Antiproliferative activity of peels, pulps and seeds of 61 fruits

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Antiproliferative activity of peels, pulps and seeds of 61 fruits Fang Lia,1, Sha Lia,1, Hua-Bin Lia,*, Gui-Fang Denga, Wen-Hua Linga, Shan Wua, Xiang-Rong Xub,*, Feng Chenc a

Guangdong Provincial Key Laboratory of Food, Nutrition and Health, Department of Nutrition, School of Public Health, Sun Yat-Sen University, Guangzhou 510080, China b Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China c Institute for Food and Bioresource Engineering, College of Engineering, Peking University, Beijing 100871, China

A R T I C L E I N F O

A B S T R A C T

Article history:

Fruits are wildly consumed and considered to have various health benefits. The aim of this

Received 9 October 2012

study was to supply new information on the antiproliferative function of selected fruits for

Received in revised form

nutritionists and the general public. The in vitro antiproliferative activities of the peels, pulps

16 April 2013

and seeds of 61 fruits on four cancer cell lines, A549 (human lung cancer cells), MCF-7

Accepted 19 April 2013

(human breast cancer cells), HepG2 (human hepatoma cells) and HT-29 (human colon can-

Available online 20 May 2013

cer cells) were evaluated by MTT assay. The results revealed that different fruits and different parts of one fruit exhibited different antiproliferative capacities. Nine of the 162 samples

Keywords:

from 61 fruits showed remarkable inhibitory effects toward the four cancer cell lines, and all

Fruit

decreased the viability of the cancer cell lines in a dose-dependent manner. The results sug-

Peel

gested that some fruits (such as honey peach, salak, orange, and Peru ground cherry) may

Pulp

serve as potential dietary supplement for the prevention and treatment of cancer because

Seed Antiproliferative activity

of strong antiproliferative activities against the cancer cell lines.  2013 Elsevier Ltd. All rights reserved.

Cancer

1.

Introduction

Cancer is one of the leading chronic diseases and causes death worldwide, represents a major public health problem in the world. Major types of cancer include lung, colorectal, breast and liver cancer, which are the most commonly diagnosed cancers in the world (Carvalho et al., 2010; Mou et al., 2011; Noratto et al., 2010; Wang et al., 2006). The increasing incidence of cancer reported over the last few decades led to development of new anticancer drugs, drug combinations, and chemotherapy strategies by methodical and scientific exploration of enormous pool of synthetic, biological, and natural products (Collett et al., 2010; Mukherjee, Basu, Sarkar,

& Ghosh, 2001; Surh, 1998, 2003; Tang et al., in press). Both chemotherapy and radiation therapy played an important role in the treatment of cancer, but with various strong side effects. Therefore, the search for new compounds that could safely and effectively block or reverse cancer development remains a priority. A large number of plant-derived compounds have been identified for prevention and treatment of cancer, such as paclitaxel, vinblastine, camptothecin, resveratrol, melatonin, indole-3-carbinol, sulforaphane, genistein, brassinin, lycopene and astaxanthin (Deng et al., 2012a; Li et al., 2012a; Mou et al., 2011; Surh, 1998, 2003; Wu et al., 2012). In light of the continuing need for effective anticancer agents and the association of fruits and vegetables consumption

* Corresponding authors. Tel.: +86 20 87332391; fax: +86 20 87330446. E-mail addresses: [email protected] (H.-B. Li), [email protected] (X.-R. Xu). 1 These authors equally contributed to this paper. 1756-4646/$ - see front matter  2013 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.jff.2013.04.016

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with reduced cancer risk, edible plants are increasingly being considered as sources of anticancer drugs (Ferguson, Kurowska, Freeman, Chambers, & Koropatnick, 2004; He & Liu, 2007). Cancer chemoprevention with strategies using natural foods is regarded as one of the most visible fields for cancer control (Li et al., 2012b; Zhou et al., 2012). Epidemiological studies suggested that antioxidant supplements might reduce the risk of cancer recurrence and cancer-related mortality (Suhail et al., 2012), and consuming food and beverages rich in polyphenols was associated with a lower incidence of cancers (Naasani et al., 2003). Experimental investigations demonstrated that many naturally occurring agents and plant extracts showed antioxidant and anticancer potential in a variety of bioassay systems and animal models (Chandrasekara & Shahidi, 2011a, 2012; Chen et al., 2011; Cooper et al., 2011; Deng et al., 2012b; Engel, Oppermann, Falodun, & Kragl, 2011; Kandaswami et al., 2005; Pai et al., 2012). Fruits contain many polyphenolics, such as quercetin, kaempferol, phenolic acids, gallic acid, chlorogenic acid, luteolin, ellagic acid, and protocatechuic acid (Fu et al., 2011), which may protect against cancer. Nevertheless, combinations of polyphenols naturally found in fruits and vegetables have been suggested to be most favorable for cancer prevention and their anticarcinogenic effects (Liu, 2003; Stefanska, Salame, Bednarek, & Fabianowska-Majewska, 2012). Therefore, the objective of this study was to compare the antiproliferative capacities of different portions of 61 fruits against four cancer cell lines A549 (human lung cancer cells), MCF-7 (human breast cancer cells), HepG2 (human hepatoma cells) and HT-29 (human colon cancer cells) using the MTT assay, and to supply new information on the anticancer function of selected fruits for nutritionists and the general public.

2.

Materials and methods

2.1.

Chemical reagents and samples

Dimethylsulfoxide (DMSO), 3-(4,5-dimethylthiazole-2yl)-2,5diphenyl tetrazolium bromide (MTT) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM, pH 7.4), fetal bovine serum and trypsin were obtained from Gibco (Invitrogen, New York, NY, USA). All other chemicals and solvents used in this study were of analytical grade. The 61 fruits were purchased from markets in Guangzhou, China.

2.2.

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was redissolved in dimethylsulfoxide to obtain 200 mg/mL stock solution. Finally, the extract was filtered through a 0.22 lm Millipore filter and stored at 4 C until further use.

2.3.

Cell culture

A549 (human lung cancer cell line), and MCF-7 (human breast cancer cell line) were obtained from the No. 1 hospital affiliated to Sun Yat-Sen University (Zhongshan Road 2, Guangzhou, China). HepG2 (human hepatoma cell line) was provided by the School of Public Health, Sun Yat-Sen University. HT-29 (human colon cancer cell line) was obtained from the No. 6 hospital affiliated to Sun Yat-Sen University. The four cell lines were grown in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum supplemented with 1% penicillin–streptomycin. All the cell lines were maintained in a 5% CO2/37 C incubator (Liu, Flynn, Ferguson, Hoagland, & Yu, 2011; Mou et al., 2011; Yang & Liu, 2009).

2.4. Evaluation of antiproliferative effects of samples on cancer cell lines The effects of the samples on the viability of various cancer cell lines were determined by the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. Cancer cells were seeded in 96-well microtiter plates at a density of 1 · 105 cells/mL in 100 lL DMEM complete medium, and allowed 24 h for attachment. Then the culture medium was replaced with 200 lL medium containing 100 mg/mL sample extract. Control wells received the growth medium without sample, and blank wells contained 200 lL of growth medium with no cells, which were incubated for 48 h at 37 C in a humid atmosphere with 5% CO2. After 48 h, the culture medium was removed and washed with PBS for twice, and 10 lL of sterile filtered MTT solution (5 mg/mL) in phosphate-buffered saline (PBS, pH 7.4) was added to each well, producing a final concentration of 0.5 mg/mL of MTT. After a further incubation of 4 h, the untransformed MTT was carefully removed by pipette and the insoluble formazan crystals were dissolved in 150 lL of DMSO per well, then shaken on a table concentrator for 10 min. The absorbance was measured at 490 nm by an ELX800 microplate spectrophotometer (BIOTEC, Germany) (Liu et al., 2011). Antiproliferative activity was measured as percent compared to control wells. All samples were conducted in triplicate.

2.5. Determination of the dose-dependent manner and IC50 value

Sample preparation

The fresh fruits were washed with deionized water and dried in room temperature, and then their peels, pulps and seeds were separated carefully. Each part of fruit was ground into fine particles with a grinder. A precisely weighed amount (3 g) of the ground sample was extracted with 27 mL of a mixture of ethanol–water (50:50, v/v) at room temperature for 24 h in a shaking water bath (Fu et al., 2011). The mixture was centrifuged at 4200g for 30 min, and the supernatant was collected. Ethanol in the crude extract was removed under vacuum using a rotary evaporator at 50 C. The extract

To evaluate the ability of the samples inhibiting the proliferation of the cultured cells, the MTT method was performed to determine the cell viability as described above. Cancer cells were seeded in 96 well microtiter plates at a density of 1 · 105 cells/mL in 100 lL DMEM complete cell culture medium. After 24 h, the culture medium was replaced with 200 lL media containing various concentrations (20, 40, 60, 80 and 100 mg/mL) of the sample extracts which showed the strongest proliferative inhibition activities. Control wells received the growth medium without samples, and blank wells contained 200 lL of growth medium with no cells,

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which were incubated for 48 h at 37 C, 5% CO2 atmosphere. The culture medium was removed and washed with PBS for twice. Ten microliters of sterile filtered MTT solution (5 mg/ mL) in phosphate-buffered saline (PBS, pH 7.4) was added to each well, producing a final concentration of 0.5 mg/mL of MTT. After 4 h, the untransformed MTT was carefully removed by pipette and the insoluble formazan crystals were dissolved in 150 lL of dimethyl sulphoxide (DMSO) per well, and then shaken on a table concentrator for 10 min. The absorbance was measured at 490 nm using an ELX800 microplate spectrophotometer (BIO-TEC, Germany) (Liu et al., 2011). The data were presented as percentage of variation with respect controls (designed as 100%). The percentage of cell proliferative inhibition over control was calculated by the formula: Inhibition rate ð%Þ ¼

ðcontrol  experimentÞ  100 control

All samples were conducted in triplicate. The IC50 value was calculated as the concentration compared to the culture medium at which cell growth was inhibited by 50% compared to untreated controls.

2.6.

HPLC analysis

The phenolic compounds in samples were analyzed by HPLCPAD according to the method reported in the literature (Deng et al., 2012b; Fu et al., 2011; Sakakibara, Honda, Nakagawa, Ashida, & Kanazawa, 2003). In detail, the HPLC system employed a Waters (Milford, MA, USA) 1525 binary HPLC pump separation module with an auto-injector and a Waters 2996 photodiode array detector (PAD). Separation was performed with an Agilent Zorbax Extend-C18 column (250 mm · 4.6 mm, 5 lm) at 35 C with a gradient elution solution A, composed of acetic acid–water solution (0.1% acetic acid) and methanol (9:1; v/v), and solution B, comprising methanol and acetic acid–water solution (0.1% acetic acid) (7:3; v/v), which delivered at a flow rate of 1.0 mL/min as follows: 0 min, 100% (A); 15 min, 70% (A); 45 min, 65% (A); 65 min, 60% (A); 70 min, 50% (A); and 95 min, 0% (A). The UV spectra were recorded between 190 and 600 nm for peak characterization. Phenolic compounds were quantified by the peak area of maximum absorption wavelength, respectively.

2.7.

Statistical analysis

The results were expressed as mean ± SD (standard deviation). Statistical analysis was conducted using SPSS 17.0. Differences among treatments were determined using ANOVAs. The P value less than 0.05 was considered to be statistically significant.

3.

Results and discussion

3.1. lines

Antiproliferative activities of samples on cancer cell

Lung, breast, liver and colon cancers are among the most frequent cancer types. Cultured cancer cells are valuable reagents for rapid screening of potential anticancer agents

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as well as for elucidation of mechanism of their activities. Therefore, in the present study, the selected fruits that are very common consumed worldwide were evaluated for their abilities to inhibit the growth of human lung (A549), breast (MCF-7), liver (HepG2) and colon (HT-29) cancer cell lines by the MTT assay. MTT can be converted to an insoluble purple formazan by a mitochondrial enzyme in living cells and the amount of formazan produced is directly proportional to the number of viable cells (Barrios et al., 2010; Valente, de Pinho, Henrique, Pereira, & Carvalho, 2012). The in vitro proliferative inhibition of the 162 samples of the peels, pulps and seeds from 61 fruits toward the growth of the four cancer cell lines was shown in Table 1. As seen from Table 1, there was a considerable difference in the sensitivity of these cancer cell lines to the samples (p < 0.05). Totally, Peru ground cherry peel and pulp, hami melon seed, Jiangxi orange pulp, prune seed, and blueness jujube seed showed strong inhibitory effects against all the four cancer cell lines. In details, Peru ground cherry peel, hami melon seed, honey peach seed, green husk orange peel, Jiangxi orange pulp, honey peach pulp, honey peach peel, prune seed, and pineapple peel showed the strongest inhibitory effect toward the growth of A549. Peru ground cherry pulp, blueness jujube peel, blueness jujube seed, little chicken melon seed, hami melon seed, prune seed, crown pear seed, salak seed, and black beauty melon seed showed the strongest inhibitory effect toward the growth of MCF-7. Peru ground cherry pulp and peel, hylocereus undatus peel, honey peach pulp, hawthorn seed, crown pear seed, blueness jujube seed, prune seed, and salak pulp showed the strongest inhibitory effect toward the growth of HepG2. Peru ground cherry pulp, Jiangxi orange pulp, hawthorn seed, orange peel (South Africa), orange pulp (South Africa), pineapple peel, red guava peel, kumquat peel, and navel orange pulp (China) showed the strongest inhibitory effect toward the growth of HT-29. The results indicated that different fruits and different parts of one fruit had diverse capacities against different target cancer cell lines and the variation was very large, and also strongly suggested that the peels, pulps and seeds of some fruits could be promising anticancer agents. It hints that different fruits and different parts of one fruit should be used in different way for the prevention and treatment of different cancers, and fruits could be an important dietary source to prevent cancers.

3.2. Determination of the dose-dependent manner and IC50 value Five different concentrations (20, 40, 60, 80, and 100 mg/mL) of the extracts were applied to inhibit the four cancer cell lines proliferation. Fig. 1 presented the concentration effectiveness of the 10 samples that possessed the strongest activities on viability of A549, MCF-7, HepG2 and HT-29 cancer cell lines. All the test samples exhibited a dose-dependent inhibition against the cell viability and proliferation. Treatment of A549 cells with Peru ground cherry peel produced the greatest dose-dependent response, from 65% to 100% cell growth inhibition at the concentrations ranging from 20 to 100 mg/mL, followed by Jiangxi orange pulp (8–99%), and prune seed (0– 97%) (Fig. 1a). For MCF-7 cells, treatment with these samples resulted in levels of cell growth inhibition ranging from 0%

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Table 1 – Antiproliferative capacities (·100%) of 162 extract samples from 61 fruits on 4 cancer cell lines. No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64

Fruit name

A549

MCF-7

HepG2

HT-29

Prune peel Prune pulp Prune seed Black plum peel Black plum pulp Black plum seed Winter jujube peel Winter jujube pulp Winter jujube seed Import orange peel Import orange pulp Import orange seed Grapefruit peel Grapefruit pulp Grapefruit seed Kiwifruit peel Kiwifruit pulp Pitaya peel Pitaya pulp Crown pear peel Crown pear pulp Crown pear seed Honey peach peel Honey peach pulp Honey peach seed Green husk orange peel Green husk orange pulp Apple peel (rose red) Apple pulp (rose red) Apple seed (rose red) Apple peel (green, imported) Apple pulp (green, imported) Apple seed (green, imported) Apple peel (red, delicious) Apple pulp (red, delicious) Apple seed (red, delicious) Huang jin shuai apple peel Huang jin shuai apple pulp Huang jin shuai apple seed Apple red gala peel Apple red gala pulp Apple red gala seed Shangdong fushi apple peel Shangdong fushi apple pulp Shangdong fushi apple seed Rambutan peel Rambutan pulp Rambutan seed Pear (fragrant) peel Pear (fragrant) pulp Pear (fragrant) seed Red grapefruit peel Red grapefruit pulp Carambola peel Carambola pulp Carambola seed Banana peel Banana pulp Lingwu long jujube peel Lingwu long jujube pulp Lingwu long jujube seed Passiflora edulis peel Passiflora edulis pulp Passiflora edulis seed

0.90 ± 0.02 0.89 ± 0.01 1.00 ± 0.00 0.11 ± 0.04 0.91 ± 0.01 0.97 ± 0.01 0.68 ± 0.03 0.96 ± 0.01 0.97 ± 0.00 0.96 ± 0.00 0.97 ± 0.00 0.99 ± 0.00 0.99 ± 0.00 1.00 ± 0.00 0.95 ± 0.01 0.72 ± 0.02 0.76 ± 0.01 0.95 ± 0.01 0.99 ± 0.00 0.99 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 0.99 ± 0.00 0.99 ± 0.00 0.99 ± 0.00 0.79 ± 0.01 0.87 ± 0.05 0.83 ± 0.00 0.85 ± 0.02 0.90 ± 0.00 0.98 ± 0.00 0.89 ± 0.01 0.86 ± 0.02 1.00 ± 0.00 0.93 ± 0.01 0.99 ± 0.01 0.97 ± 0.01 0.93 ± 0.01 0.97 ± 0.01 0.99 ± 0.00 0.96 ± 0.01 0.99 ± 0.00 0.98 ± 0.00 0.88 ± 0.04 0.98 ± 0.00 0.98 ± 0.01 0.99 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.00 0.99 ± 0.00 0.71 ± 0.05 0.94 ± 0.02 0.91 ± 0.03 0.69 ± 0.04 0.94 ± 0.02 0.86 ± 0.02 0.98 ± 0.00 0.89 ± 0.01 0.99 ± 0.00 0.98 ± 0.00 0.99 ± 0.00

0.90 ± 0.00 0.91 ± 0.00 0.99 ± 0.01 0.00 ± 0.00 0.94 ± 0.00 0.93 ± 0.02 0.70 ± 0.03 0.87 ± 0.01 0.92 ± 0.01 0.97 ± 0.01 0.98 ± 0.01 0.85 ± 0.06 0.87 ± 0.00 0.85 ± 0.08 0.76 ± 0.03 0.73 ± 0.05 0.79 ± 0.04 0.76 ± 0.02 0.84 ± 0.04 0.81 ± 0.03 0.86 ± 0.04 1.00 ± 0.00 0.85 ± 0.04 0.88 ± 0.02 0.97 ± 0.01 0.99 ± 0.00 0.99 ± 0.01 0.79 ± 0.01 0.94 ± 0.00 0.80 ± 0.02 0.80 ± 0.00 0.89 ± 0.00 0.95 ± 0.01 0.81 ± 0.02 0.91 ± 0.00 0.87 ± 0.01 0.96 ± 0.01 0.90 ± 0.03 0.73 ± 0.03 0.81 ± 0.02 0.96 ± 0.02 0.81 ± 0.06 0.88 ± 0.02 0.90 ± 0.01 0.85 ± 0.01 0.77 ± 0.06 0.94 ± 0.01 0.97 ± 0.01 0.92 ± 0.01 0.87 ± 0.02 0.98 ± 0.01 0.83 ± 0.00 0.98 ± 0.00 0.49 ± 0.08 0.82 ± 0.04 0.82 ± 0.00 0.78 ± 0.04 0.77 ± 0.10 0.86 ± 0.02 0.91 ± 0.01 0.93 ± 0.02 0.88 ± 0.01 0.98 ± 0.01 0.98 ± 0.01

0.93 ± 0.00 0.92 ± 0.01 1.00 ± 0.00 0.70 ± 0.08 0.96 ± 0.01 0.91 ± 0.01 0.74 ± 0.07 0.93 ± 0.01 0.97 ± 0.00 0.95 ± 0.01 0.98 ± 0.00 0.99 ± 0.00 1.00 ± 0.00 0.99 ± 0.00 0.88 ± 0.02 0.76 ± 0.01 0.81 ± 0.02 0.94 ± 0.01 0.97 ± 0.01 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.02 0.98 ± 0.01 0.99 ± 0.00 0.95 ± 0.06 0.99 ± 0.00 0.99 ± 0.00 0.83 ± 0.02 0.96 ± 0.00 0.94 ± 0.02 0.87 ± 0.03 0.93 ± 0.01 0.98 ± 0.01 0.98 ± 0.01 0.76 ± 0.05 0.94 ± 0.00 0.93 ± 0.02 0.97 ± 0.00 0.78 ± 0.02 0.76 ± 0.04 0.93 ± 0.00 0.94 ± 0.01 0.90 ± 0.01 0.91 ± 0.02 0.79 ± 0.01 0.77 ± 0.02 0.94 ± 0.00 0.94 ± 0.00 0.96 ± 0.02 0.97 ± 0.02 0.98 ± 0.00 0.94 ± 0.05 0.96 ± 0.01 0.56 ± 0.03 0.77 ± 0.01 0.81 ± 0.05 0.89 ± 0.01 0.81 ± 0.03 0.76 ± 0.02 0.95 ± 0.01 0.90 ± 0.02 0.97 ± 0.01 0.97 ± 0.00 0.97 ± 0.01

0.83 ± 0.03 0.81 ± 0.02 0.98 ± 0.03 0.39 ± 0.10 0.88 ± 0.02 0.72 ± 0.04 0.40 ± 0.06 0.89 ± 0.03 0.91 ± 0.01 0.94 ± 0.03 0.98 ± 0.01 0.79 ± 0.04 0.95 ± 0.04 0.88 ± 0.04 0.59 ± 0.07 0.35 ± 0.08 0.04 ± 0.07 0.67 ± 0.03 0.96 ± 0.01 0.89 ± 0.04 0.86 ± 0.01 0.98 ± 0.01 0.87 ± 0.06 0.91 ± 0.01 0.97 ± 0.00 0.99 ± 0.02 0.97 ± 0.02 0.90 ± 0.03 0.96 ± 0.00 0.88 ± 0.05 0.92 ± 0.02 0.90 ± 0.01 0.98 ± 0.00 0.91 ± 0.01 0.96 ± 0.01 0.98 ± 0.01 0.97 ± 0.00 0.95 ± 0.02 0.83 ± 0.01 0.92 ± 0.00 0.97 ± 0.01 0.98 ± 0.01 0.71 ± 0.01 0.99 ± 0.01 0.92 ± 0.01 0.89 ± 0.01 0.97 ± 0.00 0.98 ± 0.00 0.97 ± 0.01 0.99 ± 0.00 0.99 ± 0.00 0.93 ± 0.01 0.98 ± 0.00 0.75 ± 0.05 0.93 ± 0.01 0.91 ± 0.01 0.63 ± 0.06 0.84 ± 0.03 0.56 ± 0.12 0.96 ± 0.03 0.87 ± 0.01 0.89 ± 0.05 0.97 ± 0.02 0.97 ± 0.01

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Table 1 – Continued No.

Fruit name

A549

MCF-7

HepG2

65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128

Lemon peel Lemon pulp Lemon seed Yunnan pomegranate peel Yunnan pomegranate pulp Yunnan pomegranate seed Wax apple peel Wax apple pulp Import red plum peel Import red plum pulp Import red plum seed Salak peel Salak pulp Salak seed Red grape peel (USA) Red grape pulp (USA) Red grape seed (USA) Black grape peel (USA) Black grape pulp (USA) Grape peel (jufeng) Grape pulp (jufeng) Grape seed (jufeng) Cherry tomato peel Cherry tomato pulp Yacon peel Yacon pulp Guava peel Guava pulp Guava seed Jiangxi orange peel Jiangxi orange pulp Sweetsop peel Sweetsop pulp Sweetsop seed Heart left persimmon peel Heart left persimmon pulp Beijing persimmon peel Beijing persimmon pulp Sweet crisp persimmon peel Sweet crisp persimmon pulp Mango peel (Australia) Mango pulp (Australia) Mango seed (Australia) Pear peel (royal) Pear pulp (royal) Pear seed (royal) Black beauty watermelon peel Black beauty watermelon pulp Black beauty watermelon seed Little chicken melon peel Little chicken melon pulp Little chicken melon seed Muskmelon peel (yellow pulp) Muskmelon pulp (yellow pulp) Muskmelon seed (yellow pulp) Blueness jujube peel Blueness jujube pulp Blueness jujube seed Kumquat peel Kumquat pulp Kumquat seed Pomelo peel Pomelo pulp Red guava peel

0.99 ± 0.00 0.99 ± 0.00 0.98 ± 0.00 0.37 ± 0.02 0.57 ± 0.08 0.72 ± 0.04 0.76 ± 0.01 1.00 ± 0.00 0.61 ± 0.09 0.82 ± 0.02 0.91 ± 0.09 0.99 ± 0.00 0.99 ± 0.00 1.00 ± 0.00 0.81 ± 0.01 0.98 ± 0.00 0.01 ± 0.09 0.62 ± 0.05 0.91 ± 0.03 0.61 ± 0.03 0.98 ± 0.01 0.49 ± 0.04 0.96 ± 0.03 0.99 ± 0.00 0.93 ± 0.01 0.79 ± 0.01 0.76 ± 0.02 0.94 ± 0.01 0.96 ± 0.01 1.00 ± 0.00 0.99 ± 0.00 0.00 ± 0.00 0.97 ± 0.01 0.96 ± 0.01 0.90 ± 0.02 0.67 ± 0.03 0.79 ± 0.01 0.66 ± 0.02 0.98 ± 0.00 0.92 ± 0.03 0.39 ± 0.06 0.97 ± 0.01 0.59 ± 0.03 0.93 ± 0.01 0.89 ± 0.02 0.99 ± 0.00 0.90 ± 0.02 0.88 ± 0.05 0.97 ± 0.04 0.78 ± 0.06 0.97 ± 0.01 0.98 ± 0.00 0.89 ± 0.02 0.88 ± 0.03 0.89 ± 0.04 0.94 ± 0.01 0.85 ± 0.02 0.98 ± 0.01 0.97 ± 0.01 0.73 ± 0.06 0.88 ± 0.01 0.94 ± 0.01 0.93 ± 0.01 0.98 ± 0.00

1.00 ± 0.00 0.96 ± 0.01 1.00 ± 0.00 0.00 ± 0.02 0.03 ± 0.04 0.58 ± 0.09 0.58 ± 0.05 0.92 ± 0.01 0.29 ± 0.09 0.75 ± 0.04 0.88 ± 0.00 0.82 ± 0.06 0.80 ± 0.04 1.00 ± 0.02 0.52 ± 0.09 0.86 ± 0.03 0.89 ± 0.07 0.12 ± 0.07 0.78 ± 0.05 0.29 ± 0.24 1.00 ± 0.00 0.12 ± 0.05 0.92 ± 0.03 0.99 ± 0.00 0.87 ± 0.01 0.49 ± 0.03 0.51 ± 0.02 0.69 ± 0.04 0.92 ± 0.06 0.93 ± 0.07 0.98 ± 0.01 0.00 ± 0.02 0.88 ± 0.12 0.97 ± 0.02 0.74 ± 0.01 0.15 ± 0.02 0.63 ± 0.08 0.13 ± 0.19 1.00 ± 0.01 0.94 ± 0.04 0.04 ± 0.09 1.00 ± 0.01 0.48 ± 0.03 0.98 ± 0.02 0.75 ± 0.04 0.85 ± 0.02 0.71 ± 0.06 0.73 ± 0.09 1.00 ± 0.01 0.77 ± 0.06 0.75 ± 0.06 0.99 ± 0.02 0.70 ± 0.03 0.70 ± 0.02 0.83 ± 0.05 1.00 ± 0.01 0.95 ± 0.04 1.00 ± 0.03 0.88 ± 0.02 1.00 ± 0.00 0.48 ± 0.04 0.90 ± 0.09 0.89 ± 0.01 0.93 ± 0.07

0.99 ± 0.01 0.96 ± 0.01 0.98 ± 0.01 0.36 ± 0.09 0.64 ± 0.09 0.73 ± 0.04 0.71 ± 0.05 0.96 ± 0.03 0.66 ± 0.04 0.82 ± 0.05 0.95 ± 0.01 0.98 ± 0.00 1.00 ± 0.01 0.99 ± 0.02 0.71 ± 0.01 0.96 ± 0.01 0.00 ± 0.11 0.48 ± 0.04 0.85 ± 0.04 0.63 ± 0.06 0.96 ± 0.01 0.43 ± 0.02 0.98 ± 0.01 0.98 ± 0.01 0.86 ± 0.07 0.81 ± 0.03 0.69 ± 0.00 0.87 ± 0.04 0.94 ± 0.01 0.98 ± 0.00 0.99 ± 0.01 0.15 ± 0.20 0.98 ± 0.00 0.97 ± 0.02 0.93 ± 0.02 0.63 ± 0.03 0.73 ± 0.11 0.63 ± 0.06 0.98 ± 0.01 0.93 ± 0.05 0.17 ± 0.02 0.98 ± 0.01 0.41 ± 0.01 0.92 ± 0.02 0.99 ± 0.00 0.98 ± 0.01 0.90 ± 0.03 0.92 ± 0.01 0.99 ± 0.00 0.78 ± 0.06 0.97 ± 0.01 0.98 ± 0.00 0.78 ± 0.03 0.96 ± 0.01 0.94 ± 0.01 0.94 ± 0.02 0.96 ± 0.01 0.99 ± 0.01 0.92 ± 0.05 0.87 ± 0.02 0.67 ± 0.06 0.93 ± 0.02 0.91 ± 0.03 0.98 ± 0.00

HT-29 1.00 ± 0.02 0.97 ± 0.01 0.94 ± 0.01 0.00 ± 0.06 0.00 ± 0.06 0.41 ± 0.12 0.81 ± 0.02 0.99 ± 0.02 0.00 ± 0.05 0.67 ± 0.06 0.93 ± 0.01 0.96 ± 0.05 1.00 ± 0.01 1.00 ± 0.02 0.47 ± 0.08 0.93 ± 0.02 0.00 ± 0.00 0.00 ± 0.00 0.94 ± 0.01 0.58 ± 0.05 1.00 ± 0.02 0.00 ± 0.07 0.99 ± 0.02 1.00 ± 0.01 0.94 ± 0.01 0.84 ± 0.02 0.81 ± 0.05 0.93 ± 0.01 0.97 ± 0.01 0.94 ± 0.01 0.99 ± 0.02 0.00 ± 0.00 0.94 ± 0.01 0.97 ± 0.01 0.92 ± 0.03 0.24 ± 0.14 0.65 ± 0.07 0.94 ± 0.03 0.97 ± 0.02 0.99 ± 0.01 0.00 ± 0.10 0.97 ± 0.01 0.45 ± 0.05 0.92 ± 0.01 0.92 ± 0.02 0.95 ± 0.03 0.87 ± 0.00 0.88 ± 0.03 0.99 ± 0.01 0.85 ± 0.01 0.94 ± 0.02 0.97 ± 0.01 0.88 ± 0.01 0.86 ± 0.04 0.90 ± 0.03 0.97 ± 0.01 0.88 ± 0.03 0.98 ± 0.02 0.99 ± 0.01 0.93 ± 0.03 0.82 ± 0.07 0.90 ± 0.01 0.91 ± 0.01 0.99 ± 0.01 (continued on next page)

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Table 1 – Continued No.

Fruit name

A549

MCF-7

HepG2

HT-29

129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162

Red guava pulp Red guava seed Olive peel Olive pulp Olive seed Durian peel Durian pulp Durian seed Jackfruit peel Jackfruit pulp Jackfruit seed Mangosteen peel Mangosteen pulp Longan peel Longan pulp Longan seed Plantain peel Plantain pulp Hami melon peel Hami melon pulp Hami melon seed Hylocereus undatus peel Hylocereus undatus pulp Pineapple peel Pineapple pulp Navel orange peel (China) Navel orange pulp (China) Orange peel (South Africa) Orange pulp (South Africa) Hawthorn peel Hawthorn pulp Hawthorn seed Peru ground cherry peel Peru ground cherry pulp

0.93 ± 0.01 0.95 ± 0.03 0.64 ± 0.07 0.60 ± 0.03 0.83 ± 0.01 0.95 ± 0.00 0.82 ± 0.01 0.75 ± 0.03 0.72 ± 0.02 0.57 ± 0.05 0.81 ± 0.03 0.18 ± 0.08 0.84 ± 0.01 0.55 ± 0.10 0.98 ± 0.00 0.48 ± 0.06 0.97 ± 0.02 0.99 ± 0.01 0.96 ± 0.02 1.00 ± 0.00 1.00 ± 0.00 0.97 ± 0.01 0.84 ± 0.01 0.99 ± 0.01 0.99 ± 0.01 0.99 ± 0.01 0.99 ± 0.00 0.67 ± 0.06 0.98 ± 0.00 0.98 ± 0.01 0.68 ± 0.02 0.99 ± 0.01 0.99 ± 0.00 0.99 ± 0.01

0.83 ± 0.02 0.87 ± 0.03 0.46 ± 0.02 0.33 ± 0.04 0.73 ± 0.02 0.91 ± 0.01 0.36 ± 0.06 0.78 ± 0.04 0.55 ± 0.06 0.25 ± 0.04 0.49 ± 0.10 0.15 ± 0.03 0.83 ± 0.00 0.00 ± 0.00 0.94 ± 0.01 0.43 ± 0.09 0.90 ± 0.02 0.96 ± 0.03 0.85 ± 0.02 0.98 ± 0.01 0.99 ± 0.00 0.95 ± 0.00 0.58 ± 0.06 0.99 ± 0.00 0.99 ± 0.01 0.99 ± 0.01 0.95 ± 0.01 0.40 ± 0.14 0.91 ± 0.03 0.99 ± 0.01 0.36 ± 0.14 0.98 ± 0.00 0.98 ± 0.01 1.00 ± 0.01

0.89 ± 0.01 0.93 ± 0.02 0.52 ± 0.04 0.25 ± 0.10 0.88 ± 0.05 0.84 ± 0.02 0.75 ± 0.07 0.82 ± 0.02 0.61 ± 0.03 0.47 ± 0.05 0.66 ± 0.04 0.20 ± 0.10 0.85 ± 0.02 0.54 ± 0.07 0.97 ± 0.01 0.44 ± 0.07 0.91 ± 0.01 0.98 ± 0.00 0.91 ± 0.00 0.99 ± 0.00 0.99 ± 0.01 0.99 ± 0.00 0.90 ± 0.03 1.00 ± 0.10 1.00 ± 0.00 0.98 ± 0.01 0.99 ± 0.01 0.70 ± 0.08 0.95 ± 0.01 1.00 ± 0.00 0.68 ± 0.1 1.00 ± 0.01 1.00 ± 0.00 1.00 ± 0.01

0.92 ± 0.01 0.91 ± 0.01 0.40 ± 0.04 0.45 ± 0.13 0.82 ± 0.06 0.82 ± 0.04 0.96 ± 0.01 0.85 ± 0.03 0.83 ± 0.04 0.79 ± 0.06 0.90 ± 0.02 0.00 ± 0.05 0.79 ± 0.04 0.31 ± 0.03 0.94 ± 0.01 0.37 ± 0.09 0.92 ± 0.02 0.98 ± 0.01 0.95 ± 0.01 1.00 ± 0.01 0.98 ± 0.02 0.96 ± 0.01 0.93 ± 0.01 0.99 ± 0.01 1.00 ± 0.01 0.99 ± 0.01 1.00 ± 0.00 1.00 ± 0.00 1.00 ± 0.01 0.60 ± 0.03 0.61 ± 0.06 0.99 ± 0.01 0.99 ± 0.00 1.00 ± 0.01

to 99% with Peru ground cherry pulp, followed by 13–96% with blueness jujube peel and 2–77% with blueness jujube peel (Fig. 1b). For HepG2 cells, the level of cell growth inhibition observed with Peru ground cherry pulp ranged from 98% to 100%, followed by 58–100% with Peru ground cherry peel, and 3–100% with hawthorn seed at the concentrations ranging from 20 to 100 mg/mL (Fig. 1c). For HT-29 cells, treatment with Peru ground cherry pulp displayed the greatest dosedependent response, from 79% to 100% cell growth inhibition, hawthorn seed (0–97%), and Jiangxi orange pulp (0–97%) followed at the concentrations ranging from 20 to 100 mg/mL (Fig. 1d). The IC50 values obtained for antiproliferative activities of the samples toward the growth of A549, MCF-7, HepG2 and HT-29 cancer cell lines in vitro were presented in Table 2. All samples exhibited different growth inhibition activities toward A549 cells (IC50 values between 15.78 and 62.33 mg/ mL), MCF-7 cells (50.09–141.79 mg/mL), HepG2 cells (7.39– 99.76 mg/mL) and HT-29 cells (11.35–108.35 mg/mL). While Peru ground cherry fruit showed the lowest IC50 values signifying a higher antiproliferative efficiency toward the four cancer cell lines, Peru ground cherry peel with the lowest IC50 value of 15.78 ± 0.03 mg/mL for A549, Peru ground cherry pulp with the lowest IC50 value of 50.09 ± 0.02 mg/mL for MCF-7, Peru ground cherry pulp with the lowest IC50 value of

7.39 ± 0.01 mg/mL for HepG2 and Peru ground cherry pulp with the lowest IC50 value of 11.35 ± 0.01 mg/mL for HT-29. From these results, Peru ground cherry fruit showed the strongest inhibitory ability with the lowest IC50 value among all the samples against the four cancer cell lines, and the inhibitory effect of Peru ground cherry ranged from 66% to 100% for A549 cells, 0–99% for MCF-7 cells, 58–100% for the HepG2 and 79–100% for HT-29 cells. Polyphenols are naturally occurring compounds that widely distributed in fruits, and possess various biological activities such as antioxidant, antiinflammatory, antiviral, antithrombogenic, and anticarcinogenic actions (Nijveldt et al., 2001; Stewart, Mullen, & Crozier, 2005; Surh, 2003). We have already determined the total phenolic contents of most fruits with a positive correlation with the antioxidant capacities, and identified some polyphenolic compounds (Fu et al., 2011). Recent studies indicated that polyphenols are the main phytochemicals with antiproliferative property of higher plants (Mertens-Talcott, Lee, Percival, & Talcott, 2006; Zhang, Seeram, Lee, Feng, & Heber, 2008). In this study, some bioactives and their contents in the samples have been identified and determined according to the method reported in the literature (Deng et al., 2012b; Fu et al., 2011; Sakakibara et al., 2003), and were shown in Table 3. The chemical composition of peels and seeds of fruits can be significantly different from

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Fig. 1 – The dose-dependent curve of antiproliferative effects of the 9 top samples on the human lung cancer cell line: (a) A549, (b) MCF-7, (c) HepG2 and (d) HT-29.

Table 2 – IC50 values (mg/mL) of 9 top samples with strongest antiproliferative effects toward 4 cancer cell lines. No.

A549

MCF-7

HepG2

3 22 23 24 25 26 77 78 95 113 116 120 122 123 128 149 150 152 155 156 157 160 161 162

53.18 ± 0.02

78.46 ± 0.03 94.80 ± 0.04

68.83 ± 0.03 59.27 ± 0.02

51.76 ± 0.02 49.79 ± 0.02 32.33 ± 0.01 41.01 ± 0.01

HT-29

40.45 ± 0.02

80.80 ± 0.01 97.42 ± 0.02 42.68 ± 0.01

63.19 ± 0.02 113.67 ± 0.02 61.43 ± 0.05 50.52 ± 0.04 57.24 ± 0.03

62.18 ± 0.03 103.04 ± 0.02 90.57 ± 0.01

23.60 ± 0.02

71.68 ± 0.03 21.81 ± 0.01

62.33 ± 0.01

15.78 ± 0.03 50.09 ± 0.02

54.18 ± 0.03 17.69 ± 0.01 7.39 ± 0.01

89.85 ± 0.02 108.35 ± 0.02 83.27 ± 0.02 84.18 ± 0.01 71.66 ± 0.02

process, which might act as cancer blocking and suppressing agents, preventing the carcinogenic initiation and inhibiting cancer promotion and progression (Russo, 2007; Surh, 2003). In addition, polyphenols can block cancer either through: (i) to suppress nuclear factor-kB activation; (ii) to suppress the activation of activator protein-1 transcription factor; (iii) to suppress protein kinases; (iv) to suppress mitogen activated protein kinases; (v) to suppress growth-factor receptor-mediated pathways; (vi) cell cycle arrest and induct apoptosis; (vii) antioxidant and anti-inflammatory effects; and (viii) to suppress angiogenesis (Chandrasekara & Shahidi, 2011b; Fresco, Borges, Diniz, & Marques, 2006; Khan, Zubair, Ullah, Ahmad, & Hadi, 2011; Sharif et al., 2011). A specific phytochemical or a class of phytochemicals in the tested samples may be responsible for their antiproliferative activities. Taking the complex mechanisms proposed for their chemopreventive properties into consideration, it is likely that antiproliferative effects attributed to polyphenols may be based on additive, synergistic, or antagonistic interactions of many compounds present in these fruits (Liu, 2003). Corroborating this, the peel, pulp and seed of fruits may be an effective auxiliary to prevent or treat cancers, though it is not known exactly which polyphenols are responsible, and further study on the precise compounds and mechanisms responsible for the anticancer activities is still needed.

11.35 ± 0.01

4.

that of pulps of fruits, such as winter jujube, red delicious apple, carambola, longan and hawthorn (Table 3). Numerous phytochemicals derived from edible plants have been reported to interfere with a specific stage of the carcinogenic

Conclusions

The in vitro antiproliferative activities of 162 extract samples of peels, pulps, and seeds from 61 fruits on four cancer cell lines were evaluated by the MTT assay. The results showed that some peels, pulps and seeds strongly inhibited the proliferation of A549, MCF-7, HepG2 and HT-29 and decreased the

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Table 3 – Main phenolic compounds and their contents in fruits (mean ± SD, mg/100 g). Common name (Latin name)

Peel

Prune (Prunus sp.)

Chlorogenic acid 1.30 ± 0.01

Black plum (Plum sp.)

Quercetin 2.57 ± 0.13

Winter jujube (Elaeagnus macrophylla (Thunb) Cogn)

Cyanidin 3-glucoside 2.81 ± 0.08

Grapefruit (Citrus paradisi Macf.)

Gallic acid 1.45 ± 0.10 Protocatechuic acid 1.64 ± 0.10 Chlorogenic acid 1.94 ± 0.20 Cyanidin 3-glucoside 1.78 ± 0.14 Protocatechuic acid 1.27 ± 0.10 Gallic acid 25.61 ± 0.97 Catechin 5.95 ± 0.20 Epicatechin 14.77 ± 0.40 Tangeretin 7.15 ± 0.44 Chlorogenic acid 19.17 ± 0.42 Epicatechin 4.84 ± 0.21

Kiwifruit (Actinidia chinensis) Crown pear (Pyrus spp.)

Green husk orange (Citrus reticulate) Green, imported Apple (Malus domestic)

Pulp Chlorogenic acid 1.26 ± 0.12 Rutin 1.44 ± 0.10 Quercetin 0.50 ± 0.02 Luteolin 1.33 ± 0.17 Kaempferol 0.59 ± 0.05 Gallic acid 3.23 ± 0.25 Homogentisic acid 6.56 ± 0.48 Gallic acid 2.49 ± 0.25 Tangeretin 3.77 ± 0.17

Chlorogenic acid 3.57 ± 0.25 Quercetin 2.45 ± 0.10

Homogentisic acid 4.24 ± 0.13 Catechin 8.79 ± 0.50 nd

Gallic acid 1.67 ± 0.07

nd

Gallic acid 0.45 ± 0.02 Chlorogenic acid 2.33 ± 0.11

Gallic acid 5.39 ± 0.22

Tangeretin 4.03 ± 0.17 Chlorogenic acid 4.76 ± 0.25 Epicatechin 2.81 ± 0.10 Catechin 1.95 ± 0.15 Gallic acid 5.28 ± 0.20

nd

Red delicious apple (Malus pumila Mill.)

Cyanidin 3-glucoside 1.29 ± 0.06 Chlorogenic acid 1.72 ± 0.04

Huang jin shuai apple (Malus domestica)

Gallic acid 2.55 ± 0.23

Gallic acid 1.02 ± 0.07

Shangdong fuji apple (Malus pumila Mill.)

Chlorogenic acid 27.72 ± 1.05 Ferulic acid 4.68 ± 0.32 Gallic acid 10.43 ± 0.75 Homogentisic acid 22.19 ± 0.92 Cyanidin 3-glucoside 25.56 ± 1.43

Chlorogenic acid 47.75 ± 0.85 Ferulic acid 3.12 ± 0.20 Gallic acid 6.43 ± 0.50 Protocatechuic acid 5.81 ± 0.69

Rambutan (Nephelium lappaceum)

Seed

Chlorogenic acid 4.13 ± 0.30 Epicatechin 2.54 ± 0.10

Protocatechuic acid 2.06 ± 0.09 Forulic acid 270.81 ± 5.73 Tangeretin 1.47 ± 0.09 Gallic acid 2.87 ± 0.11 Catechin 6.83 ± 0.34 Forulic acid 475.96 ± 17.19 nd

Gallic acid 2.84 ± 0.33

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Table 3 – Continued Common name (Latin name) Pear (fragrant) (Pyrus sinkiangensis) Red grapefruit (Citrus paradisi Macf)

Carambola (Carambola sp.)

Banana (Musa paradisiacal)

Lemon (Lemon sp.)

Yunnan pomegranate (Punica granatum Linn)

Peel nd Protocatechuic acid 4.51 ± 0.51 Catechin 19.84 ± 0.82 Chlorogenic acid 4.97 ± 0.62 Gallic acid 16.33 ± 0.88 Catechin 81.29 ± 4.60 Epicatechin 154.97 ± 5.84 Catechin 3.15 ± 0.14 Chlorogenic acid 4.62 ± 0.32 Epicatechin 7.93 ± 0.44 Tangeretin 1.80 ± 0.10 Gallic acid 5.41 ± 0.37 Chlorogenic acid 8.48 ± 0.32

Wax apple (Eugenia javanica Lam)

nd

Red grape, USA (Vitis vinifera Linn)

Gallic acid 2.15 ± 0.15 Epicatechin 11.26 ± 0.41 nd

Cherry tomato (Lycopersivonesculentum) Guava (Psidiumguajava L.)

Sweetsop (Annona squamosa L.)

Heart left persimmon (Diospyros kaki Thunb.)

Mango (Mangifera indica Linn)

Gallic acid 1.24 ± 0.10

Catechin 150.42 ± 4.75 Cyanidin 3-glucoside 22.63 ± 1.59 Epicatechin 197.44 ± 3.85 Gallic acid 19.78 ± 0.54

Catechin 7.97 ± 0.64 Tangeretin 1.96 ± 0.08

Pulp

Seed

Chlorogenic acid 0.52 ± 0.03 Cyanidin 3-glucoside 4.37 ± 0.77 Protocatechuic acid 1.67 ± 0.20

nd

Chlorogenic acid 0.55 ± 0.03

Gallic acid 4.03 ± 0.25

Gallic acid 2.87 ± 0.10

nd

Gallic acid 5.45 ± 0.35 Catechin 6.45 ± 0.35 Gallic acid 1.36 ± 0.09 Homogentisic acid 3.6 ± 0.44

nd

Gallic acid 14.84 ± 0.55 Chlorogenic acid 3.30 ± 0.20 nd

Gallic acid 4.28 ± 0.31 Gallic acid 1.58 ± 0.10 Homogentisic acid 5.99 ± 0.23 Kaempferol 4.83 ± 0.30

Gallic acid 10.93 ± 0.31 Catechin 3.07 ± 0.20 Epicatechin 2.75 ± 0.22 Homogentisic acid 4.51 ± 0.33 Protocatechuic acid 2.79 ± 0.17 Chlorogenic acid 2.35 ± 0.22

nd

Homogentisic acid 5.58 ± 0.61 Cyanidin 3-glucoside 2.06 ± 0.11 Chlorogenic acid 5.1 ± 0.44 nd

nd

nd Gallic acid 2.23 ± 0.15 Homogentisic acid 6.00 ± 0.55 Catechin 30.05 ± 0.96 Kaempferol 125.34 ± 3.75

nd

Homogentisic acid 29.63 ± 2.05 Chlorogenic acid 16.88 ± 1.14 Coumaric acid 39.65 ± 1.98 Tangeretin 37.67 ± 1.25 (continued on next page)

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Table 3 – Continued Common name (Latin name)

Peel

Pulp

Royal pear (Pyrus communis Linn.)

Gallic acid 4.79 ± 0.53 Catechin 8.24 ± 0.42

Gallic acid 2.39 ± 0.20 Homogentisic acid 1.52 ± 0.06

Little chicken melon (Diplocyclos Palmatus (L.) C. Jeffr)

Protocatechuic acid 117.68 ± 5.32 Chlorogenic acid 620.38 ± 11.56 Gallic acid 2.49 ± 0.09 Homogentisic acid 3.15 ± 0.10 Tangeretin 1.91 ± 0.10 nd

Gallic acid 2.99 ± 0.33

Kumquat (Fortunella margarita (Lour.) Swingle)

Pomelo (Citrus Paradisi)

Olive (Olea europaea)

Durian (Durio zibethinus Murr.) Jackfruit (Artocarpus heterophyllus) Mangosteen (Garcinia mangostana)

Longan (Dimocarpus longgana Lour.)

Plantain (Musa basjoo) Hami melon (Cucumis melo var. saccharinus) Pineapple (Ananas comosus)

Hawthorn (Crataegus pinnatifida)

Homogentisic acid 116.51 ± 3.31 Epicatechin 19.89 ± 1.10 Chlorogenic acid 17.45 ± 0.55 nd

Homogentisic acid 8.73 ± 0.46 Epicatechin 11.51 ± 1.02 Catechin 7.5 ± 0.45 Chlorogenic acid 15.32 ± 0.71

Gallic acid 5.98 ± 0.44 Catechin 8.25 ± 0.38

Chlorogenic acid 1.22 ± 0.06 Quercetin 0.30 ± 0.04 Homogentisic acid 223.61 ± 6.87 Chlorogenic acid 6.89 ± 0.33

nd

Gallic acid 3.62 ± 0.20

Gallic acid 14.38 ± 0.65 nd Gallic acid 1.83 ± 0.12 Homogentisic acid 2.68 ± 0.09 Catechin 53.12 ± 3.77 Cyanidin 3-glucoside 5.44 ± 0.30 Epicatechin 199.75 ± 6.45

Gallic acid 4.52 ± 0.09 Homogentisic acid 7.35 ± 0.66 Catechin 6.91 ± 0.81 nd

Gallic acid 1.09 ± 0.05

Kaempferol 2.76 ± 0.25 Quercetin 1.47 ± 0.13 nd

nd

Seed

Gallic acid 2.14 ± 0.10 Gallic acid 2.53 ± 0.12 Epicatechin 1.92 ± 0.04

Epicatechin 47.3 ± 1.35

Gallic acid 1.2 ± 0.10

nd nd nd

Homogentisic acid 3.65 ± 0.27 Cyanidin 3-glucoside 3.44 ± 0.15 Chlorogenic acid 17.71 ± 0.99 nd Chlorogenic acid 2.59 ± 0.09 nd

Gallic acid 9.61 ± 0.51 Kaempferol 133.87 ± 4.30

nd: not detected.

viability of these cancer cell lines in a dose-dependent manner. The results also indicated that different samples possess selective antiproliferative effects against different target cancer cell lines. It hints that different fruits and different part of one fruit should be utilized for the prevention and treatment of different cancers in different ways. This study supplied

new information on the antiproliferative function of different part of fruits for nutritionists and the general public. The fruits may be an effective auxiliary to prevent or treat cancers, and further study on the precise compounds and mechanisms responsible for the anticancer activities of these fruits is still needed.

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Acknowledgements This research was supported by the Hundred-Talents Scheme of Chinese Academy of Sciences, and the HundredTalents Scheme of Sun Yat-Sen University.

R E F E R E N C E S

Barrios, J., Cordero, C. P., Aristizabal, F., Heredia, F. J., Morales, A. L., & Osorio, C. (2010). Chemical analysis and screening as anticancer agent of anthocyanin-rich extract from Uva Caimarona (Pourouma cecropiifolia Mart.) fruit. Journal of Agricultural and Food Chemistry, 58, 2100–2110. Carvalho, M., Silva, B. M., Silva, R., Valentao, P., Andrade, P. B., & Bastos, M. L. (2010). First report on Cydonia oblonga Miller anticancer potential: Differential antiproliferative effect against human kidney and colon cancer cells. Journal of Agricultural and Food Chemistry, 58, 3366–3370. Chandrasekara, A., & Shahidi, F. (2011a). Determination of antioxidant activity in free and hydrolyzed fractions of millet grains and characterization of their phenolic profiles by HPLCDAD-ESI-MSn. Journal of Functional Foods, 3, 144–158. Chandrasekara, A., & Shahidi, F. (2011b). Antiproliferative potential and DNA scission inhibitory activity of phenolics from whole millet grains. Journal of Functional Foods, 3, 159–170. Chandrasekara, A., & Shahidi, F. (2012). Bioaccessibility and antioxidant potential of millet grain phenolics as affected by simulated in vitro digestion and microbial fermentation. Journal of Functional Foods, 4, 226–237. Chen, D., Wan, S. B., Yang, H. J., Yuan, J., Chan, T. H., & Dou, Q. P. (2011). EGCG, green tea polyphenols and their synthetic analogs and prodrugs for human cancer prevention and treatment. Advances in Clinical Chemistry, 53, 155–177. Collett, N. P., Amin, A. R. M. R., Bayraktar, S., Pezzuto, J. M., Shin, D. M., Khuri, F. R., Aggarwal, B. B., Surh, Y. J., & Kucuk, O. (2010). Cancer prevention with natural compounds. Seminars in Oncology, 37, 258–281. Cooper, A. J. L., Krasnikov, B. F., Niatsetskaya, Z. V., Pinto, J. T., Callery, P. S., Villar, M. T., Artigues, A., & Bruschi, S. A. (2011). Cysteine S-conjugate beta-lyases: important roles in the metabolism of naturally occurring sulfur and seleniumcontaining compounds, xenobiotics and anticancer agents. Amino Acids, 41, 7–27. Deng, G. F., Xu, X. R., Li, S., Li, F., Xia, E. Q., & Li, H. B. (2012a). Natural sources and bioactivities of resveratrol. International Journal of Modern Biology and Medicine, 1, 1–20. Deng, G. F., Xu, X. R., Guo, Y. J., Xia, E. Q., Li, S., Wu, S., Chen, F., Ling, W. H., & Li, H. B. (2012b). Determination of antioxidant property and their lipophilic and hydrophilic phenolic contents in cereal grains. Journal of Functional Foods, 4, 906–914. Engel, N., Oppermann, C., Falodun, A., & Kragl, U. (2011). Proliferative effects of five traditional Nigerian medicinal plant extracts on human breast and bone cancer cell lines. Journal of Ethnopharmacology, 137, 1003–1010. Ferguson, P. J., Kurowska, E., Freeman, D. J., Chambers, A. F., & Koropatnick, D. J. (2004). A flavonoid fraction from cranberry extract inhibits proliferation of human tumor cell lines. Journal of Nutrition, 134, 1529–1535. Fresco, P., Borges, F., Diniz, C., & Marques, M. P. (2006). New insights on the anticancer properties of dietary polyphenols. Medicinal Research Reviews, 26, 747–766. Fu, L., Xu, B. T., Xu, X. R., Gan, R. Y., Zhang, Y., Xia, E. Q., & Li, H. B. (2011). Antioxidant capacities and total phenolic contents of 62 fruits. Food Chemistry, 129, 345–350.

5 ( 2 0 1 3 ) 1 2 9 8 –1 3 0 9

He, X. J., & Liu, R. H. (2007). Triterpenoids isolated from apple peels have potent antiproliferative activity and may be partially responsible for apple’s anticancer activity. Journal of Agricultural and Food Chemistry, 55, 4366–4370. Kandaswami, C., Lee, L. T., Lee, P. P. H., Hwang, J. J., Ke, F. C., Huang, Y. T., & Lee, M. T. (2005). The antitumor activities of flavonoids. In Vivo, 19, 895–909. Khan, H. Y., Zubair, H., Ullah, M. F., Ahmad, A., & Hadi, S. M. (2011). Oral administration of copper to rats leads to increased lymphocyte cellular DNA degradation by dietary polyphenols: Implications for a cancer preventive mechanism. Biometals, 24, 1169–1178. Li, A. N., Xu, X. R., Li, S., Guo, Y. J., Liu, J. L., & Li, H. B. (2012a). Secretion and bioactivity of melatonin. International Journal of Modern Biology and Medicine, 1, 21–39. Li, F., Xu, X. R., Li, S., Deng, G. F., Wu, S., & Li, H. B. (2012b). Resources and bioactivities of lycopene. International Journal of Food Nutrition and Safety, 1, 15–31. Liu, R. H. (2003). Health benefits of fruit and vegetables are from additive and synergistic combinations of phytochemicals. American Journal of Clinical Nutrition, 78, 517–520. Liu, Y. T., Flynn, T. J., Ferguson, M. S., Hoagland, E. M., & Yu, L. L. (2011). Effects of dietary phenolics and botanical extracts on hepatotoxicity-related endpoints in human and rat hepatoma cells and statistical models for prediction of hepatotoxicity. Food and Chemical Toxicology, 49, 1820–1827. Mertens-Talcott, S. U., Lee, J. H., Percival, S. S., & Talcott, S. T. (2006). Induction of cell death in Caco-2 human colon carcinoma cells by ellagic acid rich fractions from muscadine grapes (Vitis rotundifolia). Journal of Agricultural and Food Chemistry, 54, 5336–5343. Mou, H. B., Zheng, Y., Zhao, P., Bao, H. Y., Fang, W. J., & Xu, N. (2011). Celastrol induces apoptosis in non-small-cell lung cancer A549 cells through activation of mitochondria- and Fas/FasL-mediated pathways. Toxicology In Vitro, 25, 1027–1032. Mukherjee, A. K., Basu, S., Sarkar, N., & Ghosh, A. C. (2001). Advances in cancer therapy with plant based natural products. Current Medicinal Chemistry, 8, 1467–1486. Naasani, I., Oh-hashi, F., Oh-hara, T., Feng, W. Y., Johnston, J., Chan, K., & Tsuruo, T. (2003). Blocking telomerase by dietary polyphenols is a major mechanism for limiting the growth of human cancer cells in vitro and in vivo. Cancer Research, 63, 824–830. Nijveldt, R. J., van Nood, E., van Hoorn, D. E. C., Boelens, P. G., van Norren, K., & van Leeuwen, P. A. M. (2001). Flavonoids: a review of probable mechanisms of action and potential applications. American Journal of Clinical Nutrition, 74, 418–425. Noratto, G. D., Bertoldi, M. C., Krenek, K., Talcott, S. T., Stringheta, P. C., & Mertens-Talcott, S. U. (2010). Anticarcinogenic effects of polyphenolics from mango (Mangifera indica) varieties. Journal of Agricultural and Food Chemistry, 58, 4104–4112. Pai, K. S. R., Srilatha, P., Suryakant, K., Setty, M. M., Nayak, P. G., Rao, C. M., & Baliga, M. S. (2012). Anticancer activity of Berberis aristata in Ehrlich ascites carcinoma-bearing mice: A preliminary study. Pharmaceutical Biology, 50, 270–277. Russo, G. L. (2007). Ins and outs of dietary phytochemicals in cancer chemoprevention. Biochemical Pharmacology, 74, 533–544. Sakakibara, H., Honda, Y., Nakagawa, S., Ashida, H., & Kanazawa, K. (2003). Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, 51, 571–581. Sharif, T., Auger, C., Bronner, C., Alhosin, M., Klein, T., EtienneSelloum, N., Schini-Kerth, V. B., & Fuhrmann, G. (2011). Selective proapoptotic activity of polyphenols from red wine on teratocarcinoma cell, a model of cancer stem-like cell. Investigational New Drugs, 29, 239–247.

JOURNAL OF FUNCTIONAL FOODS

Stefanska, B., Salame, P., Bednarek, A., & Fabianowska-Majewska, K. (2012). Comparative effects of retinoic acid, vitamin D and resveratrol alone and in combination with adenosine analogues on methylation and expression of phosphatase and tensin homologue tumour suppressor gene in breast cancer cells. British Journal of Nutrition, 107, 781–790. Stewart, A. J., Mullen, W., & Crozier, A. (2005). On-line highperformance liquid chromatography analysis of the antioxidant activity of phenolic compounds in green and black tea. Molecular Nutrition & Food Research, 49, 52–60. Suhail, N., Bilal, N., Khan, H. Y., Hasan, S., Sharma, S., Khan, F., Mansoor, T., & Banu, N. (2012). Effect of vitamins C and E on antioxidant status of breast-cancer patients undergoing chemotherapy. Journal of Clinical Pharmacy Therapeutics, 37, 22–26. Surh, Y. J. (1998). Cancer chemoprevention by dietary phytochemicals: A mechanistic viewpoint. Cancer Journal, 11, 6–10. Surh, Y. J. (2003). Cancer chemoprevention with dietary phytochemicals. Nature Reviews Cancer, 3, 768–780. Tang, W. M., Chan, E., Kwok, C. Y., Lee, Y. K., Wu, J. H., Wan, C. W., Chan, R. Y., Yu, P. H., & Chan, S. W. (2012). A review of the anticancer and immunomodulatory effects of Lycium barbarum fruit. Inflammopharmacology, 20, 307–314.

5 ( 2 01 3 ) 12 9 8–13 0 9

1309

Valente, M. J., de Pinho, P. G., Henrique, R., Pereira, J. A., & Carvalho, M. (2012). Further insights into chemical characterization through GC–MS and evaluation for anticancer potential of Dracaena draco leaf and fruit extracts. Food and Chemical Toxicology, 50, 3847–3852. Wang, X. J., Yuan, S. L., Wang, J., Lin, P., Liu, G. J., Lu, Y. R., Zhang, J., Wang, W. D., & Wei, Y. Q. (2006). Anticancer activity of litchi fruit pericarp extract against human breast cancer in vitro and in vivo. Toxicology and Applied Pharmacology, 215, 168–178. Wu, S., Li, S., Xu, X. R., Deng, G. F., Li, F., Zhou, J., & Li, H. B. (2012). Sources and bioactivities of astaxanthin. International Journal of Modern Biology and Medicine, 1, 96–107. Yang, J., & Liu, R. H. (2009). Synergistic effect of apple extracts and quercetin 3-b-d-glucoside combination on antiproliferative activity in MCF-7 human breast cancer cells in vitro. Journal of Agricultural and Food Chemistry, 57, 8581–8586. Zhang, Y., Seeram, N. P., Lee, R., Feng, L., & Heber, D. (2008). Isolation and identification of strawberry phenolics with antioxidant and human cancer cell antiproliferative properties. Journal of Agricultural and Food Chemistry, 56, 670–675. Zhou, J., Li, S., Wu, S., Xu, X. R., Deng, G. F., Li, F., & Li, H. B. (2012). Analytical methods, natural sources and bioactivities of lutein. International Journal of Food Nutrition and Safety, 1, 75–98.