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Selected compounds isolated from Morinda citrifolia modulate prostate cancer cell growth

Published online by Cambridge University Press:  04 June 2010

V. Nitteranon ,
J. Cai ,
J. Xu ,
R. G. Godbee ,
K. L. Parkin  and
B. J. Darien
V. Nitteranon
Affiliation:
Department of Food Science, College of Agricultural and Life Sciences, Madison, WI, USA
J. Cai
Affiliation:
Department of Medical Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI, USA
J. Xu
Affiliation:
Department of Medical Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI, USA
R. G. Godbee
Affiliation:
Department of Animal Biotechnology/Veterinary Medicine, University Nevada, Reno, NV, USA
K. L. Parkin
Affiliation:
Department of Food Science, College of Agricultural and Life Sciences, Madison, WI, USA
B. J. Darien
Affiliation:
Department of Food Science, College of Agricultural and Life Sciences, Madison, WI, USA
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Abstract

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Proceedings of the Nutrition Society , Volume 69 , Issue OCE3: 3rd Immunonutrition Workshop, 21–24 October 2009, Girona, Spain , 2010 , E232
Copyright
Copyright © The Authors 2009

Morinda citrifolia (noni) fruit is a well-recognised natural product that reportedly has a broad range of immune enhancing effects(Reference Pawlus and Kinghorn1). Although several classes of metabolites have been described, few studies have evaluated the effects of compounds isolated from noni fruit on immune function(Reference Potterat and Hamburger2). Of particular interests were two classes of compounds, coumarins and flavanoids. Scopoletin is a pharmacologically active coumarin compound, which has been isolated from noni fruit. Reagent grade scopoletin inhibits the production of inflammatory cytokines. Flavanoids, including rutin, which is in noni, and quercetin, which is the aglycon form of rutin, have recognised antioxidant, anti-inflammatory and antineoplastic properties. The objective of this study was to investigate the dose response of two extraction fractions from noni puree, containing scopoletin and flavanoid compounds, on nitric oxide induction in RAW cells and cell viability in prostatic cancer (PC3) cells. Cell viability was quantified by Calcein AM, MTT and propidium iodide (PI) staining assays. Noni powder was extracted with ethyl acetate (EA) for compound isolation. The crude extract was fractionated using flash column chromatography with 30% acetone/hexane to obtain four fractions and F3 was identified as scopoletin. Fraction F4 was fractionated further using reversed-phase TLC with 50% acetone/water to obtain 5 fractions (F4.1, F4.3, F4.4, F4.5 and F4.6). F4.4 was identified as flavonoid-like compound (Fig. 1)

Fig. 1. NO production. Dose response of LPS-induced NO production to F3 and F4.4 fraction in RAW cells. F3 (scopoletin) IC50=72.4 μg/ml; F4.4 (Flavanoid) IC50=79 μg/ml. F3 decreased the production of nitrite in RAW cells when activated with LPS.F4.4 exhibited anti-inflammatory activity in LPS-stimulated RAW cells.

Fig. 2. Effects of different concentrations of noni EA, F3 (scopoletin rich) and reagent scopoletin on PC3 viability (MTT). No cytotoxicity was observed below 100 μg/ml.

Figures 2–4: PC3 cell viability.

Fig. 3. Effects of different concentrations of noni EA, F3 (scopoletin rich) and reagent scopoletin on PC3 viability (Calcein AM). The EA fraction inhibited PC3 cell growth by about 30% across all concentrations, while F3 showed greater inhibition (45%) at 200 μg/ml.

Fig. 4. Effects of different concentrations of noni EA on PC3 viability (PI). Inset shows individual concentration effects on PI uptake by flow cytometry and graphed below. Cell death >200 μg/ml.

These preliminary studies demonstrate that scopoletin and flavanoid rich fractions from noni puree inhibited NO production and PC3 cell growth. Further investigations of the mechanism by which different noni compounds singularly or in combination modulate cancer growth is underway.

References

1.Pawlus, AD & Kinghorn, AD (2007) J Pharm Pharmacol 59, 1587–1609.CrossRefGoogle Scholar
2.Potterat, O & Hamburger, M (2007) Planta Med 73, 191–199.CrossRefGoogle Scholar
Figure 0

Fig. 1. NO production. Dose response of LPS-induced NO production to F3 and F4.4 fraction in RAW cells. F3 (scopoletin) IC50=72.4 μg/ml; F4.4 (Flavanoid) IC50=79 μg/ml. F3 decreased the production of nitrite in RAW cells when activated with LPS.F4.4 exhibited anti-inflammatory activity in LPS-stimulated RAW cells.

Figure 1

Fig. 2. Effects of different concentrations of noni EA, F3 (scopoletin rich) and reagent scopoletin on PC3 viability (MTT). No cytotoxicity was observed below 100 μg/ml.

Figure 2

Fig. 3. Effects of different concentrations of noni EA, F3 (scopoletin rich) and reagent scopoletin on PC3 viability (Calcein AM). The EA fraction inhibited PC3 cell growth by about 30% across all concentrations, while F3 showed greater inhibition (45%) at 200 μg/ml.

Figure 3

Fig. 4. Effects of different concentrations of noni EA on PC3 viability (PI). Inset shows individual concentration effects on PI uptake by flow cytometry and graphed below. Cell death >200 μg/ml.