This study aims to elucidate the beneficial effect of Punica granatum L., Lythraceae (pomegranate) peel extract in the management of colon cancer induced intrarectally with N-methylnitrosourea. Adult male Sprague-Dawley rats were administered N-methylnitrosourea (2 mg in 0.5 ml water/rat) intrarectally three times/week for five weeks to induce colorectal cancer, followed by treatment with either 5-fluorouracil (12.5 mg/kg, i.p.) or Punica peel extract (2.25 or 4.5 g/kg, p.o.). Developed tumor elevated plasma TGF-β, and Bcl2, serum epidermal growth factor, carcinoembryonic antigen, colon cancer specific antigens, and matrix metalloproteinase-7. Besides, immune-histochemical studies revealed an increase in COX-2, cyclin D1 and survivin content, as well as upregulation of the expression of colonic β-Catenin, K-ras and C-myc genes. These results were further supported by the histological findings. Punica peel extract-treated rats, particularly those treated with a high dose, exhibited a marked reduction in the aforementioned parameters and improved the histological organization of the colon tissue. These alterations were consistent with those mediated through 5-fluorouracil. The present study encourages the use of P. granatum L. against colon cancer. Because Punica peel extract promotes apoptosis, mitigates inflammation and suppresses tumor cell proliferation in vivo, the potential mechanism underlying these activities might depend on the inhibition of the Wnt/β-Catenin signaling pathway.
Adhami, V.M., Khan, N., Mukhtar, H., 2009. Cancer chemoprevention by pomegranate: laboratory and clinical evidence. Nutr. Cancer 61, 811–815.
Ahmed, H.H., Abdel-Rahman, M., Salem, F.E.H., Shalby, A.B., Lokman, M.S., 2013. Antitumor efficacy of Boswella serrata extract in management of colon cancer induced in experimental animal. Int. J. Pharm. Pharmaceut. Sci. 5, 379–389.
Ashihara, E., Takada, T., Maekawa, T., 2015. Targeting the canonical Wnt/β-Catenin pathway in hematological malignancies. Cancer Sci. 106, 665–671.
Austinat, M., Dunsch, R., Wittekind, C., Tannapfel, A., Gebhardt, R., Gaunitz, F., 2008. Correlation between beta-Catenin mutations and expression of Wnt-signaling target genes in hepatocellular carcinoma. Mol. Cancer, https://doi.org/10.1186/1476-4598-7-21.
Bhatia, D., Thoppil, R.J., Mandai, A., Samtani, K.A., Darvesh, A.S., Bishayee, A., 2013. Pomegranate bioactive constituents suppress cell proliferation and induce apoptosis in an experimental model of hepatocellular carcinoma: role of Wnt/β-Catenin signaling pathway. Evid Based Complement. Alternat. Med., https://doi.org/10.1155/2013/371813.
Chen, H., Miao, Ci, Geng, M., Liu, J., Hu, Y., Tian, L., Pan, J., Yang, Y., 2013. Anti-tumor effect of rutin on human neuroblastoma cell lines through inducing G2/M cell cycle arrest and promoting apoptosis. Sci. World J., https://doi.org/10.1155/2013/269165.
El-Toumy, S.A., Rauwald, H.W., 2002. Two ellagitannins from Punica granatum heart-wood. Phytochemistry 61, 971–974.
Eshak, M.G., Ghaly, I.S., Khalil, W.K.B., Farag, I.M., Ghanem, K.Z., 2010. Genetic alterations induced by toxic effect of thermally oxidized oil and protective role of tomatoes and carrots in mice. J. Am. Sci. 6, 175–188.
Fuentes-Calvo, I., Blázquez-Medela, A.M., Santos, E., López-Novoa, J.M., Martínez-Salgado, C., 2010. Analysis of k-ras nuclear expression in fibroblasts and mesangial cells. PLoS One 14, e8703.
González-Sarrías, A., Espín, J.C., Tomás-Barberán, F.A., García-Conesa, M.T., 2009. Gene expression, cell cycle arrest and MAPK signalling regulation in Caco-2 cells exposed to ellagicacid and its metabolites, urolithins. Mol. Nutr. Food Res. 53, 686–688.
Herbst, A., Jurinovic, V., Krebs, S., Thieme, S.E., Blum, H., Göke, B., Kolligs, F.T., 2014. Comprehensive analysis of β-Catenin target genes in colorectal carcinoma cell lines with deregulated Wnt/β-Catenin signaling. BMC Genomics, https://doi.org/10.1186/1471-2164-15-74.
Kikuchi, M., Mikami, T., Sato, T., Tokuyama, W., Araki, K., Watanabe, M., Saigenji, K., Okayasu, I., 2009. High Ki67, Bax, and thymidylate synthase expression well correlates with response to chemoradiation therapy in locally advanced rectal cancers: proposal of a logistic model for prediction. Br. J. Cancer 101, 116–123.
Kundu, J.K., Choi, ICY., Surh, Y.J., 2006. beta-Catenin-mediated signaling: a novel molecular target for chemoprevention with anti-inflammatory substances. Biochim. Biophys. Acta 1765, 14–24.
Larrosa, M., Yanéz-Gascón, M.J., Selma, M.V., González-Sarrías, A., Toti, S., Cerón, J.J., Tomás-Barberán, F., Dolara, P., Espín, J.C., 2009. Effect of a low dose of dietary resveratrol on colon microbiota, inflammation and tissue damage in a DSS-induced colitis rat model. J. Agric. Food Chem. 57, 2211–2220.
Longley, D.B., Latif, T., Boyer, J., Allen, W.L., Maxwell, P.J., Johnston, P.G., 2003. The interaction of thymidylate synthase expression with p53-regulated signaling pathways in tumor cells. Semin. Oncol. 30, 3–9.
Middha, S.K., Usha, T., Pande, V., 2013. A review on antihyperglycemic and antihep-atoprotectiveactivity of eco-friendly Punica granatum peel waste. Evid Based Complement. Alternat. Med., https://doi.org/10.1155/2013/656172.
Narisawa, T., Fukaura, Y., 2003. Prevention by intrarectal 5-aminosalicylicacid of N-methylnitrosourea-induced colon cancer in F344 rats. Dis. Colon Rectum 46, 900–903.
Orgil, O., Schwartz, E., Baruch, L., Matityahu, I., Mahajna, J., Amir, R., 2014. The antioxidative and antiproliferative potential of non-edible organs of the pomegranate fruit and tree. LWT-Food Sci. Technol. 58, 571–577.
Patel, R., Ingle, A., Maru, G.B., 2008. Polymeric black tea polyphenols inhibit 1,2-dimethylhydrazine induced colorectal carcinogenesis by inhibiting cell proliferation via Wnt/beta-Catenin pathway. Toxicol. Appl. Pharmacol. 227, 136–146.
Reeves, P.G., Nielsen, F.H., Fahey, G.C., 1993. AIN-93 purified diets for laboratory rodents: final report of the American Institute of 365 Nutrition ad hoc writing committee on the reformulation of the AIN-76A rodent diet. J. Nutr. 123, 1939–1951.
Refaat, B., El-Shemi, A.G., Kensara, O.A., Mohamed, A.M., Idris, S., Ahmad, J., Khojah, A., 2015. Vitamin D3 enhances the tumouricidal effects of 5-fluorouracil through multipathway mechanisms in azoxymethane rat model of colon cancer. J. Exp. Clin. Cancer Res. 34, 71.
Sharma, M., Li, L., Celver, J., Killian, C., Kovoor, A., Seeram, N.P., 2010. Effects of fruit ellagitannin extracts, ellagicacid, and their colonic metabolite, urolithin A, on Wnt signaling. J. Agric. Food Chem. 58, 3965–3969.
Shin, J.W., Seol, I.C., Son, C.G., 2010. Interpretation of animal dose and human equivalent dose for drug development. J. Korean Oriental Med. 31, 1–7.
Shirode, A.B., Kovvuru, P., Chittur, S.V., Henning, S.M., Heber, D., Reliene, R., 2014. Antiproliferative effects of pomegranate extract in MCF-7 breast cancer cells are associated with reduced DNA repair gene expression and induction of double strand breaks. Mol. Carcinog. 53, 458–470.
Srimuangwong, K, Tocharus, C., Tocharus, J., Suksamrarn, A., Chintana, P.Y., 2012. Effects of hexahydrocurcumin in combination with 5-fluorouracil on dimethylhydrazine-induced colon cancer in rats. World J. Gastroenterol. 18, 6951–6959.
Tang, J.M., Min, J., Li, B.S., Hong, S.S., Liu, C., Hu, M., Li, Y., Yang, J., Hong, L., 2016. Therapeutic effects of punicalagin against ovarian carcinoma cells in association with β-Catenin signaling inhibition. Int. J. Gynecol. Cancer 26, 1557–1563.
Tao, L., Kramer, P.M., Wang, W., Yang, S., Lubet, R.A., Steele, V.E., Pereira, M.A., 2002. Altered expression of c-myc, pl6 and p27 in rat colon tumors and its reversal by short-term treatment with chemopreventive agents. Carcinogenesis 23, 1447–1454.
Vijay, M., Sivagami, G., Thayalan, K., Nalini, N., 2016. Radiosensitizing potential of rutin against human colon adenocarcinoma HT-29 cells. Bratisl. Lek. Listy. 117, 171–178.
Watson, S.A., Michael, D., Justin, T.A., Grimes, S., Morris, T.M., Robinson, G., Clarke, P.A., Hardcastle, J.D., 1998. Preclinical evaluation of the gastrimmune immuno-gen alone and in combination with 5-fluorouracil/leucovorin in a rat colorectal cancer model. Int. J. Cancer 75, 873–877.
Wen, W., Ding, J., Sun, W., Wu, K., Ning, B., Gong, W., He, G., Huang, S., Ding, X., Yin, P., Chen, L., Liu, Q., Xie, W., Wang, H., 2010. Suppression of cyclin D1 by hypoxia-inducible factor-1 via direct mechanism inhibits the proliferation and 5-fluorouracil-induced apoptosis of A549 cells. Cancer Res. 70, 2010–2019.
Zhao, Y., Miao, G., Li, Y., Isaji, T., Gu, J., Li, J., Qi, R., 2014. Microrna 130b suppresses migration and invasion of colorectal cancer cells through downregulation of integrin βl. PLoS One 9, e87938.
MBS (PhD student) contributed in collecting and running the laboratory work. HHA supervised the laboratory work. HHA, HSE and NFA contributed in writing the manuscript. HHA and HSE contributed in designing the study, critical analysis of data, supervised the laboratory work. EAKH contributed to molecular and HPLC analysis. All the authors have read the final manuscript and approved the submission.
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The authors declare no conflicts of interest.
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Ahmed, H.H., El-Abhar, H.S., Hassanin, E.A.K. et al. Punica granatum suppresses colon cancer through downregulation of Wnt/β-Catenin in rat model. Rev. Bras. Farmacogn. 27, 627–635 (2017). https://doi.org/10.1016/j.bjp.2017.05.010
- Colon cancer