Secondary Metabolites in Flax Root Extracts at Various Stages of Maturity and Effects on Proliferation and Cytotoxicity in Oestrogen-Receptor-Positive Breast Cancer Cells
Affiliation(s)
Department of Obstetrics and Gynaecology, Faculty of Medicine, University of Rostock, Rostock, Germany. Institute for Land-Use, Faculty of Agricultural and Environmental Sciences, University of Rostock, Rostock, Germany.
Department of Obstetrics and Gynaecology, Faculty of Medicine, University of Rostock, Rostock, Germany. Institute for Land-Use, Faculty of Agricultural and Environmental Sciences, University of Rostock, Rostock, Germany.
ABSTRACT
Flax contains large amounts of hormone-like compounds, especially lignans. These socalled phytoestrogens are thought to inhibit the cell growth of hormone-sensitive cancers. Hence, we analysed the influence of flax root extracts at various stages of maturity on the proliferation and cytotoxicity in oestrogen-receptor-positive breast cancer cells (MCF7) in vitro. Flax root extracts were prepared by using lignan extraction. The extracted compounds were analysed by Pyrolysis-Field Ionisation Mass Spectrometry. Various extract concentrations were applied to the cells to test for proliferation (BrdU test) and cytotoxicity (LDH test). A significantly higher inhibition of cell proliferation was observed with an extract made from 9-week-old flax roots in comparison with that of 3- and 6-week-old roots. Older roots contained more lignans and other phenolic substances than younger roots. The maturity grade of plants or their various parts is thus important for the production and concentration of secondary metabolites and leads to different biological effects on breast cancer cell growth.
Flax contains large amounts of hormone-like compounds, especially lignans. These socalled phytoestrogens are thought to inhibit the cell growth of hormone-sensitive cancers. Hence, we analysed the influence of flax root extracts at various stages of maturity on the proliferation and cytotoxicity in oestrogen-receptor-positive breast cancer cells (MCF7) in vitro. Flax root extracts were prepared by using lignan extraction. The extracted compounds were analysed by Pyrolysis-Field Ionisation Mass Spectrometry. Various extract concentrations were applied to the cells to test for proliferation (BrdU test) and cytotoxicity (LDH test). A significantly higher inhibition of cell proliferation was observed with an extract made from 9-week-old flax roots in comparison with that of 3- and 6-week-old roots. Older roots contained more lignans and other phenolic substances than younger roots. The maturity grade of plants or their various parts is thus important for the production and concentration of secondary metabolites and leads to different biological effects on breast cancer cell growth.
Cite this paper
Szewczyk, M. , Strater, N. , Schlichting, A. , Briese, V. and Richter, D. (2014) Secondary Metabolites in Flax Root Extracts at Various Stages of Maturity and Effects on Proliferation and Cytotoxicity in Oestrogen-Receptor-Positive Breast Cancer Cells. Advances in Biological Chemistry, 4, 91-97. doi: 10.4236/abc.2014.42013.
Szewczyk, M. , Strater, N. , Schlichting, A. , Briese, V. and Richter, D. (2014) Secondary Metabolites in Flax Root Extracts at Various Stages of Maturity and Effects on Proliferation and Cytotoxicity in Oestrogen-Receptor-Positive Breast Cancer Cells. Advances in Biological Chemistry, 4, 91-97. doi: 10.4236/abc.2014.42013.
References
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http://dx.doi.org/10.3109/07853899709113696 [3] Horn-Ross, P.L., Barnes, S., Lee, M., Coward, L., Mandel, J.E., Koo, J., John, E.M. and Smith, M. (2000) Assessing Phytoestrogen Exposure in Epidemiologic Studies: Development of a Database (United States). Cancer Causes & Control, 11, 289-298.
http://dx.doi.org/10.1023/A:1008995606699 [4] Dambroth, M. and Seehuber, R. (1988) Flachs—Züchtung, Anbau, Verarbeitung. Ulmer, Stuttgart. [5] Diepenbrock, W., Fischbeck, G., Heyland, K.U. and Knauer, N. (1999) Spezieller Pflanzenanbau. Ulmer, Stuttgart. [6] Düll, R. and Kutzelnigg, H. (2005) Taschenlexikon der Pflanzen Deutschlands. Quelle & Meyer, Wiebelsheim. [7] Abarzua, S., Szewczyk, M., Gailus, S., Richter, D.-U., Ruth, W., Briese, V. and Piechulla, B. (2007) Effects of Phytoestrogen Extracts from Linum usitatissimum on the Jeg3 Human Trophoblast Tumour Cell Line. Anticancer Research, 27, 2053-2058. [8] Luyengi, L., Suh, N., Fong, H., Pezzuto, J. and Konghorn, D. (1996) A Lignan and Four Terpenoids from Brucea javanica That Induce Differentiation with Cultured HL-60 Promyelotic Leukemia Cells. Phytochemistry, 43, 409-412.
http://dx.doi.org/10.1016/0031-9422(96)00258-0 [9] Matscheski, A., Richter, D.-U., Hartmann, A.M., Effmert, U., Jeschke, U., Kupka, M.S., Abarzua, S., Briese, V., Ruth, W., Kragl, U. and Piechulla, B. (2006) Effects of Phytoestrogen Extracts Isolated from Rye, Green and Yellow Pea Seeds on Hormone Production and Proliferation of Trophoblast Tumor Cells Jeg3. Hormone Research, 65, 276-288.
http://dx.doi.org/10.1159/000092591 [10] Schulten, H.R. and Halket, J.M. (1986) Rapid Characterisation of Biomaterials by Field Ionization. Organic Mass Spectrometry, 21, 613-622.
http://dx.doi.org/10.1002/oms.1210211004 [11] Schulten, H., Simmleit, N. and Mueller, R. (1989) Characterization of Plant Materials by Pyrolysis-Field Ionization Mass Spectrometry: High-Resolution Mass Spectrometry, Time-Resolved High-Resolution Mass Spectrometry, and Curie-Point Pyrolysis-Gas Chromatography/Mass Spectrometry of Spruce Needles. Analytical Chemistry, 61, 221-227.
http://dx.doi.org/10.1021/ac00178a007 [12] Hempfling, R., Simmleit, N. and Schulten, H.R. (1991) Characterization and Chemodynamics of Plant Constituents during Maturation, Senescence and Humus Genesis in Spruce Ecosystems. Biogeochemistry, 13, 27-60.
http://dx.doi.org/10.1007/BF00002875 [13] Hempfling, R. and Schulten, H.R. (1991) Pyrolysis-(Gas Chromatography/) Mass Spectrometry of Agricultural Soils and Their Humic Fractions. Zeitschrift für Pflanzenernährung und Bodenkunde, 154, 425-430.
http://dx.doi.org/10.1002/jpln.19911540606 [14] Booth, N.L., Overk, C.R., Yao, P., Totura, S., Deng, Y., Hedayat, A.S., Bolton, J.L., Pauli, G.F. and Farnsworth, N.R. (2006) Seasonal Variation of Red Clover (Trifolium pratense, L. fabaceae) Isoflavones and Estrogenic Activity. Journal of Agricultural and Food Chemistry, 54, 1277-1282.
http://dx.doi.org/10.1021/jf052927u [15] Godin, B., Lamaudière, S., Agneessens, R., Schmit, T., Goffart, J.P., Stilmant, D., Gerin, P.A. and Delcarte, J. (2013) Chemical Characteristics and Biofuels Potentials of Various Plant Biomasses: Influence of the Harvesting Date. Journal of the Science of Food and Agriculture, 93, 3216-3224.
http://dx.doi.org/10.1002/jsfa.6159 [16] Abarzua, S., Serikawa, T., Szewczyk, M., Richter, D.U., Piechulla, B. and Briese, V. (2012) Antiproliferative Activity of Lignans against the Breast Carcinoma Cell Lines MCF 7 and BT 20. Archives of Gynecology and Obstetrics, 285, 1145-1151.
http://dx.doi.org/10.1007/s00404-011-2120-6 [17] Theil, C., Briese, V., Gerber, B. and Richter, D.U. (2011) The Effects of Different Lignans and Isoflavones, Tested as Aglycones and Glycosides, on Hormone Receptor-Positive and -Negative Breast Carcinoma Cells in Vitro. Archives of Gynecology and Obstetrics, 284, 459-465.
http://dx.doi.org/10.1007/s00404-010-1661-4 [18] Liu, X., Suzuki, N., Santosh Laxmi, Y.R., Okamoto, Y. and Shibutani, S. (2012) Anti-Breast Cancer Potential of Daidzein in Rodents. Life Sciences, 91, 415-419.
http://dx.doi.org/10.1016/j.lfs.2012.08.022 [19] Ono, M., Koga, T., Ueo, H. and Nakano, S. (2012) Effects of Dietary Genistein on Hormone-Dependent Rat Mammary Carcinogenesis Induced by Ethyl Methanesulphonate. Nutrition and Cancer, 64, 1204-1210.
http://dx.doi.org/10.1080/01635581.2012.718035 [20] Lin, C.C., Tsai, Y.L., Ho, C.T. and Teng, S.C. (2008) Determination of the Differential Estrogenicity of Isoflavonoids by E2-ER-ERE Dependent Gene Expression in Recombinant Yeast and MCF-7 Human Breast Cancer Cells. Food Chemistry, 108, 719-726.
http://dx.doi.org/10.1016/j.foodchem.2007.11.020 [21] Mousavi, Y. and Adlercreutz, H. (1992) Enterolactones and Estradiol Inhibit Each Other’s Proliferative Effect on MCF7 Breast Cancer Cells in Culture. The Journal of Steroid Biochemistry and Molecular Biology, 41, 615-619.
http://dx.doi.org/10.1016/0960-0760(92)90393-W [22] Wang, C. and Kurzer, M.S. (1997) Phytoestrogen Concentration Determines Effects on DNA Synthesis in Human Breast Cancer Cells. Nutrition and Cancer, 28, 236-247.
http://dx.doi.org/10.1080/01635589709514582 [23] Welshons, W.V., Murphy, C.S., Koch, R., Calaf, G. and Jordan, V.C. (1987) Stimulation of Breast Cancer Cells in Vitro by the Environmental Estrogen Enterolactone and the Phytoestrogen Equol. Breast Cancer Research and Treatment, 10, 169-175.
http://dx.doi.org/10.1007/BF01810580
[1] Kuiper, G.G., Carlsson, B., Grandien, K., Enmark, E., Häggblad, J., Nilsson, S., Nilsson, S. and Gustafsson, J.A. (1997) Comparison of the Ligand Binding Specificity and Transcript Tissue Distribution of Estrogen Receptors Alpha and Beta. Endocrinology, 138, 863-870. [2] Adlercreutz, H. and Mazur, W. (1997) Phyto-Oestrogens and Western Diseases. Annals of Medicine, 29, 95-120.
http://dx.doi.org/10.3109/07853899709113696 [3] Horn-Ross, P.L., Barnes, S., Lee, M., Coward, L., Mandel, J.E., Koo, J., John, E.M. and Smith, M. (2000) Assessing Phytoestrogen Exposure in Epidemiologic Studies: Development of a Database (United States). Cancer Causes & Control, 11, 289-298.
http://dx.doi.org/10.1023/A:1008995606699 [4] Dambroth, M. and Seehuber, R. (1988) Flachs—Züchtung, Anbau, Verarbeitung. Ulmer, Stuttgart. [5] Diepenbrock, W., Fischbeck, G., Heyland, K.U. and Knauer, N. (1999) Spezieller Pflanzenanbau. Ulmer, Stuttgart. [6] Düll, R. and Kutzelnigg, H. (2005) Taschenlexikon der Pflanzen Deutschlands. Quelle & Meyer, Wiebelsheim. [7] Abarzua, S., Szewczyk, M., Gailus, S., Richter, D.-U., Ruth, W., Briese, V. and Piechulla, B. (2007) Effects of Phytoestrogen Extracts from Linum usitatissimum on the Jeg3 Human Trophoblast Tumour Cell Line. Anticancer Research, 27, 2053-2058. [8] Luyengi, L., Suh, N., Fong, H., Pezzuto, J. and Konghorn, D. (1996) A Lignan and Four Terpenoids from Brucea javanica That Induce Differentiation with Cultured HL-60 Promyelotic Leukemia Cells. Phytochemistry, 43, 409-412.
http://dx.doi.org/10.1016/0031-9422(96)00258-0 [9] Matscheski, A., Richter, D.-U., Hartmann, A.M., Effmert, U., Jeschke, U., Kupka, M.S., Abarzua, S., Briese, V., Ruth, W., Kragl, U. and Piechulla, B. (2006) Effects of Phytoestrogen Extracts Isolated from Rye, Green and Yellow Pea Seeds on Hormone Production and Proliferation of Trophoblast Tumor Cells Jeg3. Hormone Research, 65, 276-288.
http://dx.doi.org/10.1159/000092591 [10] Schulten, H.R. and Halket, J.M. (1986) Rapid Characterisation of Biomaterials by Field Ionization. Organic Mass Spectrometry, 21, 613-622.
http://dx.doi.org/10.1002/oms.1210211004 [11] Schulten, H., Simmleit, N. and Mueller, R. (1989) Characterization of Plant Materials by Pyrolysis-Field Ionization Mass Spectrometry: High-Resolution Mass Spectrometry, Time-Resolved High-Resolution Mass Spectrometry, and Curie-Point Pyrolysis-Gas Chromatography/Mass Spectrometry of Spruce Needles. Analytical Chemistry, 61, 221-227.
http://dx.doi.org/10.1021/ac00178a007 [12] Hempfling, R., Simmleit, N. and Schulten, H.R. (1991) Characterization and Chemodynamics of Plant Constituents during Maturation, Senescence and Humus Genesis in Spruce Ecosystems. Biogeochemistry, 13, 27-60.
http://dx.doi.org/10.1007/BF00002875 [13] Hempfling, R. and Schulten, H.R. (1991) Pyrolysis-(Gas Chromatography/) Mass Spectrometry of Agricultural Soils and Their Humic Fractions. Zeitschrift für Pflanzenernährung und Bodenkunde, 154, 425-430.
http://dx.doi.org/10.1002/jpln.19911540606 [14] Booth, N.L., Overk, C.R., Yao, P., Totura, S., Deng, Y., Hedayat, A.S., Bolton, J.L., Pauli, G.F. and Farnsworth, N.R. (2006) Seasonal Variation of Red Clover (Trifolium pratense, L. fabaceae) Isoflavones and Estrogenic Activity. Journal of Agricultural and Food Chemistry, 54, 1277-1282.
http://dx.doi.org/10.1021/jf052927u [15] Godin, B., Lamaudière, S., Agneessens, R., Schmit, T., Goffart, J.P., Stilmant, D., Gerin, P.A. and Delcarte, J. (2013) Chemical Characteristics and Biofuels Potentials of Various Plant Biomasses: Influence of the Harvesting Date. Journal of the Science of Food and Agriculture, 93, 3216-3224.
http://dx.doi.org/10.1002/jsfa.6159 [16] Abarzua, S., Serikawa, T., Szewczyk, M., Richter, D.U., Piechulla, B. and Briese, V. (2012) Antiproliferative Activity of Lignans against the Breast Carcinoma Cell Lines MCF 7 and BT 20. Archives of Gynecology and Obstetrics, 285, 1145-1151.
http://dx.doi.org/10.1007/s00404-011-2120-6 [17] Theil, C., Briese, V., Gerber, B. and Richter, D.U. (2011) The Effects of Different Lignans and Isoflavones, Tested as Aglycones and Glycosides, on Hormone Receptor-Positive and -Negative Breast Carcinoma Cells in Vitro. Archives of Gynecology and Obstetrics, 284, 459-465.
http://dx.doi.org/10.1007/s00404-010-1661-4 [18] Liu, X., Suzuki, N., Santosh Laxmi, Y.R., Okamoto, Y. and Shibutani, S. (2012) Anti-Breast Cancer Potential of Daidzein in Rodents. Life Sciences, 91, 415-419.
http://dx.doi.org/10.1016/j.lfs.2012.08.022 [19] Ono, M., Koga, T., Ueo, H. and Nakano, S. (2012) Effects of Dietary Genistein on Hormone-Dependent Rat Mammary Carcinogenesis Induced by Ethyl Methanesulphonate. Nutrition and Cancer, 64, 1204-1210.
http://dx.doi.org/10.1080/01635581.2012.718035 [20] Lin, C.C., Tsai, Y.L., Ho, C.T. and Teng, S.C. (2008) Determination of the Differential Estrogenicity of Isoflavonoids by E2-ER-ERE Dependent Gene Expression in Recombinant Yeast and MCF-7 Human Breast Cancer Cells. Food Chemistry, 108, 719-726.
http://dx.doi.org/10.1016/j.foodchem.2007.11.020 [21] Mousavi, Y. and Adlercreutz, H. (1992) Enterolactones and Estradiol Inhibit Each Other’s Proliferative Effect on MCF7 Breast Cancer Cells in Culture. The Journal of Steroid Biochemistry and Molecular Biology, 41, 615-619.
http://dx.doi.org/10.1016/0960-0760(92)90393-W [22] Wang, C. and Kurzer, M.S. (1997) Phytoestrogen Concentration Determines Effects on DNA Synthesis in Human Breast Cancer Cells. Nutrition and Cancer, 28, 236-247.
http://dx.doi.org/10.1080/01635589709514582 [23] Welshons, W.V., Murphy, C.S., Koch, R., Calaf, G. and Jordan, V.C. (1987) Stimulation of Breast Cancer Cells in Vitro by the Environmental Estrogen Enterolactone and the Phytoestrogen Equol. Breast Cancer Research and Treatment, 10, 169-175.
http://dx.doi.org/10.1007/BF01810580