The polyphenol curcumin (diferuloylmethane) is the active componenet of the spice plant Curcuma longa and has been shown to exert multiple actions on mammalian cells. We have studied its effect on folliculostellate (FS) TtT/GF mouse pituitary cells, representative of a multifunctional, endocrine inactive cell type of the anterior pituitary. Proliferation of TtT/GF cells was inhibited by curcumin in a monolayer cell culture and in the colony formation assay in soft agar. Fluorescence-activated cell-sorting (FACS) analysis demonstrated curcumin-induced cell cycle arrest at G2/M accompanied by inhibition of cyclin D1 protein expression. Curcumin had a small effect on necrosis of TtT/GF cells, but it mainly stimulated apoptosis as demonstrated by FACS analysis (Annexin V-fluorescein isothiocyannate/7-aminoactinomycin D staining). Curcumin-induced apoptosis involved suppression of Bcl-2, stimulation of cleaved caspase-3 and induction of DNA fragmentation. Functional studies on FS cell-derived compounds showed that curcumin inhibited mRNA synthesis and release of angiogenic vascular endothelial growth factor-A (VEGF-A). Immune-like functions of FS cells were impaired since curcumin downregulated Toll-like receptor 4, reduced nuclear factor-ĸB expression and suppressed bacterial endotoxin-induced interleukin-6 (IL-6) secretion. The inhibitory action of curcumin on VEGF-A and IL-6 production was also found in primary rat pituitary cell cultures, in which FS cells are the only source of these proteins. The observed effects of curcumin on FS cell growth, apoptosis and functions may have therapeutic consequences for the intrapituitary regulation of hormone production and release as well as for pituitary tumor pathogenesis.

Keywords:
Curcumin, Folliculostellate pituitary cells, Proliferation, Cell cycle, Apoptosis, Vascular endothelial growth factor, Interleukin-6, Tlr4, Nuclear factor-ĸB
1.
Rinehart JF, Farquhar MG: Electron microscopic studies of the anterior pituitary gland. J Histochem Cytochem 1953;1:93–113.
2.
Inoue K, Couch EF, Takano K, Ogawa S: The structure and function of folliculo-stellate cells in the anterior pituitary gland. Arch Histol Cytol 1999;62:205–218.
3.
Allaerts W, Vankelecom H: History and perspectives of pituitary folliculo-stellate cell research. Eur J Endocrinol 2005;153:1–12.
4.
Fauquier T, Guerineau NC, McKinney RA, Bauer K, Mollard P: Folliculostellate cell network: a route for long-distance communication in the anterior pituitary. Proc Natl Acad Sci USA 2001;98:8891–8896.
5.
Fauquier T, Lacampagne A, Travo P, Bauer K, Mollard P: Hidden face of the anterior pituitary. Trends Endocrinol Metab 2002:13;304–309.
6.
Morand I, Fonlupt P, Guerrier A, Trouillas J, Calle A, Remy C, Rousset B, Munari-Silem Y: Cell-to-cell communication in the anterior pituitary: evidence for gap junction-mediated exchanges between endocrine cells and folliculostellate cells. Endocrinology 1996;137:3356–3367.
7.
Lewis BM, Pexa A, Francis K, Verma V, McNicol AM, Scanlon M, Deussen A, Evans WH, Rees A, Ham J: Adenosine stimulates connexin 43 expression and gap junctional communication in pituitary folliculostellate cells. FASEB J 2006;20:2585–2587.
8.
Meilleur MA, Akpovi CD, Pelletier RM, Vitale ML: Tumor necrosis factor-α-induced anterior pituitary folliculostellate TtT/GF cell uncoupling is mediated by connexin 43 dephosphorylation. Endocrinology 2007;148:5913–5924.
9.
Aoki A, de Gaisan EO, Pasolli HA, Torres AI: Disposal of cell debris from surplus lactotrophs of pituitary gland. Exp Clin Endocrinol Diabetes 1996;104:256–262.
10.
Luziga C, Yamamoto Y, Horii Y, Mbassa G, Mamba K: Phagocytotic removal of apoptotic endocrine cells by folliculostellate cells and its functional implications in clusterin accumulation in pituitary colloids in helmeted guinea fowl (Numida meleagris). Acta Histochem 2006;108:69–80.
11.
Allaerts W, Jeucken PHM, Debets R, Hoefakker S, Claassen E, Drexhage HA: Heterogeneity of pituitary folliculo-stellate cells: implications for IL-6 production and accessory function in vitro. J Neuroendocrinol 1997;9:43–53.
12.
Fauquier T, Rizzoti K, Dattani M, Lovell-Bage R, Robinson IC: SOX2-expressing progenitor cells generate all of the major cell types in the adult mouse pituitary gland. Proc Natl Acad Sci USA 2008;105:2907–2912.
13.
Gospodarowicz D, Abraham JA, Schilling J: Isolation and characterization of a vascular cell mitogen produced by pituitary-derived folliculostellate cells. Proc Natl Acad Sci USA 1989;6:7311–7315.
14.
Vankelecom H, Carmeliet P, Van Damme J, Billiau A, Denef C: Production of interleukin-6 by folliculo-stellate cells of the anterior pituitary gland in a histiotypic cell aggregate culture system. Neuroendocrinology 1989;49:102–106.
15.
Renner U, De Santana EC, Gerez J, Fröhlich B, Haedo M, Pereda MP, Onofri C, Stalla GK, Arzt E: Intrapituitary expression and regulation of the gp130 cytokine interleukin-6 and its implication in pituitary physiology and pathophysiology. Ann N Y Acad Sci 2009;1153:89–97.
16.
Ferrara N: Vascular endothelial growth factor: basic science and clinical progress. Endocr Rev 2004;25:581–611.
17.
Lohrer P, Gloddek J, Nagashima AC, Korali Z, Hopfner U, Pereda MP, Arzt E, Stalla GK, Renner U: Lipopolysaccharide directly stimulates the intrapituitary interleukin-6 production by folliculostellate cells via specific receptors and the p38α mitogen-activated protein kinase/nuclear factor-ĸB pathway. Endocrinology 2000;141:4457–4465.
18.
Gloddek J, Lohrer P, Stalla J, Arzt E, Stalla GK, Renner U: The intrapituitary stimulatory effect of lipopolysaccharide on ACTH secretion is mediated by paracrine-acting IL-6. Exp Clin Endocrinol Diabetes 2001;109:410–415.
19.
Aggarwal BB, Sundaram C, Malani N, Ichikawa H: Curcumin: the Indian solid gold. Adv Exp Med Biol 2007;595:1–75.
20.
Hatcher H, Planalp R, Cho J, Torti FM, Torti SV: Curcumin: from ancient medicine to current clinical trials. Cell Mol Life Sci 2008;65:1631–1652.
21.
Shishodia S, Chaturvedi MM, Aggarwal BB: Role of curcumin in cancer therapy. Current Probl Cancer 2007;31:243–305.
22.
Singh S: From exotic spice to modern drug? Cell 2007;130:765–768.
23.
Sa G, Das T: Anti cancer effects of curcumin: cycle of life and death. Cell Div 2008;3:1–14.
24.
Shankar S, Ganapathy S, Chen Q, Srivastava RK: Curcumin sensitizes TRAIL-resistant xenografts: molecular mechanisms of apoptosis, metastasis and angiogenesis. Mol Cancer 2008;7:16.
25.
Corson TW, Crews CM: Molecular understanding and modern application of traditional medicines: triumphs and trials. Cell 2007;130:769–774.
26.
Dhillon N, Aggarwal BB, Newman RA, Wolff RA, Kunnumakkara AB, Abbruzzese JL, Ng CS, Badmaev V, Kurzrock R: Phase II trial of curcumin in patients with advanced pancreatic cancer. Clin Cancer Res 2008;14:4491–4499.
27.
Johnson JJ, Mukhtar H: Curcumin for chemoprevention of colon cancer. Cancer Lett 2007;255:170–181.
28.
Kunnumakkara AB, Anand P, Aggarwal BB: Curcumin inhibits proliferation, invasion, angiogenesis and metastasis of different cancers through interaction with multiple cell signalling proteins. Cancer Lett 2008;269:199–225.
29.
Carroll CE, Ellersieck MR, Hyder SM: Curcumin inhibits MPA-induced secretion of VEGF from T47-D human breast cancer cells. Menopause 2008;15:570–574.
30.
Chadalapaka G, Jutooru I, Chintharlapalli S, Papineni S, Smith R 3rd, Li X, Safe S: Curcumin decreases specificity protein expression in bladder cancer cells. Cancer Res 2008;68:5345–5354.
31.
Sandur SK, Ichikawa H, Pandey MK, Kunnumakkara AB, Sung B, Sethi G, Aggarwal BB: Role of pro-oxidants and antioxidants in the anti-inflammatory and apoptotic effects of curcumin (diferuloylmethane). Free Radic Biol Med 2007;43:568–580.
32.
Miller M, Chen S, Woodliff J, Kansra S: Curcumin (diferuloylmethane) inhibits cell proliferation, induces apoptosis, and decreases hormone levels and secretion in pituitary tumor cells. Endocrinology 2008;149:4158–4167.
33.
Inoue K, Matsumoto H, Koyama C, Shibata K, Nakazato Y, Ito A: Establishment of a folliculostellate-like cell line from a murine thyrotropic pituitary tumor. Endocrinology 1992;131:3110–3116.
34.
Renner U, Newton CJ, Pagotto U, Sauer J, Arzt E, Stalla GK: Involvement of interleukin-1 and interleukin-1 receptor antagonist in rat pituitary cell growth regulation. Endocrinology 1995;136:3186–3193.
35.
Onofri C, Theodoropoulou M, Losa M, Uhl E, Lange M, Arzt E, Stalla GK, Renner U: Localization of vascular endothelial growth factor (VEGF) receptors in normal and adenomatous pituitaries: detection of a non-endothelial function of VEGF in pituitary tumors. J Endocrinol 2006;191:249–261.
36.
Nolan LA, Kavanagh E, Lightman SL, Levy A: Anterior pituitary cell population control: basal turnover and the effects of adrenalectomy and dexamethasone treatment. J Neuroendocrinol 1998;10:207–215.
37.
Levy A: Physiological implications of pituitary trophic activity. J Endocrinol 2002;174:147–155.
38.
Besedovsky HO, del Rey A: Immune-neuro-endocrine interactions: facts and hypotheses. Endocr Rev 1996;17:64–102.
39.
Herkenham M: Folliculo-stellate (FS) cells of the anterior pituitary mediate interactions between the endocrine and immune systems. Endocrinology 2005;146:33–34.
40.
Gautam SC, Gao X, Dulchavsky S: Immunomodulation by curcumin. Adv Exp Med Biol 2007;595:321–341.
41.
Lubbad A, Oriowo MA, Kahn I: Curcumin attenuates inflammation through inhibition of TLR-4 receptor in experimental colitis. Mol Cell Biochem 2009;322:127–135.
42.
Harper SJ, Bates DO: VEGF-A splicing: the key to anti-angiogenic therapeutics? Nat Rev Cancer 2008;8:880–887.
43.
Jabbour HN, Boddy SC, Lincoln GA: Pattern of localisation of expression of vascular endothelial growth factor and its receptor flt-1 in the ovine pituitary gland: expression is independent of hypothalamic control. Mol Cell Endocrinol 1997;134:91–100.
44.
Arzt E, Buric R, Stelzer G, Stalla J, Sauer J, Renner U, Stalla GK: Interleukin involvement in anterior pituitary cell growth regulation: effects of IL-2 and IL-6. Endocrinology 1993;132:459–467.
45.
Hori S, Hayashi N, Fukuoda J, Kurimoto M, Hamada H, Miyajima K, Nagai S, Endo S: Folliculostellate cell tumor in the pituitary gland. Neuropathology 2009;29:78–80.
46.
Roncaroli F, Scheithauer BW, Cenacchi G, Horvath E, Kovacs K, Lloyd RV, Abell-Aleff P, Santi M, Yates AJ: ‘Spindle cell oncocytoma’ of the adenohypophysis. A tumor of folliculostellate cells? Am J Surg Pathol 2002;26:1048–1055.
47.
Iwaki T, Kondo A, Takeshita I, Nakagaki H, Kitamura K, Tateishi J: Proliferating potential of folliculo-stellate cells in human pituitary adenomas. Immunohistochemical and electron microscopic analysis. Acta Neuropathol 1986;71:233–242.
48.
Höfler H, Walter GF, Denk H: Immunohistochemistry of folliculo-stellate cells in normal human adenohypopheses and in pituitary adenomas. Acta Neuropathol 1984;65:35–40.
49.
Farnoud MR, Kujas M, Derome P, Racadot J, Peillon F, Li JY: Interactions between normal and tumoral tissues at the boundary of human pituitary adenomas. Virchows Arch 1994;424:75–82.
50.
Yamashita M, Qian ZR, Sano T, Horvath E, Kovacs K: Immunohistochemical study on so-called follicular cells and folliculostellate cells in the human adenohypophysis. Pathol Int 2005;55:244–247.
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