Abstract
Pomegranate (Punica granatum) is known to contain polyphenols with many potential health benefits, including anti-tumoral, anti-inflammatory, and anti-microbial properties. It has been used in popular medicine for cancer treatment, which still represents the major cause of cancer-related deaths in men worldwide. Importantly, pomegranate peels are valuable by-products of the food industry that are rich in polyphenols. Here we report a comparison between juice and peel aqueous extracts in prostate cancer DU-145 and PC-3 cell lines. Both extracts were able to inhibit the proliferation, migration and colony formation of those cells, although peel extracts presented more robust effects compared to juice. Besides, the growth-related mTOR/S6K signaling pathway presented strong inhibition after pomegranate extracts treatment. This study presents evidence that both juice and isolated peel extracts from promegate fruit have important anti-cancer effects against prostate cancer cells, modulating the mTOR/S6K signaling pathway.
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Introduction
Despite recent advances in research, prostate cancer still represents a major cause of cancer-related morbidity and mortality in men worldwide [1]. The mTOR (mammalian target of rapamycin) signaling pathway is an important controller of cell energy expenditure, signaling to protein synthesis and cell growth and considered an important modulated pathway in cancer [2, 3]. Some natural compounds, when evaluated in early stages of clinical trials, have been shown to be tolerated, safe and effective against several cancer types, generating several efforts in using these compounds for cancer therapy [4]. Moreover, some natural products can reduce adverse effects caused by chemotherapy and radiotherapy, responsible to reduce the quality of life in cancer patients [5].
Pomegranate (Punica granatum) is a member of the Lythraceae family and contains polyphenols with many potential health benefits, such as antioxidant, anti-microbial, anti-inflammatory and anti-cancer properties, including action in the cell cycle, proliferation, invasion and angiogenesis [6]. Pomegranate fruit, including juice, peels, and oil, has been traditionally used in popular medicine against cancer in different cultures and societies for centuries [7, 8]. According to Turrini et al. [9], pomegranate presents relevant beneficial effects, among several other natural compounds used for the prevention and treatment of prostate cancer.
Ellagitannins (ex. punicalagins), punicic acid, flavanoids, anthocyanidins, anthocyanins, and estrogenic flavonols and flavones are therapeutically known significant constituents of pomegranate [10]. Ellagitannins belong to the polyphenol group of hydrolyzable tannins [10]. After their hydrolysis, they liberate ellagic acid and after microbiota assisted processing they also liberate urolithins [11]. Ellagitannins, including ellagic acid, are bioactive polyphenols present in the pomegranate fruit juice that have chemopreventive potential against multiple human carcinomas [12]. The cytotoxic activity has also been related to ursolic acid, ellagic acid, luteolin, and punicic acid from Punica extracts [10].
Several pre-clinical and clinical research studies present evidence of the anti-cancer effects of pomegranate extracts, most using juice-based extracts against cancer cell lines [10, 12]. This report aimed for a comparison between juice and peel aqueous extracts in prostate cancer DU-145 and PC-3 cell lines, assessing proliferation, migration, colony formation of those cells and the Akt/mTOR/S6K signaling pathway status.
Material and Methods
Cell Culture
PC-3 human metastatic prostate cancer cells and DU-145 human metastatic prostate cancer cells were grown as previously described [3].
Pomegranate Extracts Preparation
1 g of lyophilized peels extract or lyophilized pomegranate juice was placed in 10 mL of deionized water. The mixture was homogenized for 1 min on a vortex. The extraction was performed in an ultrasonic bath at 39 °C for 45 min. The extracts were then centrifuged (TDL80-2B) at 5000 rpm for 10 min at 4 °C and subsequently filtered to obtain aqueous extracts. The quantification of total phenolic content was based on a method published by Singleton et al. [13]. The absorbance was measured at 740 nm using a microplate spectrophotometer (Biotek Instruments, Germany). The total phenol content was calculated using a curve prepared with gallic acid (calibration range 20–120 μg/mL) and expressed in terms of gallic acid equivalents per dry weight (mg of GAE/g of DW).
Antioxidant Activity of Pomegranate Peel and Juice Extracts
The antioxidant activity of pomegranate extracts was analyzed by the ferric reducing antioxidant power (FRAP) method, based on the Benzie and Strain methodology [14]. The antioxidant activity was expressed as ferrous sulfate equivalents per dry weight (μmol of Fe+2/g of DW) and calculated using the calibration curve (calibration range 100–700 μM).
Quantification of Phenolics Content by Chromatography
The extracts were analyzed by high-performance liquid chromatography in the EXTRAC-US system (FAPESP 2013/04304–4 – patent pending). Separation of the compound found in the sample was performed with a C18 column (Kinetex C18, 2.6 μm, 100 A, 100 × 4.6 mm, Phenomenex, Torrance, CA, USA). Chromatograms were recorded at 370 nm. These conditions were based on previously developed methods [15]. The identification of the compounds present in the extracts was based on the comparison of the retention time of authentic standards of punicalagin and ellagic acid (Sigma Aldrich, USA). Results were expressed as mg of phenolic compound/L of extract and the concentration of unidentified peaks was as equivalents of other phenolic compound/L of extract.
Proliferation Test
Cultures of PC-3 and DU-145 cells were plated in 24-well plates at 5 × 104 cells / well and incubated with extracts for 24 h or 48 h with 10% FBS at 37 °C. Counts were performed with an automated cell counter (Invitrogen) and Trypan blue exclusion staining.
Scratch Migration Assay
Cultures of PC-3 and DU-145 cells were seeded at a density of 5 × 105 cells/well in 6-well plates and incubated until confluence. Thereafter, the incubated cell cultures were scratched with a p200 pipette tip in the middle of the wells and the culture media were replaced by serum-free media [3]. The scratch area was analyzed by microscope and images were captured at times 0 h, 24 h, and 48 h. The scratch areas were quantified using ImageJ software.
Colonies Formation Assay
Cultures of PC-3 and DU-145 cells were plated at low density (5 × 103 cells/dish) in 60 mm dishes and treated for 10 days, with changes in media containing pomegranate extracts every two days. The cells were washed with PBS, stained with methylene blue (3%) in methanol for 30 min and counted. Colonies smaller than 1 mm in diameter were excluded from counting [3].
Western Blotting
Proteins, in a total of 30 μg per sample, were separated by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and transferred onto nitrocellulose membranes. Western blotting was performed as previously described [3]. The following antibodies and dilutions were used: anti-pS6K1-Thr389; anti-S6K1; anti-pS6-Ser240/244; anti-S6; anti-pAkt-Ser473; anti-Akt; anti-pmTOR-Ser2448; anti-mTOR (Cell Signaling Technologies), diluted 1:1000; and anti-α-tubulin (Calbiochem), diluted 1:5000.
Statistical Analysis
The values presented are means ± standard deviation (SD). The statistical analyses were evaluated by ANOVA tests followed by Bonferroni’s or Dunnet’s post-test, using the software GraphPad Prism 5. The values of p < 0.05 were considered significant. Outliers test.
Results and Discussion
The total phenolic content and the antioxidant activity of the aqueous extracts of the juice and pomegranate peel were quantified by the Folin-Ciocalteau reagent for the polyphenols and by the iron ion reduction power for the antioxidant capacity and results were 390.77 ± 4.64 and 2043.33 ± 101.52 μmol of Fe+2/g of DW for juice and peel extract, respectively, and 23.87 ± 0.34 and 136.27 ± 1.34 mg of AGE/g of DW, for juice and peel extract, respectively. Thus, peel extracts presented higher concentrations of phenolics and FRAP values compared to juice. The extracts were further characterized for the main phenolics composition by liquid chromatography. Chromatograms of the juice and peel extracts are shown in Fig. 1, respectively, and the concentration of compounds present in the extract is presented in Table I.
The main phenolic compounds identified in the samples were α and β-punicalagin, ellagic acid and three of its derivatives (hexoside, pentoside, and deoxihexoside). In the peel extract there was a predominance of punicalagin followed by high amounts of ellagic acid and its derivatives, the pomegranate juice had a higher relative concentration of two very polar unidentified compounds (peaks #1 and #2 - Fig. 1 and Table I). With the exception of these two compounds the concentration of all identified compounds was lower in the pomegranate juice when compared to the peel extract. Only small amounts of punicalagin have been detected in the juice and the concentration of ellagic acid was almost three times lower than in the peel extract. The total concentration of the main detected compounds in the peel extract was almost six times higher than in the pomegranate juice.
Interestingly, in terms of polyphenols, pomegranate peels are one of the most valuable by-products of the food industry [16]. A study has demonstrated that the enrichment of other fruit juices with dried extract of pomegranate peel increased antioxidant activity [17]. About 40–50% of the total pomegranate fruit weight corresponds to peel, which is a significant source of bioactive compounds such as flavonoids, phenolics, ellagitannins, punicalagin and punicalin [16]. Pomegranates peels and leaves are most traditionally used to treat stomach disorders and diarrhea, although several studies link its anti-inflammatory and antioxidant properties to cancer protection [18]. The chromatographic analysis revealed a phenolic profile of the peel extract and juice consistent with the information available in the literature, which reports higher concentration of punicalagin in the peels and mesocarps compared to the juice [19]. The high concentration of punicalagin in the peel extract is particularly relevant since it is a derivative of ellagic acid. Ellagic acid has demonstrated chemopreventive potential against prostate cancer, activating pro-apoptotic genes in an animal model [12]. Several other phenolic compounds, like resveratrol and epigallocatechin gallate, have been reported to have anti-tumorigenic effects against prostate cancer cells [20, 21].
In order to compare the effects of juice and peel extracts in prostate cancer cells, both were diluted at close concentrations and added to cultures of PC-3 and DU-145. After 24 and 48 h, all concentrations presented cytostatic effects in DU-145 cells (Fig. 2A), and only juice at 23.9 μg/mL and peel extract at 27.3 μg/mL did not inhibit PC-3 after 24 h (Fig. 2B). Peel extracts presented a more robust effect in PC-3 after 48 h than juice at similar concentrations.
To evaluate the effects of juice and peel extracts in the migration of prostate cancer cells, a scratch assay has been performed in serum-free conditions. All tested conditions show that juice and peel extracts are able to inhibit the migration of PC-3 and DU-145 cells in monolayer culture (Fig. 3A and B). Representative images of each culture at 0 and 48 h are presented (Fig. 3C), showing a prominent effect on cell migration. Migration is a feature of cancer cells that is deregulated in advanced and metastatic stages of tumor progression and other studies have shown the potency of pomegranate extracts to inhibit migration of lung and ovarian cancer cells through impairment of metalloproteinases [22]. Albrecht et al. [23] studied pomegranate pericarp, fermented juice polyphenols and seed oil extracts on cell cycle, proliferation, apoptosis, gene expression regulation, invasion and in vivo prostate tumor growth [23]. DU-145 cell line presented a significant increase of 11% in G2/M cell cycle phase by treatment with seed oil. For PC-3 cells, all agents suppressed invasion through Matrigel. Pomegranate pericarp polyphenols and seed oil also demonstrated potent inhibition of PC-3 xenograft growth in athymic mice.
The effects of juice and peel extracts on colony formation have also been tested and results are presented in Fig. 4. Both PC-3 and DU-145 presented colony formation inhibition in the presence of pomegranate extracts, whereas peel extracts were more efficient for that inhibition (Fig. 4A and B). Representative images of each culture are presented in Fig. 4C. PC-3 cells present strong metastatic potential and are androgen-independent, and although DU-145 cells are also highly proliferative and androgen-independent, they have moderate metastatic capability [8]. Indeed our results show that PC-3 cells are more resistant to anti-cancer effects of pomegranate extracts compared to DU-145, as seen by proliferation assay and colony formation (Figs. 2 and 4).
In order to explore cell signaling mechanisms related to the effects of pomegranate treatments in prostate cancer cells, we have evaluated the mTOR signaling pathway status after 48 h of treatments. mTOR pathway was evaluated since it is a key controller of cell growth and metabolism and several studies report the deregulation of this pathway in prostate cancer [2, 3, 24]. Pomegranate juice has been diluted at 2.4 μg/mL and peels extract to 13.6 μg/mL. As seen in Fig. 5, significant reductions in phosphorylation levels of Akt, S6K1, and S6 have been observed for DU-145 for the peel extract (Fig. 5A-E), including juice for Akt. For PC-3, inactivation of mTOR has been observed for both juice and peel extract (Fig. 5F-J).
The mTOR pathway can be inhibited directly or indirectly by several natural compounds, including phytochemicals present in pomegranate, and is frequently activated in several human cancers, being considered an attractive therapeutic target for cancer therapy [2, 4]. The consumption of pomegranate juice ad libitum for 10 weeks in rats suppressed the number of crypt foci and inhibited PI3K/Akt phosphorylation and mTOR expression in HT-29 colon cancer cell line [25]. In another study using rats, pomegranate extracts decreased p70S6K and RPS6, as well as Rps6ka2, Map 2 k2, and Mapk1 mRNA [26]. Ellagic acid presented in pomegranate peel extract is able to inhibit the Akt/mTOR signaling pathway by increasing the expression level of IGFBP7 [27]. Finally, inhibition of prostate cancer growth and metastasis due to oral supplementation with pomegranate fruit extract has been reported in transgenic TRAMP (transgenic adenocarcinoma of the mouse prostate) mice, most likely through inhibition of IGF-I/Akt/mTOR [28]. Punicalagin has been reported to inhibit the mTOR signaling pathway in other cell models, in agreement with the results presented in Fig. 5 [29].
As PC-3 has a robust activation of PI3K/Akt due to the PTEN loss, whereas DU-145 does not [30], inhibitory effects of pomegranate extracts in this pathway may be limited, as shown in Fig. 5. Several studies were also able to show different molecular responses between DU-145 and PC-3 [31, 32]. Besides, inhibition of the Akt/mTOR pathway may in part explain the reduction of proliferation, migration and colonies formation in DU145 cells, since this is a central pathway in controlling apoptosis and autophagy [2]. These effects on the mTOR pathway, however, were not robust in PC-3 cells, even with changes in proliferation, migration and colony formation. Therefore, we were able to find molecular differences in the response to pomegranate extracts in those cell lines.
It is well known that diets rich in phytochemicals have been associated with a reduced risk of diseases including several types of cancer, inflammation, neurodegenerative and cardiovascular diseases [19]. Besides, there is an inverse association of cancer incidence/mortality and vegetable and fruit consumption [9]. Phase II clinical trials have associated consumption of pomegranate juice with prolongation of PSA (Prostate-Specific Antigen) doubling time in men diagnosed with prostate carcinoma [9]. Changes in microbiota have also been demonstrated after pomegranate consumption and microbiota-derived metabolites, like urolithins and ellagic acid, were demonstrated to reach the human prostate tissue [33].
Conclusions
In this study we show that juice and isolated peel extracts from pomegranate are able to inhibit proliferation, migration and colony formation of prostate cancer cell lines, regulating the mTOR signaling pathway, a master controller of cell growth and metabolism. Besides, we present strong evidence that aqueous extracts from pomegranate peels, usually a by-product in food processing, have stronger anti-cancer effects when compared to juice from pomegranate. These results may have an impact on the therapeutics of cancer and for food industry related to pomegranate.
Abbreviations
- mTOR:
-
Mammalian Target Of Rapamycin
- FBS:
-
Fetal bovine serum
- PLC:
-
High-Performance Liquid Chromatography
- TRAMP:
-
Transgenic adenocarcinoma of the mouse prostate
References
Bashir MN (2015) Epidemiology of prostate cancer. Asian Pac J Cancer Prev 16:5137–5141
Tavares MR, Pavan ICB, Amaral CL et al (2015) The S6K protein family in health and disease. Life Sci 131:1–10. https://doi.org/10.1016/j.lfs.2015.03.001
Amaral CL, Freitas LB, Tamura RE, Tavares MR, Pavan IC, Bajgelman MC, Simabuco FM (2016) S6Ks isoforms contribute to viability, migration, docetaxel resistance and tumor formation of prostate cancer cells. BMC Cancer 16:602. https://doi.org/10.1186/s12885-016-2629-y
Tan HK, Moad AIH, Tan ML (2014) The mTOR signalling pathway in cancer and the potential mTOR inhibitory activities of natural phytochemicals. Asian Pac J Cancer Prev 15:6463–6475
Zhang Q-Y, Wang F-X, Jia K-K, Kong L-D (2018) Natural product interventions for chemotherapy and radiotherapy-induced side effects. Front Pharmacol 9:1253. https://doi.org/10.3389/fphar.2018.01253
Bassiri-Jahromi S (2018) Punica granatum (pomegranate) activity in health promotion and cancer prevention. Oncol Rev 12:345. https://doi.org/10.4081/oncol.2018.345
Sharma P, McClees SF, Afaq F (2017) Pomegranate for prevention and treatment of cancer: an update. Molecules 22(1):pii:E177. https://doi.org/10.3390/molecules22010177
Wang L, Martins-Green M (2014) Pomegranate and its components as alternative treatment for prostate cancer. Int J Mol Sci 15:14949–14966. https://doi.org/10.3390/ijms150914949
Turrini E, Ferruzzi L, Fimognari C (2015) Potential effects of pomegranate polyphenols in cancer prevention and therapy. Oxidative Med Cell Longev 2015:938475. https://doi.org/10.1155/2015/938475
Vini R, Sreeja S (2015) Punica granatum and its therapeutic implications on breast carcinogenesis: a review. Biofactors 41:78–89. https://doi.org/10.1002/biof.1206
Tomás-Barberán FA, González-Sarrías A, García-Villalba R et al (2017) Urolithins, the rescue of “old” metabolites to understand a “new” concept: metabotypes as a nexus among phenolic metabolism, microbiota dysbiosis, and host health status. Mol Nutr Food Res 61(1). https://doi.org/10.1002/mnfr.201500901
Naiki-Ito A, Chewonarin T, Tang M et al (2015) Ellagic acid, a component of pomegranate fruit juice, suppresses androgen-dependent prostate carcinogenesis via induction of apoptosis. Prostate 75:151–160. https://doi.org/10.1002/pros.22900
Singleton VL, Orthofer R, Lamuela-Raventós RM (1999) [14] analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. 299:152–178
Benzie IF, Strain JJ (1996) The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem 239:70–76. https://doi.org/10.1006/abio.1996.0292
Rostagno MA, Manchón N, D’Arrigo M et al (2011) Fast and simultaneous determination of phenolic compounds and caffeine in teas, mate, instant coffee, soft drink and energetic drink by high-performance liquid chromatography using a fused-core column. Anal Chim Acta 685:204–211. https://doi.org/10.1016/j.aca.2010.11.031
Sreekumar S, Sithul H, Muraleedharan P, Azeez JM, Sreeharshan S (2014) Pomegranate fruit as a rich source of biologically active compounds. Biomed Res Int 2014:686921. https://doi.org/10.1155/2014/686921
Mastrodi Salgado J, Baroni Ferreira TR, de Oliveira BF, Dos Santos Dias CT (2012) Increased antioxidant content in juice enriched with dried extract of pomegranate (Punica granatum) peel. Plant Foods Hum Nutr 67:39–43. https://doi.org/10.1007/s11130-011-0264-y
Khwairakpam AD, Bordoloi D, Thakur KK, Monisha J, Arfuso F, Sethi G, Mishra S, Kumar AP, Kunnumakkara AB (2018) Possible use of Punica granatum (pomegranate) in cancer therapy. Pharmacol Res 133:53–64. https://doi.org/10.1016/j.phrs.2018.04.021
Fischer UA, Carle R, Kammerer DR (2011) Identification and quantification of phenolic compounds from pomegranate (Punica granatum L.) peel, mesocarp, aril and differently produced juices by HPLC-DAD-ESI/MS(n). Food Chem 127:807–821. https://doi.org/10.1016/j.foodchem.2010.12.156
Bettuzzi S, Brausi M, Rizzi F, Castagnetti G, Peracchia G, Corti A (2006) Chemoprevention of human prostate cancer by oral administration of green tea catechins in volunteers with high-grade prostate intraepithelial neoplasia: a preliminary report from a one-year proof-of-principle study. Cancer Res 66:1234–1240. https://doi.org/10.1158/0008-5472.CAN-05-1145
Li G, Rivas P, Bedolla R, Thapa D, Reddick RL, Ghosh R, Kumar AP (2013) Dietary resveratrol prevents development of high-grade prostatic intraepithelial neoplastic lesions: involvement of SIRT1/S6K axis. Cancer Prev Res (Phila) 6:27–39. https://doi.org/10.1158/1940-6207.CAPR-12-0349
Liu H, Zeng Z, Wang S, Li T, Mastriani E, Li QH, Bao HX, Zhou YJ, Wang X, Liu Y, Liu W, Hu S, Gao S, Yu M, Qi Y, Shen Z, Wang H, Gao T, Dong L, Johnston RN, Liu SL (2017) Main components of pomegranate, ellagic acid and luteolin, inhibit metastasis of ovarian cancer by down-regulating MMP2 and MMP9. Cancer Biol Ther 18:990–999. https://doi.org/10.1080/15384047.2017.1394542
Albrecht M, Jiang W, Kumi-Diaka J, Lansky EP, Gommersall LM, Patel A, Mansel RE, Neeman I, Geldof AA, Campbell MJ (2004) Pomegranate extracts potently suppress proliferation, xenograft growth, and invasion of human prostate cancer cells. J Med Food 7:274–283. https://doi.org/10.1089/jmf.2004.7.274
Dubrovska A, Kim S, Salamone RJ, Walker JR, Maira SM, García-Echeverría C, Schultz PG, Reddy VA (2009) The role of PTEN/Akt/PI3K signaling in the maintenance and viability of prostate cancer stem-like cell populations. Proc Natl Acad Sci USA 106:268–273. https://doi.org/10.1073/pnas.0810956106
Banerjee N, Kim H, Talcott S, Mertens-Talcott S (2013) Pomegranate polyphenolics suppressed azoxymethane-induced colorectal aberrant crypt foci and inflammation: possible role of miR-126/VCAM-1 and miR-126/PI3K/AKT/mTOR. Carcinogenesis 34:2814–2822. https://doi.org/10.1093/carcin/bgt295
Kim H, Banerjee N, Ivanov I, Pfent CM, Prudhomme KR, Bisson WH, Dashwood RH, Talcott ST, Mertens-Talcott SU (2016) Comparison of anti-inflammatory mechanisms of mango (Mangifera indica L.) and pomegranate (Punica granatum L.) in a preclinical model of colitis. Mol Nutr Food Res 60:1912–1923. https://doi.org/10.1002/mnfr.201501008
Guo H, Zhang D, Fu Q (2016) Inhibition of cervical cancer by promoting IGFBP7 expression using ellagic acid from pomegranate peel. Med Sci Monit 22:4881–4886
Adhami VM, Siddiqui IA, Syed DN, Lall RK, Mukhtar H (2012) Oral infusion of pomegranate fruit extract inhibits prostate carcinogenesis in the TRAMP model. Carcinogenesis 33:644–651. https://doi.org/10.1093/carcin/bgr308
Wang Y, Chen B, Longtine MS, Nelson DM (2016) Punicalagin promotes autophagy to protect primary human syncytiotrophoblasts from apoptosis. Reproduction 151:97–104. https://doi.org/10.1530/REP-15-0287
Cunningham D, You Z (2015) In vitro and in vivo model systems used in prostate cancer research. J Biol Methods 2(1):pii:e17. https://doi.org/10.14440/jbm.2015.63
Litvinov IV, Antony L, Dalrymple SL, Becker R, Cheng L, Isaacs JT (2006) PC3, but not DU145, human prostate cancer cells retain the coregulators required for tumor suppressor ability of androgen receptor. Prostate 66:1329–1338. https://doi.org/10.1002/pros.20483
Jayakumar S, Kunwar A, Sandur SK, Pandey BN, Chaubey RC (2014) Differential response of DU145 and PC3 prostate cancer cells to ionizing radiation: role of reactive oxygen species, GSH and Nrf2 in radiosensitivity. Biochim Biophys Acta 1840:485–494. https://doi.org/10.1016/j.bbagen.2013.10.006
González-Sarrías A, Giménez-Bastida JA, García-Conesa MT et al (2010) Occurrence of urolithins, gut microbiota ellagic acid metabolites and proliferation markers expression response in the human prostate gland upon consumption of walnuts and pomegranate juice. Mol Nutr Food Res 54:311–322. https://doi.org/10.1002/mnfr.200900152
Acknowledgements
This research has been funded by The São Paulo Research Foundation (grant numbers: FMS, 2012/13558-7 and 2018/14818-9; AECA, 2015/07299-7; RMNB, 2016/06457-0; MAR, 2013/04304-4; fellowship numbers: ICBP, 2015/003111; LBF, 2013/13002-1) and from CNPq (FMS, 447553/2014-3; MAR, 303568/2016-0).
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FMC and LGS have performed cell culture experiments. ICBP has performed western blotting analysis. MAR has performed phenolics analysis. AECA, RMNB, and FMS have designed experiments and written the manuscript. The authors declare no conflict of interest.
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Chaves, F.M., Pavan, I.C.B., da Silva, L.G.S. et al. Pomegranate Juice and Peel Extracts are Able to Inhibit Proliferation, Migration and Colony Formation of Prostate Cancer Cell Lines and Modulate the Akt/mTOR/S6K Signaling Pathway. Plant Foods Hum Nutr 75, 54–62 (2020). https://doi.org/10.1007/s11130-019-00776-0
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DOI: https://doi.org/10.1007/s11130-019-00776-0