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. 2009 Feb 11;57(3):826-31.
doi: 10.1021/jf803407q.

Eugenia jambolana Lam. berry extract inhibits growth and induces apoptosis of human breast cancer but not non-tumorigenic breast cells

Liya Li  1 , Lynn S Adams, Shiuan Chen, Caroline Killian, Aftab Ahmed, Navindra P Seeram
Affiliations

Eugenia jambolana Lam. berry extract inhibits growth and induces apoptosis of human breast cancer but not non-tumorigenic breast cells

Liya Li et al. J Agric Food Chem. .

Abstract

The ripe purple berries of the native Indian plant Eugenia jambolana Lam., known as Jamun, are popularly consumed and available in the United States in Florida and Hawaii. Despite the growing body of data on the chemopreventive potential of edible berry extracts, there is paucity of such data for Jamun fruit. Therefore our laboratory initiated the current study with the following objectives: (1) to prepare a standardized Jamun fruit extract (JFE) for biological studies and (2) to investigate the antiproliferative and pro-apoptotic effects of JFE in estrogen dependent/aromatase positive (MCF-7aro), and estrogen independent (MDA-MB-231) breast cancer cells, and in a normal/nontumorigenic (MCF-10A) breast cell line. JFE was standardized to anthocyanin content using the pH differential method, and individual anthocyanins were identified by high performance liquid chromatography with ultraviolet (HPLC-UV) and tandem mass spectrometry (LC-MS/MS) methods. JFE contained 3.5% anthocyanins (as cyanidin-3-glucoside equivalents) which occur as diglucosides of five anthocyanidins/aglycons: delphinidin, cyanidin, petunidin, peonidin and malvidin. In the proliferation assay, JFE was most effective against MCF-7aro (IC(50) = 27 microg/mL), followed by MDA-MB-231 (IC(50) = 40 microg/mL) breast cancer cells. Importantly, JFE exhibited only mild antiproliferative effects against the normal MCF-10A (IC(50) > 100 microg/mL) breast cells. Similarly, JFE (at 200 microg/mL) exhibited pro-apoptotic effects against the MCF-7aro (p <or= 0.05) and the MDA-MB-231 (p <or= 0.01) breast cancer cells, but not toward the normal MCF-10A breast cells. These studies suggest that JFE may have potential beneficial effects against breast cancer.

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Figures

Figure 1
Figure 1
Eugenia jambolana Lam. (Jamun) berries and seeds (picture by NS from fruits collected in St. Catherine, Jamaica, West Indies).
Figure 2A
Figure 2A
HPLC-UV chromatogram of Jamun Fruit Extract (JFE) at 520 nm. Anthocyanins were identified as delphinidin-diglucoside (peak 1, tR=22.4 min), cyanidin-diglucoside (peak 2, tR=25.4 min), petunidin-diglucoside (peak 3, tR=27.2), peonidin-diglucoside (peak 4, tR=30.7 min), and malvidin-diglucoside (peak 5, tR=31.9 min). Figure 2B: LC-MS/MS spectrum of the individual anthocyanins present in JFE identified as m/z M+;MS/MS ions as follows 2B1: delphinidin-diglucoside (627;465/303); 2B2: cyanidin-diglucoside (611;449/287); 2B3: petunidin-diglucoside (641;479/317), 2B4: peonidin-diglucoside (625;463/301), and 2B5: malvidin-diglucoside (655;493/331).
Figure 2A
Figure 2A
HPLC-UV chromatogram of Jamun Fruit Extract (JFE) at 520 nm. Anthocyanins were identified as delphinidin-diglucoside (peak 1, tR=22.4 min), cyanidin-diglucoside (peak 2, tR=25.4 min), petunidin-diglucoside (peak 3, tR=27.2), peonidin-diglucoside (peak 4, tR=30.7 min), and malvidin-diglucoside (peak 5, tR=31.9 min). Figure 2B: LC-MS/MS spectrum of the individual anthocyanins present in JFE identified as m/z M+;MS/MS ions as follows 2B1: delphinidin-diglucoside (627;465/303); 2B2: cyanidin-diglucoside (611;449/287); 2B3: petunidin-diglucoside (641;479/317), 2B4: peonidin-diglucoside (625;463/301), and 2B5: malvidin-diglucoside (655;493/331).
Figure 3
Figure 3
Antiproliferative activities of JFE against 3A: MCF10A (untransformed/non-tumorigenic) breast cells, 3B: MDA-MB-231 (estrogen independent) breast cancer cells and 3C: MCF7aro (estrogen dependent/aromatase positive) breast cancer cells. Cells were exposed to JFE at concentrations ranging from 0–100 µg/mL for 24, 48 or 72 h. Proliferation was measured via the CellTiter-Glo® Luminescent Cell Viability Assay. Data are expressed as percentage of untreated cells, mean ±SE and analyzed utilizing One-Way ANOVA followed by Dunnet’s Multiple Range test. Asterisk indicates a significant difference compared to untreated controls (p≤0.01).
Figure 4
Figure 4
Apoptosis in : MCF10A (untransformed/non-tumorigenic) breast cells, MCF-7aro (estrogen dependent/aromatase positive) and MDA-MB-231 cells (estrogen independent) breast cancer cells after 48 h of treatment with JFE. Data are expressed as a ratio of treated samples to media controls ± SEM and analyzed utilizing One-Way ANOVA followed by Dunnet’s Multiple Range test. Single asterisk indicates statistical significance from media controls p ≤0.05, double asterisk indicates statistical significance p ≤0.01.
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