Summary
Lithium-stimulated MCF-7 cell proliferation was compared to proliferation stimulated by other mitogens for this cell line - estradiol (E2) and epidermal growth factor (EGF) - and lithium was found to be effective within a narrow concentration range. Mitogenic effects of lithium on proliferation stimulated by E2 and EGF were additive below maximum, but were not synergistic. The phosphoinositide pathway is a cell signaling system involved in cell proliferation, within which phospholipase C (PLC)-mediated hydrolysis of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] leads to the production of the second messengers inositol-1,4,5-trisphosphate [Ins(1,4,5)P3] and diacylglycerol (DAG), as well as to calcium mobilization. At mitogen concentrations which maximally stimulated cell growth, estradiol stimulated both growth and PLC activity, while EGF and lithium stimulated cell growth but had little effect on the activity of the enzyme. Dose-responses with EGF revealed that a low concentration (0.1 ng/ml, 0.017 nM) of EGF appeared to stimulate both PLC activity and cell growth, but that higher concentrations of EGF which stimulated greater proliferation inhibited PLC activity. Steady-state levels of inositol phosphates including inositol trisphosphate were increased by all three mitogens. In growth assays, the phorbol ester phorbol 12-myristate-13-acetate (PMA), which mimics the actions of DAG, stimulated some cell growth, but dioctanoylglycerol, an additional DAG analog, and the calcium ionophore A23187, alone or with the DAG analogs, had no effect. These results suggest that PLC-mediated PtdIns(4,5)P2 hydrolysis is not primarily associated with signaling proliferation by lithium or EGF in MCF-7 breast cancer cells.
References
Hori C, Oka T: Induction by lithium of multiplication of mouse mammary epithelium in culture. Proc Natl Acad Sci USA 76: 2823–2827, 1979
Ptashne K, Stockdale FE, Conlon S: Initiation of DNA synthesis in mammary epithelium and mammary tumors by lithium ions. J Cell Physiol 103: 41–46, 1980
Tomooka Y, Imagawa W, Nandi S, Bern HA: Growth effect of lithium on mouse mammary epithelial cells in serum-free collagen gel culture. J Cell Physiol 117: 290–296, 1983
Welshons WV, Grady LH, Judy BM: Control of proliferation of MCF-7 breast cancer cells in a commercial preparation of charcoal-stripped adult bovine serum. Breast Cancer Res Treat 23: 97–104, 1992
Freter CE, Lippman ME, Cheville A, Zinn S, Gelmann EP: Alterations in phosphoinositide metabolism associated with estradiol 17-β and growth factor treatment of MCF-7 breast cancer cells. Mol Endocrinol 2: 159–166, 1988
Etindi RN, Manni A, Martel MS: The effects of TGF-α and 17-β estradiol on phosphoinositide metabolism in MCF-7 breast cancer cells. Breast Cancer Res Treat 24: 61–70, 1992
Berridge MJ: Inositol trisphosphate and diacylglycerol: two interacting second messengers. Annu Rev Biochem 56: 159–193, 1987
Nishizuka Y: The role of protein kinase C in cell surface signal transduction and tumor promotion. Nature (Lond) 308: 693–698, 1984
Bell RM: Protein kinase C activation by protein kinase C second messengers. Cell 45: 631–632, 1986
Berridge MJ, Downes CP, Hanley MR: Neural and developmental actions of lithium: a unifying hypothesis. Cell 59: 411–419, 1989
Whitman M, Cantley L: Phosphoinositide metabolism and the control of cell proliferation. Biochem Biophys Acta 948: 327–344, 1988
Catt KJ, Balla T: Phosphoinositide metabolism and hormone action. Ann Rev Med 40: 487–509, 1989
Ignar-Trowbridge D, Hughes AR, Putney JW, McLachlan JA, Korach KS: Diethylstilbestrol stimulates persistent phosphatidylinositol lipid turnover by an estrogen receptor-mediated mechanism in immature mouse uterus. Endocrinology 129: 2423–2430, 1991
Bourke JE, Dank S, Wilce PA, Martin L: Effect of estradiol and progesterone on phosphatidylinositol metabolism in the uterine epithelium of the mouse. J Steroid Biochem 39: 337–342, 1991
Taylor MV, Metcalfe JC, Hesketh TR, Smith GA, Moore JP: Mitogens increase phosphorylation of phosphoinositides in thymocytes. Nature 312: 462–465, 1984
Wahl M, Nishibe S, Suh P-G, Rhee SG, Carpenter G: Epidermal growth factor stimulates tyrosine phosphorylation of phospholipase C-II independently of receptor internalization and extracellular calcium. Proc Natl Acad Sci (USA) 86: 1568–1572, 1989
Margolis B, Rhee SG, Felder S, Mervic M, Lyall R, Levitzki A, Ullrich A, Zilberstein A, Schlessinger J: EGF induces phosphorylation of phospholipase C-II: a potential mechanism for EGF receptor signaling. Cell 57: 1101–1107, 1989
Meisenhelder J, Suh P-G, Rhee SG, Hunter T: Phospholipase C-γ is a substrate for the PDGF and EGF receptor protein-tyrosine kinasesin vivo andin vitro. Cell 57: 1109–1122, 1989
Bollag G, McCormick F: Regulators and effectors ofras proteins. Ann Rev Cell Biol 7: 601–632, 1991
Hall A: Ras-related proteins. Current Opinion in Cell Biology 5: 265–268, 1993
Bone R, Springer JP, Atack JR: Structure of inositol monophosphatase, the putative target of lithium therapy. Proc Natl Acad Sci USA 89: 10031–10035, 1992
York JD, Majerus PW: Isolation and heterologous expression of a cDNA encoding bovine inositol polyphosphate 1-phosphatase. Proc Natl Acad Sci USA 87: 9548–9552, 1990
Schrey MP, Furlong MM, Holt JR, Patel KV: Heterologous regulation of inositol lipid hydrolysis in human breast cancer cells by oestradiol 17β, bombesin and fluoroaluminate. Int J Cancer 51: 93–98, 1992
Stack G, Gorski J: Estrogen-stimulated deoxyribonucleic acid synthesis: a ratchet model for the prereplicative period. Endocrinology 117: 2017–2023, 1985
Shechter Y, Hernaez L, Cuatrecasas P: Epidermal growth factor: biological activity requires persistent occupation of high-affinity cell surface receptors. Proc Natl Acad Sci USA 75: 5788–5791, 1978
Taylor JA, Welshons WV: MCF-7 breast cancer cell proliferation and membrane-associated phospholipase C activity: Implications for mitogenic signalling mechanisms (Abstract 1092). Endocrinology 130 (Suppl): 324, 1992
Read LD, Greene GL, Katzenellenbogen BS: Regulation of estrogen receptor messenger ribonucleic acid and protein levels in human breast cancer cell lines by sex steroid hormones, their antagonists, and growth factors. Mol Endocrinol 3: 295–304, 1989
West DC, Sattar A, Kumar S: A simplifiedin situ solubilization procedure for the determination of DNA and cell number in tissue cultured mammalian cells. Analyt Biochem 147: 289–295, 1985
Labarca C, Paigen K: A simple, rapid and sensitive DNA assay procedure. Analyt Biochem 102: 344–352, 1980
Bradford M: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt Biochem 72: 248–254, 1976
Jackowski S, Rettenmier CW, Sherr CJ, Rock CO: A guanine nucleotide-dependent phosphatidylinositol (4,5) diphosphate phospholipase C in cells transformed by the V-FMS or V-FES oncogenes. J Biol Chem 261: 4978–4985, 1986
Patel KV, Schrey MP: Activation of inositol phospholipid signaling and Ca2+ efflux in human breast cancer cells by bombesin. Cancer Res 50: 235–239, 1990
Batty IR, Nahorski SR, Irvine RF: Rapid formation of inositol 1,3,4,5-tetrakisphosphate following muscarinic receptor stimulation of rat cerebral cortical slices. Biochem J 232: 211–215, 1985
Castagna M, Takai Y, Kaibuchi K, Sano K, Kikkawa V, Nishizuka Y: Direct activation of calcium-activated, phospholipid dependent protein kinase by tumor promoting phorbol esters. J Biol Chem 257: 7847–7851, 1982
Issandou M, Bayard F, Darbon J-M: Inhibition of MCF-7 cell growth by 12-O-tetradecanoyl-13-acetate and 1,2-dioctanoyl-sn-glycerol: distinct effects on protein kinase C activity. Cancer Res 48: 6943–6950, 1988
Hart DA: Potentiation of phytohemagglutinin stimulation of lymphoid cells by lithium. Exp Cell Res 119: 47–53, 1979
Rybak SM, Stockdale FE: Growth effects of lithium chloride in BALB/c 3T3 fibroblasts and Madin-Darby canine kidney epithelial cells. Exp Cell Res 136: 263–270, 1981
Urabe M, Hershman JM, Pang X-P, Murakami S, Sugawara M: Effect of lithium on function and growth of thyroid cellsin vitro. Endocrinology 129: 807–814, 1991
Sjöholm Å, Welsh N, Hellerström C: Lithium increases DNA replication, polyamine content, and insulin secretion by rat pancreatic β-cells. Am J Physiol 262: C391-C395, 1992
Schou M: Pharmacology and toxicology of lithium. Annu Rev Pharmacol Toxicol 16: 231–243, 1976
Derynck R: The physiology of transforming growth factor-α. Adv Cancer Res 58: 27–52, 1992
Dickson RB, Huff KK, Spencer EM, Lippman ME: Induction of epidermal growth factor related polypeptides by 17-β estradiol in MCF-7 human breast cancer cells. Endocrinology 118: 138–142, 1986
Arteaga CL, Coronado E, Osborne CK: Blockade of the EGF receptor inhibits TGF-α induced but not estrogen-induced growth of hormone-dependent human breast cancer. Mol Endocrinol 2: 1064–1069, 1988
Cunningham TW, Kuppuswamy D, Pike LJ: Treatment of A431 cells with epidermal growth factor (EGF) induces desensitization of EGF-stimulated phosphatidylinositol turnover. J Biol Chem 264: 15351–15356, 1989
Nelson J, Connelly M, Walker B, Gray J, Shaw C, Murphy RF: Bombesin stimulates proliferation in human breast cancer cells in culture. Br J Cancer 63: 933–936, 1991
Zachary I, Miller J, Nanberg A, Higins T, Rozengurt E: Inhibition of bombesin-induced mitogenesis by pertussis toxin: dissociation from phospholipase C pathway. Biochem Biophys Res Comm 146: 456–463, 1987
Whitman M, Kaplan DR, Roberts TM, Cantley L: Evidence for two distinct phosphatidylinositol kinases in fibroblasts. Biochem J 247: 165–174, 1987
Kaplan D, Whitman M, Schaffhausen B, Palla DC, White M, Cantley L, Roberts D: Common elements in growth factor stimulation and oncogenic transformation: 85 kD phosphoprotein and phosphatidylinositol kinase activity. Cell 50: 1021–1029, 1987
Cohen B, Liu Y, Druker B, Roberts TM, Schaffhausen BS: Characterization of pp85, a target of oncogenes and growth factor receptors. Mol Cell Biol 10: 2909–2915, 1990
Auger KR, Serunian LA, Soltoff SP, Libby P, Cantley LC: PDGF-dependent tyrosine phosphorylation stimulates production of novel polyphosphoinositides in intact cells. Cell 57: 167–175, 1989
Ulug ET, Hawkins PT, Hanley MR, Courtneidge SA: Phosphatidylinositol metabolism in cells transformed by polyomavirus middle T antigen. J Virol 64: 3895–3904, 1990
Sylvia V, Curtin G, Norman J, Stec J, Busbee D: Activation of a low specific activity form of DNA polymerase α by inositol-1,4-bisphosphate. Cell 54: 651–658, 1988
Alvarez E, Northwood IC, Gonzalez FA, Latour DA, Seth A, Abate C, Curran T, Davis RJ: Pro-Leu-Ser/Thr-Pro is a consensus primary sequence for substrate protein phosphorylation. Characterization of the phosphorylation of c-myc and c-jun proteins by an epidermal growth factor receptor threonine 669 protein kinase. J Biol Chem 266: 15277–15285, 1991
Gille H, Sharrocks AD, Shaw PE: Phosphorylation of transcription factor p62TCF by MAP kinase stimulates ternary complex formation at c-fos promoter. Nature 358: 414–417, 1992
Pulverer BJ, Kyriakis JM, Avruch J, Nikolakaki E, Woodgett JR: Phosphorylation of c-jun mediated by MAP kinases. Nature 353: 670–674, 1991
Wu J, Dent P, Jelinek T, Wolfman A, Weber MJ, Sturgill TW: Inhibition of EGF-activated MAP kinase signaling pathway by adenosine 3',5'-monophosphate. Science 262: 1065–1069, 1993
Cook SJ, McCormick F: Inhibition by cAMP of Ras-dependent activation of Raf. Science 262: 1069–1072, 1993
Frödin M, Peraldi P, Van Obberghen E: Cyclic AMP activates the mitogen-activated protein kinase cascade in PC12 cells. J Biol Chem 269: 6207–6214, 1994
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Taylor, J.A., Grady, L.H., Engler, K.S. et al. Relationship of growth stimulated by lithium, estradiol, and EGF to phospholipase C activity in MCF-7 human breast cancer cells. Breast Cancer Res Tr 34, 265–277 (1995). https://doi.org/10.1007/BF00689718
Issue Date:
DOI: https://doi.org/10.1007/BF00689718