Effects of phenolic compounds of colored wheats on colorectal cancer cell lines

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Öykü Gönül Geyik
Zeynep Hazal Tekin-Cakmak
Vladamir P. Shamanin
Salih Karasu
Inna V. Pototskaya
Sergey S. Shepelev
Alexandr S. Chursin
Alexey I. Morgounov
Mustafa Yaman
Osman Sagdic
Hamit Koksel


blue wheat bran, colorectal cancer cells, free and bound phenolic, HCT-116, HT-29, purple wheat bran, red wheat bran


In this study, the different colored wheat brans were analyzed and compared for phenolic content (PC), phenolic compositions, and the total antioxidant capacity (TEAC) with methods based on the ability to eliminate radicals of 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH), and anthocyanin compositions. This study also aims to characterize the bioactive components of wheat grain genotypes as well as to test the protective and rescuer effects of their extracts on colorectal cancer (CRC) cell lines. PCs in the bound insoluble fraction of red wheat bran, blue wheat bran, and purple wheat bran were determined as 369.60, 446.95, and 486.79 mg gallic acid equivalents (GAE)/100 g wheat bran, respectively, while strong relationships were detected between PC and antioxidant activity (DPPH and ABTS) results. HPLC analysis of phenolic extracts demonstrated that ferulic acid was determined as the dominant phenolic acid in the bound fractions of red, purple, and blue wheats. In the free fractions, p-coumaric acid (11.55 µg/100 g wheat bran) was the dominant phenolic acid for red wheat bran, whereas ellagic acid (14.72 and 11.55 µg/100 g wheat bran) was the highest phenolic acid for purple and blue wheat brans, respectively. In bound fractions, ferulic acid was the highest phenolic acid for red (988.39 µg/100 g wheat bran), purple (1948.76 µg/100 g wheat bran), and blue (2263.96 µg/100 g wheat bran) wheat brans. On the other hand, Cyanidin-3-O-glucoside chloride was the predominant anthocyanin in free extracts of purple and blue wheat brans. In line with the antioxidant activities and phenolic acid concentrations, the blue wheat bran extracts increased CRC cell viability nonsignificantly in HT-29 and HCT-116 cell lines, whereas purple wheat bran extract had a significantly higher (P = 0.0361) rescuer effect compared to vehicle control under 50 µM H2O2 concentration. In conclusion, the in vitro data here show that blue and purple wheat brans are posing a novel means to increase the defense of cells against oxidative stress and cell death.

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Abdel-Aal, E.-S.M., Abou-Arab, A.A., Gamel, T.H., Hucl, P., Young, J.C. and Rabalski, I., 2008. Fractionation of blue wheat anthocyanin compounds and their contribution to antioxidant properties. Journal of Agricultural and Food Chemistry 56(23): 11171–11177. 10.1021/jf802168c

Abdel-Aal, E.-S.M. and Hucl, P., 2003. Composition and stability of anthocyanins in blue-grained wheat. Journal of Agricultural and Food Chemistry 51(8): 2174–2180. 10.1021/jf011210d

Abdel-Aal, E.-S.M., Young, J.C. and Rabalski, I., 2006. Anthocyanin composition in black, blue, pink, purple, and red cereal grains. Journal of Agricultural and Food Chemistry 54(13): 4696–4704. 10.1021/jf0606609

Adam, A., Leuillet, M., Crespy, V., Levrat-Verny, M.-A., Leenhardt, F., Demigné, C. and Rémésy, C., 2002. The bioavailability of ferulic acid is governed primarily by the food matrix rather than its metabolism in intestine and liver in rats. The Journal of Nutrition 132(7): 1962–1968. 10.1093/jn/132.7.1962

Akkoc, Y., Lyubenova, L., Grausgruber, H., Janovská, D., Yazici, A., Cakmak, I. and Gozuacik, D., 2019. Minor cereals exhibit superior antioxidant effects on human epithelial cells compared to common wheat cultivars. Journal of Cereal Science 85: 143–152. 10.1016/j.jcs.2018.12.006

Alazzouni, A.S., Dkhil, M.A., Gadelmawla, M.H., Gabri, M.S., Farag, A.H. and Hassan, B.N., 2021. Ferulic acid as anticarcinogenic agent against 1, 2-dimethylhydrazine induced colon cancer in rats. Journal of King Saud University-Science 33(2): 101354. 10.1016/j.jksus.2021.101354

Arnao, M.B., Cano, A., Alcolea, J.F. and Acosta, M., 2001. Estimation of free radical-quenching activity of leaf pigment extracts. Phytochemical Analysis: An International Journal of Plant Chemical and Biochemical Techniques 12(2): 138–143. 10.1002/pca.571

Buescher, M.I. and Gallaher, D.D., 2014. Wheat color (class), not refining, influences colon cancer risk in rats. Nutrition and Cancer 66(5): 849–856. 10.1080/01635581.2014.904909

Chen, W., Muüller, D., Richling, E. and Wink, M., 2013. Anthocyanin-rich purple wheat prolongs the life span of Caenorhabditis elegans probably by activating the DAF-16/FOXO transcription factor. Journal of Agricultural and Food Chemistry 61(12): 3047–3053. 10.1021/jf3054643

Cheynier, V., Gómez, C. and Ageorges, A., 2012. Flavonoids: anthocyanins. In: Nollet, L.M.C. and Toldrá, F. (eds.) Handbook of analysis of active compounds in functional foods. 1st ed., CRC Press, pp. 379–403.

Cione, E., Tucci, P., Senatore, V., Perri, M., Trombino, S., Iemma, F., Picci, N. and Genchi, G., 2008. Synthesized esters of ferulic acid induce release of cytochrome c from rat testes mitochondria. Journal of Bioenergetics and Biomembranes 40: 19–26. 10.1007/s10863-007-9097-7

Demirci, M., Tomas, M., Tekin-Cakmak, Z.H. and Karasu, S., 2021. Berberis crataegina DC. as a novel natural food colorant source: ultrasound-assisted extraction optimization using response surface methodology and thermal stability studies. Food Science and Technology 42., e03421. 10.1590/fst.13421

El-Gogary, R.I., Nasr, M., Rahsed, L.A. and Hamzawy, M.A., 2022. Ferulic acid nanocapsules as a promising treatment modality for colorectal cancer: preparation and in vitro/in vivo appraisal. Life Sciences 298: 120500. 10.1016/j.lfs.2022.120500

Eroğlu, C., Seçme, M., Bağcı, G. and Dodurga, Y., 2015. Assessment of the anticancer mechanism of ferulic acid via cell cycle and apoptotic pathways in human prostate cancer cell lines. Tumor Biology 36: 9437–9446. 10.1007/s13277-015-3689-3

Garg, M., Chawla, M., Chunduri, V., Kumar, R., Sharma, S., Sharma, N.K., Kaur, N., Kumar, A., Mundey, J.K., Saini, M.K. and Singh, S.P., 2016. Transfer of grain colors to elite wheat cultivars and their characterization. Journal of Cereal Science 71: 138–144. 10.1016/j.jcs.2016.08.004

Gordeeva, E., Badaeva, E., Yudina, R., Shchukina, L., Shoeva, O. and Khlestkina, E., 2019. Marker-assisted development of a blue-grained substitution line carrying the Thinopyrum ponticum chromosome 4Th(4D) in the spring bread wheat Saratovskaya 29 background. Agronomy 9(11): 723. 10.3390/agronomy9110723

Hosseinian, F.S., Li, W. and Beta, T., 2008. Measurement of anthocyanins and other phytochemicals in purple wheat. Food Chemistry 109(4): 916–924. 10.1016/j.foodchem.2007.12.083

Hou, Y., Yang, J., Zhao, G. and Yuan, Y., 2004. Ferulic acid inhibits endothelial cell proliferation through NO down-regulating ERK1/2 pathway. Journal of Cellular Biochemistry 93(6): 1203–1209. 10.1002/jcb.20281

Hudson, E.A., Dinh, P.A., Kokubun, T., Simmonds, M.S. and Gescher, A., 2000. Characterization of potentially chemopreventive phenols in extracts of brown rice that inhibit the growth of human breast and colon cancer cells. Cancer Epidemiology Biomarkers & Prevention 9(11): 1163–1170. Available at: https://pubmed.ncbi.nlm.nih.gov/11097223/

Kawabata, K., Yamamoto, T., Hara, A., Shimizu, M., Yamada, Y., Matsunaga, K., Tanaka, T. and Mori, H., 2000. Modifying effects of ferulic acid on azoxymethane-induced colon carcinogenesis in F344 rats. Cancer Letters 157(1): 15–21. 10.1016/S0304-3835(00)00461-4

Lachman, J., Martinek, P., Kotíková, Z., Orsák, M. and Šulc, M., 2017. Genetics and chemistry of pigments in wheat grain–a review. Journal of Cereal Science 74: 145–154. 10.1016/j.jcs.2017.02.007

Li, W., Shan, F., Sun, S., Corke, H. and Beta, T., 2005. Free radical scavenging properties and phenolic content of Chinese black-grained wheat. Journal of Agricultural and Food Chemistry 53(22), 8533–8536. 10.1021/jf051634y

Liu, Q., Qiu, Y. and Beta, T., 2010. Comparison of antioxidant activities of different colored wheat grains and analysis of phenolic compounds. Journal of Agricultural and Food Chemistry 58(16): 9235–9241. 10.1021/jf101700s

Liyana-Pathirana, C.M. and Shahidi, F., 2006. Importance of insoluble--bound phenolics to antioxidant properties of wheat. Journal of Agricultural and Food Chemistry 54(4): 1256–1264. 10.1021/jf052556h

Liyana-Pathirana, C.M. and Shahidi, F., 2007. The antioxidant potential of milling fractions from breadwheat and durum. Journal of Cereal Science 45(3): 238–247. 10.1016/j.jcs.2006.08.007

Ma, D., Li, Y., Zhang, J., Wang, C., Qin, H., Ding, H., Xie, Y. and Guo, T., 2016. Accumulation of phenolic compounds and expression profiles of phenolic acid biosynthesis-related genes in developing grains of white, purple, and red wheat. Frontiers in Plant Science 7: 528. 10.3389/fpls.2016.00528

Mazewski, C. and Gonzalez de Mejia, E., 2018. Impact of anthocyanins on colorectal cancer. In: Advances in plant phenolics: from chemistry to human health. American Chemical Society, pp. 339–370. 10.1021/bk-2018-1286.ch019

Mazewski, C., Liang, K. and de Mejia, E.G., 2017. Comparison of the inhibitory potential of anthocyanin-rich plant extracts on colon cancer cell proliferation and their mechanism of action in vitro. The FASEB Journal 31(S1): 300–307. 10.1096/fasebj.31.1_supplement.300.7

Mazewski, C., Liang, K. and Gonzalez de Mejia, E., 2018. Comparison of the effect of chemical composition of anthocyanin--rich plant extracts on colon cancer cell proliferation and their potential mechanism of action using in vitro, in silico, and biochemical assays. Food Chemistry 242: 378–388. 10.1016/j.foodchem.2017.09.086

Mazewski, C.E., 2019. Inhibitory potential of anthocyanins and anthocyanin-rich extracts on colorectal cancer progression through apoptosis, angiogenesis, and immune response effects. Available at: https://hdl.handle.net/2142/105188

Ransy, C., Vaz, C., Lombès, A. and Bouillaud, F., 2020. Use of H(2)O(2) to cause oxidative stress, the catalase issue. International Journal of Molecular Sciences 21(23), 9149. 10.3390/ijms21239149

Rice-Evans, C.A., Miller, N.J. and Paganga, G., 1996. Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Radical Biology and Medicine 20(7): 933–956. 10.1016/0891-5849(95)02227-9

Roy, N., Narayanankutty, A., Nazeem, P., Valsalan, R., Babu, T. and Mathew, D., 2016. Plant phenolics ferulic acid and p-coumaric acid inhibit colorectal cancer cell proliferation through EGFR down-regulation. Asian Pacific Journal of Cancer Prevention 17(8): 4019–4023. 10.14456/apjcp.2016.208/APJCP.2016.17.8.4019

Saini, P., Kumar, N., Kumar, S., Mwaurah, P.W., Panghal, A., Attkan, A.K., Singh, V.K., Garg, M.K. and Singh, V., 2021. Bioactive compounds, nutritional benefits and food applications of colored wheat: a comprehensive review. Critical Reviews in Food Science and Nutrition 61(19): 3197–3210. 10.1080/10408398.2020.1793727

Serpen, A., Goükmen, V., Karagoüz, A. and Koüksel, H., 2008. Phytochemical quantification and total antioxidant capacities of emmer (Triticum dicoccon Schrank) and einkorn (Triticum monococcum L.) wheat landraces. Journal of Agricultural and Food Chemistry 56(16): 7285–7292. 10.1021/jf8010855

Shamanin, V.P., Tekin-Cakmak, Z.H., Gordeeva, E.I., Karasu, S., Pototskaya, I., Chursin, A.S., Pozherukova, V.E., Ozulku, G., Morgounov, A.I., Sagdic, O. and Koksel, H., 2022. Antioxidant capacity and profiles of phenolic acids in various genotypes of purple wheat. Foods 11(16): 2515. 10.3390/foods11162515

Sharma, S., Chunduri, V., Kumar, A., Kumar, R., Khare, P., Kondepudi, K.K., Bishnoi, M. and Garg, M., 2018. Anthocyanin biofortified colored wheat: nutritional and functional characterization. PLoS One 13(4): e0194367. 10.1371/journal.pone.0194367

Sharma, A., Shahzad, B., Rehman, A., Bhardwaj, R., Landi, M. and Zheng, B., 2019. Response of phenylpropanoid pathway and the role of polyphenols in plants under abiotic stress. Molecules 24(13): 2452. 10.3390/molecules24132452

Sharma, N., Tiwari, V., Vats, S., Kumari, A., Chunduri, V., Kaur, S., Kapoor, P. and Garg, M., 2020. Evaluation of anthocyanin content, antioxidant potential and antimicrobial activity of black, purple and blue colored wheat flour and wheat-grass juice against common human pathogens. Molecules 25(24): 5785. 10.3390/molecules25245785

Siebenhandl, S., Grausgruber, H., Pellegrini, N., Del Rio, D., Fogliano, V., Pernice, R. and Berghofer, E., 2007. Phytochemical profile of main antioxidants in different fractions of purple and blue wheat, and black barley. Journal of Agricultural and Food Chemistry 55(21): 8541–8547. 10.1021/jf072021j

Singh, R., Chidambara Murthy, K. and Jayaprakasha, G., 2002. Studies on the antioxidant activity of pomegranate (Punica granatum) peel and seed extracts using in vitro models. Journal of Agricultural and Food Chemistry 50(1): 81–86. 10.1021/jf010865b

Singh Tuli, H., Kumar, A., Ramniwas, S., Coudhary, R., Aggarwal, D., Kumar, M., Sharma, U., Chaturvedi Parashar, N., Haque, S., and Sak, K., 2022. Ferulic acid: a natural phenol that inhibits neoplastic events through modulation of oncogenic signaling. Molecules 27(21): 7653. 10.3390/molecules27217653

Singleton, V.L. and Rossi, J.A., 1965. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture 16(3): 144–158. 10.5344/ajev.1965.16.3.144

Toda, S., Kumura, M. and Ohnishi, M., 1991. Effects of phenolcarboxylic acids on superoxide anion and lipid peroxidation induced by superoxide anion. Planta Medica 57(1): 8–10. 10.1055/s-2006-960005

Tyl, C.E. and Bunzel, M., 2012. Antioxidant activity-guided fractionation of blue wheat (UC66049 Triticum aestivum L.). Journal of Agricultural and Food Chemistry 60(3): 731–739. 10.1021/jf203648x

Vichai, V. and Kirtikara, K., 2006. Sulforhodamine B colorimetric assay for cytotoxicity screening. Nature Protocols 1(3): 1112–1116. 10.21769/BioProtoc.1984

Wink, M., 2008. Evolutionary advantage and molecular modes of action of multi-component mixtures used in phytomedicine. Current Drug Metabolism 9(10): 996–1009. 10.2174/138920008786927794

Zheng, Y., You, X., Guan, S., Huang, J., Wang, L., Zhang, J. and Wu, J., 2019. Poly (ferulic acid) with an anticancer effect as a drug nanocarrier for enhanced colon cancer therapy. Advanced Functional Materials 29(15): 1808646. 10.1002/adfm.201808646

Žilić, S., 2016. Phenolic compounds of wheat. Their content, antioxidant capacity and bioaccessibility. MOJ Food Processing & Technology 2(3): 037. 10.15406/mojfpt.2016.02.00037

Žilić, S., Basić, Z., Hadži-Tašković Šukalović, V., Maksimović, V., Janković, M. and Filipović, M., 2014. Can the sprouting process applied to wheat improve the contents of vitamins and phenolic compounds and antioxidant capacity of the flour? International Journal of Food Science & Technology 49(4): 1040–1047. 10.1111/ijfs.12397

Žilić, S., Serpen, A., Akıllıoğlu, G., Janković, M. and Gökmen, V., 2012. Distributions of phenolic compounds, yellow pigments and oxidative enzymes in wheat grains and their relation to antioxidant capacity of bran and debranned flour. Journal of Cereal Science 56(3): 652–658. 10.1016/j.jcs.2012.07.014

Žilić, S., Šukalović, V.H.-T., Dodig, D., Maksimović, V., Maksimović, M. and Basić, Z., 2011. Antioxidant activity of small grain cereals caused by phenolics and lipid soluble antioxidants. Journal of Cereal Science 54(3): 417–424. 10.1016/j.jcs.2011.08.006