Journal of Advances in Molecular Biology

Estrogen Activation of CaM Kinases and Transcription Is Blocked by Vitamin D in MCF-7 Breast Cancer Cells

Download PDF (1010.8 KB) PP. 129 - 147 Pub. Date: December 14, 2017

DOI: 10.22606/jamb.2017.13001

Author(s)

John M. Schmitt^*
From the Biology Department, George Fox University, 414 N. Meridian St, Newberg, OR 97132
Jessica Magill
From the Biology Department, George Fox University, 414 N. Meridian St, Newberg, OR 97132
Amanda Ankeny
From the Biology Department, George Fox University, 414 N. Meridian St, Newberg, OR 97132
Renee Geck
From the Biology Department, George Fox University, 414 N. Meridian St, Newberg, OR 97132
Jessica Milligan
From the Biology Department, George Fox University, 414 N. Meridian St, Newberg, OR 97132
Hannah McFarland
From the Biology Department, George Fox University, 414 N. Meridian St, Newberg, OR 97132
Erica Rice
From the Biology Department, George Fox University, 414 N. Meridian St, Newberg, OR 97132

Abstract

Calcium/calmodulin-dependent protein kinase (CaM Kinase) proteins are targets of hormones and growth factors and regulate cancer cell growth, apoptosis and migration. The hormone estrogen (E2) utilizes CaM Kinases to activate the Extracellular-signal regulated kinase (ERK) leading to MCF-7 breast cancer cell growth. The hormone Vitamin D (Vit D) may inhibit breast cancer cell growth however the cellular mechanisms of Vit D action remain to be elucidated. Within the present study we provide data that E2 stimulation of MCF-7 cells activates CaM Kinase Kinase (CaM KK), CaM KI, and ERK and the transcription factors Elk-1 and SRF. E2 treatment of MCF-7 cells potently stimulated Elk-1/SRF directed transcription of SRE-luciferase reporters. E2 activation of ERK, Elk-1, SRF and SRE-dependent luciferase activities were blocked by treating cells with the CaM KK inhibitor, STO-609, and the ERK inhibitor, U0126. Moreover, siRNAs directed against CaM KK and ERK blocked transcription factor phosphorylation and luciferase activity. Treatment of cells with Vit D, the hormone Epinephrine (Epi), or Forskolin, prevented E2 activation of CaM KK and ERK. Interestingly, Vit D promoted a PKA-dependent phosphorylation and inhibition of CaM KK as well as its association with 14-3-3. Epi and Vit D treatment of cells blocked the ability of E2 to active Elk-1 and SRE-luciferase activity. This data suggests an important role for CaM KK and ERK in regulating transcription downstream of E2 in MCF-7 cells. Our results also suggest that Vit D treatment of MCF-7 cells utilizes a unique PKA-dependent mechanism to block E2 activation of CaM KK, ERK and transcription.

Keywords

Estrogen (E2); CaM KK, calcium/calmodulin-dependent protein kinase kinase; ERK, extracellular-regulated protein kinase; vitamin D (Vit D); PKA, protein kinase A ; Elk-1

References

[1] Fuhrman, B.J., et al., Sunlight, polymorphisms of vitamin D-related genes and risk of breast cancer. Anticancer research, 2013. 33(2): p. 543-51.

[2] Garland, F.C., et al., Geographic variation in breast cancer mortality in the United States: a hypothesis involving exposure to solar radiation. Prev Med, 1990. 19(6): p. 614-22.

[3] Shao, T., P. Klein, and M.L. Grossbard, Vitamin D and breast cancer. The oncologist, 2012. 17(1): p. 36-45.

[4] Colston, K.W., et al., Vitamin D status and breast cancer risk. Anticancer research, 2006. 26(4A): p. 2573-80.

[5] Tangpricha, V., et al., Vitamin D deficiency enhances the growth of MC-26 colon cancer xenografts in Balb/c mice. The Journal of nutrition, 2005. 135(10): p. 2350-4.

[6] Ooi, L.L., et al., Vitamin D deficiency promotes growth of MCF-7 human breast cancer in a rodent model of osteosclerotic bone metastasis. Bone, 2010. 47(4): p. 795-803.

[7] Ooi, L.L., et al., Vitamin D deficiency promotes human breast cancer growth in a murine model of bone metastasis. Cancer research, 2010. 70(5): p. 1835-44.

[8] Alimirah, F., et al., Functional significance of vitamin D receptor FokI polymorphism in human breast cancer cells. PLoS One, 2011. 6(1): p. e16024.

[9] Richard, C.L., et al., Involvement of 1,25D3-MARRS (membrane associated, rapid response steroid-binding), a novel vitamin D receptor, in growth inhibition of breast cancer cells. Exp Cell Res, 2010. 316(5): p. 695-703.

[10] Boyan, B.D., et al., Plasma membrane requirements for 1alpha,25(OH)2D3 dependent PKC signaling in chondrocytes and osteoblasts. Steroids, 2006. 71(4): p. 286-90.

[11] Boland, R.L., VDR activation of intracellular signaling pathways in skeletal muscle. Mol Cell Endocrinol, 2011. 347(1-2): p. 11-6.

[12] Larriba, M.J., et al., Interaction of vitamin D with membrane-based signaling pathways. Front Physiol, 2014. 5: p. 60.

[13] Slominski, A.T., et al., Novel vitamin D hydroxyderivatives inhibit melanoma growth and show differential effects on normal melanocytes. Anticancer Res, 2012. 32(9): p. 3733-42.

[14] Slominski, A.T., et al., 20-Hydroxyvitamin D2 is a noncalcemic analog of vitamin D with potent antiproliferative and prodifferentiation activities in normal and malignant cells. Am J Physiol Cell Physiol, 2011. 300(3): p. C526-41.

[15] Kittaka, A., et al., Potent 19-norvitamin D analogs for prostate and liver cancer therapy. Future Med Chem, 2012. 4(16): p. 2049-65.

[16] Chiang, K.C., et al., MART-10, a New Generation of Vitamin D Analog, Is More Potent than 1alpha,25-Dihydroxyvitamin D(3) in Inhibiting Cell Proliferation and Inducing Apoptosis in ER+ MCF-7 Breast Cancer Cells. Evid Based Complement Alternat Med, 2012. 2012: p. 310872.

[17] Karlsson, S., et al., Vitamin D and prostate cancer: the role of membrane initiated signaling pathways in prostate cancer progression. J Steroid Biochem Mol Biol, 2010. 121(1-2): p. 413-6.

[18] Masuyama, R., et al., Vitamin D receptor in chondrocytes promotes osteoclastogenesis and regulates FGF23 production in osteoblasts. J Clin Invest, 2006. 116(12): p. 3150-9.

[19] Wang, Y., J. Zhu, and H.F. DeLuca, Identification of the vitamin D receptor in osteoblasts and chondrocytes but not osteoclasts in mouse bone. J Bone Miner Res, 2014. 29(3): p. 685-92.

[20] Cordes, T., et al., Modulation of MAPK ERK1 and ERK2 in VDR-positive and -negative breast cancer cell lines. Anticancer Res, 2006. 26(4A): p. 2749-53.

[21] Cordes, T., et al., Expression of splice variants of 1alpha-hydroxylase in mcf-7 breast cancer cells. J Steroid Biochem Mol Biol, 2007. 103(3-5): p. 326-9.

[22] Fischer, D., et al., 25-Hydroxyvitamin D3 1alpha-hydroxylase splice variants in breast cell lines MCF-7 and MCF-10. Cancer Genomics Proteomics, 2007. 4(4): p. 295-300.

[23] Berg, J.P. and E. Haug, Vitamin D: a hormonal regulator of the cAMP signaling pathway. Crit Rev Biochem Mol Biol, 1999. 34(5): p. 315-23.

[24] Boyan, B.D. and Z. Schwartz, Rapid vitamin D-dependent PKC signaling shares features with estrogen-dependent PKC signaling in cartilage and bone. Steroids, 2004. 69(8-9): p. 591-7.

[25] Carlberg, C. and M.J. Campbell, Vitamin D receptor signaling mechanisms: integrated actions of a well-defined transcription factor. Steroids, 2013. 78(2): p. 127-36.

[26] Gniadecki, R., Nongenomic signaling by vitamin D: a new face of Src. Biochem Pharmacol, 1998. 56(10): p. 1273-7.

[27] Sergeev, I.N., Vitamin D and cellular Ca2+ signaling in breast cancer. Anticancer Res, 2012. 32(1): p. 299-302.

[28] Brosseau, C.M., G. Pirianov, and K.W. Colston, Involvement of stress activated protein kinases (JNK and p38) in 1,25 dihydroxyvitamin D3-induced breast cell death. Steroids, 2010. 75(13-14): p. 1082-8.

[29] Capiati, D.A., et al., Inhibition of serum-stimulated mitogen activated protein kinase by 1alpha,25(OH)2-vitamin D3 in MCF-7 breast cancer cells. J Cell Biochem, 2004. 93(2): p. 384-97.

[30] Visram, H. and P.A. Greer, 17beta-estradiol and tamoxifen stimulate rapid and transient ERK activationin MCF-7 cells via distinct signaling mechanisms. Cancer Biol Ther, 2006. 5(12): p. 1677-82.

[31] Schmitt, J.M., et al., ERK activation and cell growth require CaM kinases in MCF-7 breast cancer cells. Mol Cell Biochem, 2010. 335(1-2): p. 155-71.

[32] Liu, J.F., et al., FGF-2 and TPA induce matrix metalloproteinase-9 secretion in MCF-7 cells through PKC activation of the Ras/ERK pathway. Biochem Biophys Res Commun, 2002. 293(4): p. 1174-82.

[33] Muscella, A., et al., PKC-zeta is required for angiotensin II-induced activation of ERK and synthesis of C-FOS in MCF-7 cells. J Cell Physiol, 2003. 197(1): p. 61-8.

[34] Rieber, M. and M. Strasberg-Rieber, p53 inactivation decreases dependence on estrogen/ERK signalling for proliferation but promotes EMT and susceptility to 3-bromopyruvate in ERalpha+ breast cancer MCF-7 cells. Biochem Pharmacol, 2014. 88(2): p. 169-77.

[35] Aksamitiene, E., et al., Prolactin-stimulated activation of ERK1/2 mitogen-activated protein kinases is controlled by PI3-kinase/Rac/PAK signaling pathway in breast cancer cells. Cell Signal, 2011. 23(11): p. 1794-805.

[36] Improta-Brears, T., et al., Estrogen-induced activation of mitogen-activated protein kinase requires mobilization of intracellular calcium. Proc Natl Acad Sci U S A, 1999. 96(8): p. 4686-91.

[37] Thomas, W., et al., Estrogen induces phospholipase A2 activation through ERK1/2 to mobilize intracellular calcium in MCF-7 cells. Steroids, 2006. 71(3): p. 256-65.

[38] Wayman, G.A., et al., Analysis of CaM-kinase signaling in cells. Cell Calcium, 2011.

[39] Colomer, J. and A.R. Means, Physiological roles of the Ca2+/CaM-dependent protein kinase cascade in health and disease. Subcell Biochem, 2007. 45: p. 169-214.

[40] Skelding, K.A., J.A. Rostas, and N.M. Verrills, Controlling the cell cycle: the role of calcium/calmodulin-stimulated protein kinases I and II. Cell Cycle, 2011. 10(4): p. 631-9.

[41] Racioppi, L. and A.R. Means, Calcium/calmodulin-dependent protein kinase kinase 2: roles in signaling and pathophysiology. J Biol Chem, 2012. 287(38): p. 31658-65.

[42] Wayman, G.A., et al., An activity-regulated microRNA controls dendritic plasticity by down-regulating p250GAP. Proc Natl Acad Sci U S A, 2008. 105(26): p. 9093-8.

[43] Chang, F., et al., Signal transduction mediated by the Ras/Raf/MEK/ERK pathway from cytokine receptors to transcription factors: potential targeting for therapeutic intervention. Leukemia, 2003. 17(7): p. 1263-93.

[44] Wiegert, J.S. and H. Bading, Activity-dependent calcium signaling and ERK-MAP kinases in neurons: a link to structural plasticity of the nucleus and gene transcription regulation. Cell Calcium, 2011. 49(5): p. 296-305.

[45] Zhang, H.M., et al., Mitogen-induced recruitment of ERK and MSK to SRE promoter complexes by ternary complex factor Elk-1. Nucleic Acids Res, 2008. 36(8): p. 2594-607.

[46] Gille, H., et al., ERK phosphorylation potentiates Elk-1-mediated ternary complex formation and transactivation. Embo J, 1995. 14(5): p. 951-62.

[47] Cruzalegui, F.H., E. Cano, and R. Treisman, ERK activation induces phosphorylation of Elk-1 at multiple S/T-P motifs to high stoichiometry. Oncogene, 1999. 18(56): p. 7948-57.

[48] Stork, P.J. and J.M. Schmitt, Crosstalk between cAMP and MAP kinase signaling in the regulation of cell proliferation. Trends Cell Biol, 2002. 12(6): p. 258-66.

[49] Ligumsky, H., et al., The peptide-hormone glucagon-like peptide-1 activates cAMP and inhibits growth of breast cancer cells. Breast Cancer Res Treat, 2012. 132(2): p. 449-61.

[50] Davare, M.A., et al., Inhibition of calcium/calmodulin-dependent protein kinase kinase by protein 14-3-3. J Biol Chem, 2004. 279(50): p. 52191-9.

[51] Wayman, G.A., et al., Activity-dependent dendritic arborization mediated by CaM-kinase I activation and enhanced CREB-dependent transcription of Wnt-2. Neuron, 2006. 50(6): p. 897-909.

[52] Sato, K., et al., Regulation of osteoclast differentiation and function by the CaMK-CREB pathway. Nat Med, 2006. 12(12): p. 1410-6.

[53] Schmitt, J.M., et al., Calcium activation of ERK mediated by calmodulin kinase I. J Biol Chem, 2004. 279(23): p. 24064-72.

[54] Enslen, H., et al., Regulation of mitogen-activated protein kinases by a calcium/calmodulin-dependent protein kinase cascade. Proc Natl Acad Sci U S A, 1996. 93(20): p. 10803-8.

[55] Rodriguez-Mora, O.G., et al., Calcium/calmodulin-dependent kinase I and calcium/calmodulin-dependent kinase kinase participate in the control of cell cycle progression in MCF-7 human breast cancer cells. Cancer Res, 2005. 65(12): p. 5408-16.

[56] Tokumitsu, H., et al., STO-609, a specific inhibitor of the Ca(2+)/calmodulin-dependent protein kinase kinase. J Biol Chem, 2002. 277(18): p. 15813-8.

[57] Tokumitsu, H., et al., A single amino acid difference between alpha and beta Ca2+/calmodulin-dependent protein kinase kinase dictates sensitivity to the specific inhibitor, STO-609. J Biol Chem, 2003. 278(13): p. 10908-13.

[58] Bebien, M., et al., Immediate-early gene induction by the stresses anisomycin and arsenite in human osteosarcoma cells involves MAPK cascade signaling to Elk-1, CREB and SRF. Oncogene, 2003. 22(12): p. 1836-47.

[59] Aplin, A.E., et al., Integrin-mediated adhesion regulates ERK nuclear translocation and phosphorylation of Elk-1. J Cell Biol, 2001. 153(2): p. 273-82.

[60] Schmitt, J.M., et al., CaM kinase control of AKT and LNCaP cell survival. J Cell Biochem, 2012. 113(5): p. 1514-26.

[61] Evans, E.L., et al., Dimer formation and conformational flexibility ensure cytoplasmic stability and nuclear accumulation of Elk-1. Nucleic Acids Res, 2011. 39(15): p. 6390-402.

[62] Wayman, G.A., H. Tokumitsu, and T.R. Soderling, Inhibitory cross-talk by cAMP kinase on the calmodulin-dependent protein kinase cascade. J Biol Chem, 1997. 272(26): p. 16073-6.

[63] Matsushita, M. and A.C. Nairn, Inhibition of the Ca2+/calmodulin-dependent protein kinase I cascade by cAMP-dependent protein kinase. J Biol Chem, 1999. 274(15): p. 10086-93.

[64] Bellido, T., et al., Evidence for the participation of protein kinase C and 3',5'-cyclic AMP-dependent protein kinase in the stimulation of muscle cell proliferation by 1,25-dihydroxy-vitamin D3. Mol Cell Endocrinol, 1993. 90(2): p. 231-8.

[65] Anderson, K.A., et al., Hypothalamic CaMKK2 contributes to the regulation of energy balance. Cell Metab, 2008. 7(5): p. 377-88.

[66] Hell, J.W., CaMKII: claiming center stage in postsynaptic function and organization. Neuron, 2014. 81(2): p. 249-65.

[67] Shonesy, B.C., et al., CaMKII: a molecular substrate for synaptic plasticity and memory. Prog Mol Biol Transl Sci, 2014. 122: p. 61-87.

[68] Monaco, S., et al., A novel crosstalk between calcium/calmodulin kinases II and IV regulates cell proliferation in myeloid leukemia cells. Cell Signal, 2015. 27(2): p. 204-14.

[69] Lin, F., et al., The camKK2/camKIV relay is an essential regulator of hepatic cancer. Hepatology, 2015.

[70] Su, R., et al., MiR-181 family: regulators of myeloid differentiation and acute myeloid leukemia as well as potential therapeutic targets. Oncogene, 2014. 0.

[71] Racioppi, L., CaMKK2: a novel target for shaping the androgen-regulated tumor ecosystem. Trends Mol Med, 2013. 19(2): p. 83-8.

[72] Davare, M.A., T. Saneyoshi, and T.R. Soderling, Calmodulin-kinases regulate basal and estrogen stimulated medulloblastoma migration via Rac1. J Neurooncol, 2011. 104(1): p. 65-82.

[73] Zhang, Y.H., et al., Polymorphism rs7214723 in CAMKK1 and lung cancer risk in Chinese population. Tumour Biol, 2013. 34(5): p. 3147-52.

[74] Daft, P.G., et al., Alpha-CaMKII plays a critical role in determining the aggressive behavior of human osteosarcoma. Mol Cancer Res, 2013. 11(4): p. 349-59.

[75] Iglewski, M. and S.R. Grant, Urotensin II-induced signaling involved in proliferation of vascular smooth muscle cells. Vasc Health Risk Manag, 2010. 6: p. 723-34.

[76] Ma, Z., et al., Growth inhibition of human gastric adenocarcinoma cells in vitro by STO-609 is independent of calcium/calmodulin-dependent protein kinase kinase-beta and adenosine monophosphate-activated protein kinase. Am J Transl Res, 2016. 8(2): p. 1164-71.

[77] Deshmukh, R.R. and Q.P. Dou, Proteasome inhibitors induce AMPK activation via CaMKKbeta in human breast cancer cells. Breast Cancer Res Treat, 2015. 153(1): p. 79-88.

[78] Popovics, P., et al., Targeting the 5'-AMP-activated protein kinase and related metabolic pathways for the treatment of prostate cancer. Expert Opin Ther Targets, 2015. 19(5): p. 617-32.

[79] Mamaeva, O.A., et al., Calcium/calmodulin-dependent kinase II regulates notch-1 signaling in prostate cancer cells. J Cell Biochem, 2009. 106(1): p. 25-32.

[80] Chai, S., et al., Ca2+/calmodulin-dependent protein kinase IIgamma enhances stem-like traits and tumorigenicity of lung cancer cells. Oncotarget, 2015. 6(18): p. 16069-83.

[81] Hoffman, A., et al., Dephosphorylation of CaMKII at T253 controls the metaphase-anaphase transition. Cell Signal, 2014. 26(4): p. 748-56.

[82] Chen, S., et al., Fucoidan induces cancer cell apoptosis by modulating the endoplasmic reticulum stress cascades. PLoS One, 2014. 9(9): p. e108157.

[83] Rodriguez-Mora, O.G., et al., Inhibition of the CaM-kinases augments cell death in response to oxygen radicals and oxygen radical inducing cancer therapies in MCF-7 human breast cancer cells. Cancer Biol Ther, 2006. 5(8): p. 1022-30.

[84] Hidaka, H., M. Hagiwara, and H. Tokumitsu, Novel and selective inhibitors of CaM-kinase II and other calmodulin-dependent enzymes. Adv Exp Med Biol, 1990. 269: p. 159-62.

[85] Zheng, G., et al., 14-3-3sigma regulation by p53 mediates a chemotherapy response to 5-fluorouracil in MCF-7 breast cancer cells via Akt inactivation. FEBS Lett, 2012. 586(2): p. 163-8.

[86] Urano, T., et al., Efp targets 14-3-3 sigma for proteolysis and promotes breast tumour growth. Nature, 2002. 417(6891): p. 871-5.

[87] Onouchi, T., et al., Regulation of Ca(2+)/calmodulin-dependent protein kinase phosphatase (CaMKP/PPM1F) by protocadherin-gammaC5 (Pcdh-gammaC5). Arch Biochem Biophys, 2015. 585: p. 109-20.

[88] Saneyoshi, T., et al., Activity-dependent synaptogenesis: regulation by a CaM-kinase kinase/CaM-kinase I/betaPIX signaling complex. Neuron, 2008. 57(1): p. 94-107.

[89] O'Leary, H., et al., Nuclear targeting of the CaMKII anchoring protein alphaKAP is regulated by alternative splicing and protein kinases. Brain Res, 2006. 1086(1): p. 17-26.

[90] Woolfrey, K.M. and M.L. Dell'Acqua, Coordination of Protein Phosphorylation and Dephosphorylation in Synaptic Plasticity. J Biol Chem, 2015. 290(48): p. 28604-12.

[91] Hashimoto, Y., M.M. King, and T.R. Soderling, Regulatory interactions of calmodulin-binding proteins: phosphorylation of calcineurin by autophosphorylated Ca2+/calmodulin-dependent protein kinase II. Proc Natl Acad Sci U S A, 1988. 85(18): p. 7001-5.

[92] Lee, C.W., et al., Interaction between salt-inducible kinase 2 and protein phosphatase 2A regulates the activity of calcium/calmodulin-dependent protein kinase I and protein phosphatase methylesterase-1. J Biol Chem, 2014. 289(30): p. 21108-19.

[93] Chiang, K.C., et al., MART-10, a less calcemic vitamin D analog, is more potent than 1alpha,25-dihydroxyvitamin D3 in inhibiting the metastatic potential of MCF-7 breast cancer cells in vitro. J Steroid Biochem Mol Biol, 2014. 139: p. 54-60.

[94] Chiang, K.C., et al., MART-10, the new brand of 1alpha,25(OH)2D3 analog, is a potent anti-angiogenic agent in vivo and in vitro. J Steroid Biochem Mol Biol, 2016. 155(Pt A): p. 26-34.

[95] Buitrago, C., M. Costabel, and R. Boland, PKC and PTPalpha participate in Src activation by 1alpha,25OH2 vitamin D3 in C2C12 skeletal muscle cells. Mol Cell Endocrinol, 2011. 339(1-2): p. 81-9.

[96] Bernichtein, S., et al., Vitamin D3 Prevents Calcium-Induced Progression of Early-Stage Prostate Tumors by Counteracting TRPC6 and Calcium Sensing Receptor Upregulation. Cancer Res, 2017. 77(2): p. 355-365.

[97] Berridge, M.J., Vitamin D cell signalling in health and disease. Biochem Biophys Res Commun, 2015. 460(1): p. 53-71.

[98] Christakos, S., et al., Vitamin D: beyond bone. Ann N Y Acad Sci, 2013. 1287: p. 45-58.

[99] Bandera Merchan, B., et al., The role of vitamin D and VDR in carcinogenesis: Through epidemiology and basic sciences. J Steroid Biochem Mol Biol, 2017. 167: p. 203-218.

[100] LaPorta, E. and J. Welsh, Modeling vitamin D actions in triple negative/basal-like breast cancer. J Steroid Biochem Mol Biol, 2014. 144 Pt A: p. 65-73.

[101] Welsh, J., Vitamin D and breast cancer: insights from animal models. Am J Clin Nutr, 2004. 80(6 Suppl): p. 1721S-4S.