Biopolym. Cell. 2008; 24(1):35-40.
Cell Biology
Subcellular localization of S6K1 and S6K2 forms of ribosomal protein S6 kinase in primary monolayer culture of rat thyrocytes
1Khoruzhenko A. I., 1Cherednyk O. V., 1Filonenko V. V.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

The main aim of this work was to determine subcellular localization of S6K1 and S6K2 forms of ribosomal protein S6 kinase in the primary monolayer culture of thyrocytes obtained from undamaged follicles. In the thyroid follicles S6K1, S6K2 have been detected predominantly in the cytoplasm of the cells, however, in the monolayer culture of thyrocytes in the course of follicles outspreading S6K1 and S6K2 have been observed in nuclei as well. Such redistribution of S6K was not directly related to the appearance of proliferating Ki-67 positive cells. At the same time there was a correlation between the appearance of S6K1, S6K2 positive nuclei in monolayer thyrocyte culture and decrease in thyroglobulin content in cultured cells. Thus, the obtained results indicated that the down regulation of thyrocyte functional activity caused by the loss of follicle organization was accompanied by subcellular redistribution of S6K1 and S6K2.
Keywords: ribosomal protein S6 kinase, thyrocytes, primary culture

References

[1] Manning BD. Balancing Akt with S6K: implications for both metabolic diseases and tumorigenesis. J Cell Biol. 2004;167(3):399-403.
[2] Ringel MD, Hayre N, Saito J, Saunier B, Schuppert F, Burch H, Bernet V, Burman KD, Kohn LD, Saji M. Overexpression and overactivation of Akt in thyroid carcinoma. Cancer Res. 2001;61(16):6105-11.
[3] Suh JM, Song JH, Kim DW, Kim H, Chung HK, Hwang JH, Kim JM, Hwang ES, Chung J, Han JH, Cho BY, Ro HK, Shong M. Regulation of the phosphatidylinositol 3-kinase, Akt/protein kinase B, FRAP/mammalian target of rapamycin, and ribosomal S6 kinase 1 signaling pathways by thyroid-stimulating hormone (TSH) and stimulating type TSH receptor antibodies in the thyroid gland. J Biol Chem. 2003;278(24):21960-71.
[4] Coulonval K, Vandeput F, Stein RC, Kozma SC, Lamy F, Dumont JE. Phosphatidylinositol 3-kinase, protein kinase B and ribosomal S6 kinases in the stimulation of thyroid epithelial cell proliferation by cAMP and growth factors in the presence of insulin. Biochem J. 2000;348 Pt 2:351-8.
[5] Medina DL, Santisteban P. Thyrotropin-dependent proliferation of in vitro rat thyroid cell systems. Eur J Endocrinol. 2000;143(2):161-78.
[6] Valovka T, Verdier F, Cramer R, Zhyvoloup A, Fenton T, Rebholz H, Wang ML, Gzhegotsky M, Lutsyk A, Matsuka G, Filonenko V, Wang L, Proud CG, Parker PJ, Gout IT. Protein kinase C phosphorylates ribosomal protein S6 kinase betaII and regulates its subcellular localization. Mol Cell Biol. 2003;23(3):852-63.
[7] Groussin L, Massias JF, Bertagna X, Bertherat J. Loss of expression of the ubiquitous transcription factor cAMP response element-binding protein (CREB) and compensatory overexpression of the activator CREMtau in the human adrenocortical cancer cell line H295R. J Clin Endocrinol Metab. 2000;85(1):345-54.
[8] de Groot RP, Ballou LM, Sassone-Corsi P. Positive regulation of the cAMP-responsive activator CREM by the p70 S6 kinase: an alternative route to mitogen-induced gene expression. Cell. 1994;79(1):81-91.
[9] Richardson CJ, Broenstrup M, Fingar DC, Julich K, Ballif BA, Gygi S, Blenis J. SKAR is a specific target of S6 kinase 1 in cell growth control. Curr Biol. 2004;14(17):1540-9.
[10] Ruvinsky I, Meyuhas O. Ribosomal protein S6 phosphorylation: from protein synthesis to cell size. Trends Biochem Sci. 2006;31(6):342-8.
[11] Kimura T, Van Keymeulen A, Golstein J, Fusco A, Dumont JE, Roger PP. Regulation of thyroid cell proliferation by TSH and other factors: a critical evaluation of in vitro models. Endocr Rev. 2001;22(5):631-56.
[12] Vinals F, Chambard JC, Pouyssegur J. p70 S6 kinase-mediated protein synthesis is a critical step for vascular endothelial cell proliferation. J Biol Chem. 1999;274(38):26776-82.
[13] Kanda S, Miyata Y, Mochizuki Y, Matsuyama T, Kanetake H. Angiopoietin 1 is mitogenic for cultured endothelial cells. Cancer Res. 2005;65(15):6820-7.
[14] Savinska LO, Lyzogubov VV, Usenko VS, Ovcharenko GV, Gorbenko ON, Rodnin MV, Vudmaska MI, Pogribniy PV, Kyyamova RG, Panasyuk GG, Nemazanyy IO, Malets MS, Palchevskyy SS, Gout IT, Filonenko VV. Immunohistochemical analysis of S6K1 and S6K2 expression in human breast tumors. Eksp Onkol. 2004;26(1):24-30.
[15] Westermark K, Nilsson M, Karlsson FA. Effects of interleukin 1 alpha on porcine thyroid follicles in suspension culture. Acta Endocrinol (Copenh). 1990;122(4):505-12.
[16] Goto K, Sasano N, Matoba N. Follicular reconstruction and hormone production by human adenomatous goiter cells in culture. Tohoku J Exp Med. 1982;136(3):285-90.
[17] Bomash N.Y. Morphological diagnosis of thyroid diseases. Moscow: Medicina 1981
[18] Westermark B, Heldin NE, Westermark K. Structural and functional properties of thyroid follicle cells in culture. Acta Physiol Scand Suppl. 1990;592:15-24.
[19] Gartner R. Thyroid growth in vitro. Exp Clin Endocrinol. 1992;100(1-2):32-5.
[20] Massart C, Gibassier J, Genetet N, Raoul ML, Baron M, Le Gall F, Lucas C. Effect of lymphocytes on hormonal secretion by autologous thyrocytes cultured in monolayers. J Mol Endocrinol. 1996;17(3):185-95.
[21] Williams DW, Wynford-Thomas D, Williams ED. Control of human thyroid follicular cell proliferation in suspension and monolayer culture. Mol Cell Endocrinol. 1987;51(1-2):33-40.
[22] Toda S, Yonemitsu N, Hikichi Y, Sugihara H, Koike N. Differentiation of human thyroid follicle cells from normal subjects and Basedow's disease in three-dimensional collagen gel culture. Pathol Res Pract. 1992;188(7):874-82.
[23] Khoruzhenko A.I. New approaches for thyrocyte cultivation in vitro with retention of their follicular organization. Experimental Oncology, 2002; 24 (2):99-104.
[24] Mauchamp J, Mirrione A, Alquier C, Andr? F. Follicle-like structure and polarized monolayer: role of the extracellular matrix on thyroid cell organization in primary culture. Biol Cell. 1998;90(5):369-80.
[25] Dickson JG, Hovsepian S, Fayet G, Lissitzky S. Follicle formation and iodide metabolism in cultures of human thyroid cells. J Endocrinol. 1981;90(1):113-24.
[26] Gerard AC, Denef JF, Colin IM, van den Hove MF. Evidence for processing of compact insoluble thyroglobulin globules in relation with follicular cell functional activity in the human and the mouse thyroid. Eur J Endocrinol. 2004;150(1):73-80.
[27] Antoniak HL, Babych NO, Solohub LI, Snityns'kyi VV. Role of iodothyronine-deiodinase in thyroid hormone mechanisms in animal and human cells. Ukr Biokhim Zh. 2002;74(1):5-18.
[28] Becks GP, Buckingham DK, Wang JF, Phillips ID, Hill DJ. Regulation of thyroid hormone synthesis in cultured ovine thyroid follicles. Endocrinology. 1992;130(5):2789-94.
[29] Lyzogubov V.V., Usenko V.S., Khojaenko Yu.S., Lytvyn D.I., Soldatkina M.A., Rodnin N.V., Filonenko V.V., Pogribniy P.V. Immunohistochemical analysis of p70S6 kinase α in human thyroid tissue upon pathology. Experimental Oncology, 2003; 25 (4), pp. 304-306.
[30] Tepperman J, Tepperman HM. Metabolic and endocrine physiology: an introductory text. Year Book Medical Publishers, 1987 369 p.
[31] Bauer MF, Herzog V. Mini organ culture of thyroid tissue: a new technique for maintaining the structural and functional integrity of thyroid tissue in vitro. Lab Invest. 1988;59(2):281-91.
[32] Chambard M, Verrier B, Gabrion J, Mauchamp J. Polarization of thyroid cells in culture: evidence for the basolateral localization of the iodide "pump" and of the thyroid-stimulating hormone receptor-adenyl cyclase complex. J Cell Biol. 1983;96(4):1172-7.
[33] Panasyuk G, Nemazanyy I, Zhyvoloup A, Bretner M, Litchfield DW, Filonenko V, Gout IT. Nuclear export of S6K1 II is regulated by protein kinase CK2 phosphorylation at Ser-17. J Biol Chem. 2006;281(42):31188-201.
[34] Zhou HY, Wong AS. Activation of p70S6K induces expression of matrix metalloproteinase 9 associated with hepatocyte growth factor-mediated invasion in human ovarian cancer cells. Endocrinology. 2006;147(5):2557-66.
[35] Berven LA, Willard FS, Crouch MF. Role of the p70(S6K) pathway in regulating the actin cytoskeleton and cell migration. Exp Cell Res. 2004;296(2):183-95.
[36] Kanayasu-Toyoda T, Yamaguchi T, Oshizawa T, Kogi M, Uchida E, Hayakawa T. Role of the p70 S6 kinase cascade in neutrophilic differentiation and proliferation of HL-60 cells-a study of transferrin receptor-positive and -negative cells obtained from dimethyl sulfoxide- or retinoic acid-treated HL-60 cells. Arch Biochem Biophys. 2002;405(1):21-31.