Abstract
Genistein, a soy isoflavone, is reported to exert significant beneficial action in several human disorders, which has generated immense interest in the mechanisms underlying its effects on diverse cellular processes. It has anti-proliferative action on many cell types, an effect generally attributed to tyrosine kinase inhibition. In this study, genistein was found to cause total inhibition of [3H]-thymidine incorporation into DNA and a modest reduction in [3H]-proline incorporation into protein in primary cultures of cardiac fibroblasts. The decrease in [3H]-thymidine incorporation was not associated with a decrease in cell proliferation but correlated exactly with low intracellular levels of [3H]-thymidine. Genistein dramatically reduced [3H]-thymidine but not [3H]-proline uptake by these cells in which the equilibrative nucleoside transporter may be the major route of nucleoside uptake. The effect was irreversible and was demonstrable in pulmonary fibroblasts as well. The findings suggest that nucleoside uptake mechanisms may be a novel target of genistein action in cardiac fibroblasts and point to serious limitations in using genistein to assess the role of tyrosine kinase in cell proliferation by the standard technique of [3H]-thymidine incorporation.
References
Akiyama T, Ishida J, Nakagawa S, Ogawara H, Watanabe S, Itoh N, Shibuya M, Fukami Y (1987) Genistein, a specific inhibitor of tyrosine-specific protein kinases. J Biol Chem 262:5592–5596
Yoon HK, Chen K, Baylink DJ, Lau KHW (1998) Differential effects of two protein tyrosine kinase inhibitors, tyrphostin and genistein, on human bone cell proliferation as compared with differentiation. Calcif Tissue Int 63:243–249
Yu JY, Lee JJ, Lim Y, Kim TJ, Jin YR, Sheen YY, Yun YP (2008) Genistein inhibits rat aortic smooth muscle cell proliferation through the induction of p27 kip. J Pharmacol Sci 107:90–98
Eghbali M (1992) Cardiac fibroblasts: function, regulation of gene expression, and phenotypic modulation. Basic Res Cardiol 87(Suppl 2):183–189
Tsuruda T, Jougasaki M, Boerrigter G, Huntley BK, Chen HH, Antonino BD, Lee SC, Larsen AM, Cataliotti A, Burnett JC Jr (2002) Cardiotrophin-1 stimulation of cardiac fibroblast growth roles for glycoprotein 130/leukemia inhibitory factor receptor and the Endothelin Type A receptor. Circ Res 90:128–134
Kumaran C, Shivakumar K (2002) Calcium and superoxide anion-mediated mitogenic action of substance P on cardiac fibroblasts. Am J Physiol Heart Circ Physiol 282:H1855–H1862
Huang M, Wang Y, Cogut SB, Mitchell BS, Graves LM (2003) Inhibition of nucleoside transport by protein kinase inhibitors. J Pharmacol Exp Ther 304:753–760
Cass CE, Young JD, Baldwin SA, Cabrita MA, Graham KA, Griffiths M, Jennings LL, Mackey JR, Ng AM, Ritzel MW, Vickers MF, Yao SY (1999) Nucleoside transporters of mammalian cells. Pharm Biotechnol 12:313–352
Podgorska M, Kocbuch K, Grden M, Szutowicz A, Pawelczyk T (2006) Reduced ability to release adenosine by diabetic rat cardiac fibroblasts due to altered expression of nucleoside transporters. J Physiol 576(1):179–189
He H, Chen L, Zhai M, Chen JZS (2008) Genistein down-regulates the constitutive activation of NF-κb of bone stromal cells in multiple myeloma, leading to suppression of gene expression and proliferation. Drug Dev Res 69:219–225
Thasni KA, Rojini G, Nair RS, Ratheeshkumar T, Babu MS, Srinivas G, Banerji A, Priya S (2008) Genistein induces apoptosis in ovarian cancer cells via different molecular pathways depending on breast cancer susceptibility gene-1 (BRCA1) status. Eur J Pharmacol 588:158–164. doi:10.1016/j.ejphar.2008.04.041
Weber G (1983) Biochemical strategy of cancer cells and the design of chemotherapy: G.H.A.Clowes Memorial Lecture. Cancer Res 43:3466–3492
Vera JC, Reyes AM, Carcamo JG, Velasquez FV, Rivas CI, Zhang RH, Strobel P, Iribarren R, Scher HI, Slebe JC, David WG (1996) Genistein is a natural inhibitor of hexose and dehydroascorbic acid transport through the glucose transporter, GLUT1. J Biol Chem 27:8719–8724
Baldwin SA, Mackey JR, Cass CE, Young JD (1995) Nucleoside transporters: molecular biology and implications for therapeutic development. Mol Med Today 5:216–224
Plagemann PG, Wohlhueter RM, Woffendin C (1988) Nucleoside and nucleobase transport in animal cells. Biochem Biophys Acta 947:405–443
Mubagwa K, Flameng W (2001) Adenosine, adenosine receptors and myocardial protection: an updated overview. Cardivasc Res 52:25–39
Acknowledgments
This work was supported by a research grant to SK from the Department of Biotechnology, Government of India. MSP gratefully acknowledges the Junior Research Fellowship from the Life Sciences Research Board, DRDO, Government of India. The authors thank the Director, SCTIMST, for facilities.
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Pillai, M.S., Shivakumar, K. Genistein abolishes nucleoside uptake by cardiac fibroblasts. Mol Cell Biochem 332, 121–125 (2009). https://doi.org/10.1007/s11010-009-0181-7
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DOI: https://doi.org/10.1007/s11010-009-0181-7