooled fractions (MOL1 - MOL4) collected. Varying concentrations of the M. oleifera pooled fractions were added into the cells for 48 h in triplicate cultures, compared with positive and negative controls.
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