Hormone dependence of breast cancer cells and the effects of tamoxifen and estrogen:31P NMR studies

Abstract

Many breast tumors appear to progress from estrogen-dependent growth to a more malignant phenotype characterized by estrogen-independent growth, antiestrogen resistance, and a high metastatic potential. Utilizing31P NMR spectroscopy on human breast cancer cells growingin vitro, we have investigated the effects of 17β-estradiol and tamoxifen on the metabolic/bioenergetic spectra of a series of human breast cancer cells that vary in their estrogen and antiestrogen responsiveness. A comparison of baseline spectra associates higher levels of phosphodiesters and UDP-glucosides (e.g. UDP-glucose, UDP-N-acetylglucosamine), and lower phosphocholine/glycerylphosphocholine and phosphocholine/phosphoethanolamine ratios, with the acquisition of estrogen-independent growth in estrogen receptor expressing cells. No metabolic changes are clearly associated with the metastatic phenotype. Whilst estrogen treatment produces no consistently significant spectral changes in any of the cell lines, the estrogen-independent and estrogen-responsive MCF7/MIII cell line responds to tamoxifen treatment by significantly increasing all spectral resonances 30%-40% above baseline values. This may reflect a tamoxifen-induced change to a more differentiated or apoptotic phenotype, or an attempt by the cells to reverse the inhibitory effects of the drug. The ability to detect metabolic changes in response to tamoxifen by NMR spectroscopy may provide a novel means to identify those tumors that are responsive to antiestrogen therapy.

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Abbreviations

CCS-IMEM:

steroid-deprived Improved Minimal Essential Medium

E2:

17β-estradiol

ER:

estrogen receptor

Pi :

inorganic phosphate

GPE:

glyceryl-phosphoethanolamine

GPC:

glyceryl-phosphocholine

PC:

phosphocholine

PE:

phosphoethanolamine

PDE:

phosphodiesters

PME:

phosphomonoesters

TAM:

tamoxifen (trans-1-(4-β-dimethylaminoethoxyphenyl)-1,2-diphenylbut-1-ene)

UDPG:

uridine diphosphoglycoside

References

  1. 1.

    Clarke R, Dickson RB, Brünner N: The process of malignant progression in human breast cancer. Ann Oncol 1: 401–407, 1990

    Google Scholar 

  2. 2.

    Leonessa F, Boulay V, Wright A, Thompson EW, Brünner N, Clarke R: The biology of breast tumor progression: acquisition of hormone-independence and resistance to cytotoxic drugs. Acta Oncol 31: 115–123, 1991

    Google Scholar 

  3. 3.

    King RJB, Stewart JF, Millis RR, Rubens RD, Hayward JL: Quantitative comparison of estradiol and progesterone receptor contents of primary and metastatic human breast tumors in relation to response to endocrine treatment. Breast Cancer Res Treat 2: 339–346, 1982

    Google Scholar 

  4. 4.

    Bronzert DA, Greene GL, Lippman ME: Selection and characterization of a breast cancer cell line resistant to the antiestrogen LY 117018. Endocrinology 117: 1409–1417, 1985

    Google Scholar 

  5. 5.

    Clarke R, Brünner N, Thompson EW, Glanz P, Katz D, Dickson RB, Lippman ME: The inter-relationships between ovarian-independent growth, antiestrogen resistance and invasiveness in the malignant progression of human breast cancer. J Endocrinol 122: 331–340, 1989

    Google Scholar 

  6. 6.

    Clarke R, Brünner N, Katzenellenbogen BS, Thompson EW, Norman MJ, Koppi C, Paik S, Lippman ME, Dickson RB: Progression from hormone dependent to hormone independent growth in MCF-7 human breast cancer cells. Proc Natl Acad Sci USA 86: 3649–3653, 1989

    Google Scholar 

  7. 7.

    Yano T, Korkut E, Pinski J, Szepeshazi K, Milovanovic S, Groot K, Clarke R, Comaru-Schally AM, Schally AV: Inhibition of growth of MCF-7 MIII human breast carcinoma in nude mice by treatment with agonists or antagonists of LHRH. Breast Cancer Res Treat 21: 35–45, 1992

    Google Scholar 

  8. 8.

    Brünner N, Frandsen TL, Holst-Hansen C, Bei M, Thompson EW, Wakeling AE, Lippman ME, Clarke R: MCF7/LCC2: A 4-hydroxytamoxifen resistant human breast cancer variant which retains sensitivity to the steroidal antiestrogen ICI 182, 780. Cancer Res 53: 3229–3232, 1993

    Google Scholar 

  9. 9.

    Coopman P, Garcia M, Brünner N, Derocq D, Clarke R, Rochefort H: Antiproliferative and antiestrogenic effects of ICI 164, 384 in 4-OH-tamoxifen-resistant human breast cancer cells. Int J Cancer 56: 295–300, 1994

    Google Scholar 

  10. 10.

    Thompson EW, Brünner N, Torri J, Johnson MD, Boulay V, Wright A, Lippman ME, Steeg PS, Clarke R: The invasive and metastatic properties of hormone-independent and hormone-responsive variants of MCF-7 human breast cancer cells. Clin Exp Metastasis 11: 15–26, 1993

    Google Scholar 

  11. 11.

    Brünner N, Boulay V, Fojo A, Freter C, Lippman ME, Clarke R: Acquisition of hormone-independent growth in MCF-7 cells is accompanied by increased expression of estrogen-regulated genes but without detectable DNA amplifications. Cancer Res 53: 283–290, 1993

    Google Scholar 

  12. 12.

    Clarke R, Thompson EW, Leonessa F, Lippman J, McGarvey M, Brünner N: Hormone resistance, invasiveness and metastatic potential in human breast cancer. Breast Cancer Res Treat 24: 227–239, 1993

    Google Scholar 

  13. 13.

    Merchant TE, Gierke LW, Meneses P, Glonek T:31P magnetic resonance spectroscopic profiles of neoplastic human breast tissues. Cancer Res 48: 5112–5118, 1988

    Google Scholar 

  14. 14.

    Smith TA, Glaholm J, Leach MO, Machin L, Collins DJ, Payne GS, McCready VR: A comparison ofin vitro andin vivo 31P NMR spectra from human breast tumors: variations in phospholipid metabolism. Br J Cancer 63: 514–516, 1991

    Google Scholar 

  15. 15.

    Neeman M, Degani H: Early estrogen-induced metabolic changes and their inhibition by actinomycin D and cycloheximide in human breast cancer cells:31P and13C NMR studies. Proc Natl Acad Sci USA 86: 5585–5589, 1989

    Google Scholar 

  16. 16.

    Kaplan O, Jaroszewski JW, Clarke R, Fairchild CR, Schoenlein P, Goldenberg S, Gottesman MM, Cohen JS: The multidrug resistance phenotype:31P NMR characterization and 2-deoxyglucose toxicity. Cancer Res 51: 1638–1644, 1991

    Google Scholar 

  17. 17.

    Kaplan O, Navon G, Lyon RC, Faustino PJ, Straka EJ, Cohen JS: Effects of 2-deoxyglucose on drug-sensitive and drug-resistant breast cancer cells: toxicity and magnetic resonance spectroscopy studies of metabolism. Cancer Res 50: 544–551, 1990

    Google Scholar 

  18. 18.

    Foxall DL, Cohen JS, Mitchell JB: Continuous perfusion of mammalian cells embedded in agarose gel threads. Exp Cell Res 154: 521–529, 1984

    Google Scholar 

  19. 19.

    Cohen JS, Lyon RC, Daly PF: Monitoring intracellular metabolism by NMR. Meth Enzymol 177: 435–438, 1989

    Google Scholar 

  20. 20.

    Strobl JS, Lippman ME: Prolonged retention of estradiol by human breast cancer cells in tissue culture. Cancer Res 39: 3319–3327, 1979

    Google Scholar 

  21. 21.

    Kaplan O, van Zijl PC, Cohen JS: Information from combined1H and31P NMR studies of cell extracts: differences in metabolism between drug-sensitive and drug-resistant MCF-7 human breast cancer cells. Biochem Biophys Res Commun 169: 383–390, 1990

    Google Scholar 

  22. 22.

    Shilabi Z, Dyer D: Protein analysis with bicinchoninic acid. Ann Clin Lab Sci 18: 235–239, 1988

    Google Scholar 

  23. 23.

    Clarke R, van den Berg HW, Murphy RF: Tamoxifen and 17β-estradiol reduce the membrane fluidity of human breast cancer cells. J Natl Cancer Inst 82: 1702–1705, 1990

    Google Scholar 

  24. 24.

    Bonomi PD, Finkelstein DM, Ruckdeschel JC, Blum RH: Combination chemotherapy versus single agents followed by combination chemotherapy in stage IV non-small cell lung cancer: a study of the Eastern Cooperative Oncology Group. J Clin Oncol 7: 1602–1613, 1989

    Google Scholar 

  25. 25.

    Corbett RJ, Nunnally RL, Giovanella BC, Antich PP: Characterization of the31P nuclear magnetic resonance spectra of human melanoma tumors implanted in nude mice. Cancer Res 47: 5065–5069, 1987

    Google Scholar 

  26. 26.

    Thomas M, Bader C, Monet JD: Sex steroid hormone modulation of NADPH pathways in MCF-7 cells. Cancer Res 50: 1195–1200, 1990

    Google Scholar 

  27. 27.

    Monet JD, Thomas M, Dautigny N, Brami M, Bader CA: Effects of 17 beta-estradiol and R5020 on glucose-6-phosphate dehydrogenase activity in MCF-7 human breast cancer cells: a cytochemical assay. Cancer Res 47: 5116–5119, 1987

    Google Scholar 

  28. 28.

    Neeman M, Degani H: Metabolic studies of estrogen and tamoxifen treated human breast cancer cells by nuclear magnetic resonance spectroscopy. Cancer Res 49: 589–594, 1989

    Google Scholar 

  29. 29.

    Stubbs M, Coombes RC, Griffiths JR, Maxwell RJ, Rodrigues LM, Gusterson BA:31P-NMR spectroscopy and histological studies of the response of rat mammary tumors to endocrine therapy. Br J Cancer 61: 258–262, 1990

    Google Scholar 

  30. 30.

    Smith TAD, Eccles S, Ormerod MG, Tombs AJ, Titley JC, Leach MO: The phosphocholine and glycerophosphocholine content of an oestrogen-sensitive rat mammary tumor correlates strongly with growth rate. Br J Cancer 64: 821–826, 1991

    Google Scholar 

  31. 31.

    Bindal RD, Carlson KE, Katzenellenbogen BS, Katzenellenbogen JA: Lipophilic impurities, not phenosulfonphthalein, account for the estrogenic properties in commerical preparations of phenol red. J Steroid Biochem 31: 287–293, 1988

    Google Scholar 

  32. 32.

    Sutherland RL, Reddel RR, Green MD: Effects of oestrogens on cell proliferation and cell cycle kinetics. A hypothesis on the cell cycle effects of antioestrogens. Eur J Cancer Clin Oncol 19: 307–318, 1983

    Google Scholar 

  33. 33.

    Merchant TE, Thelissen GRP, De Graaf PW, Den Otter W, Glonek T: Clinical magnetic resonance spectroscopy of human breast disease. Invest Radiol 26: 1053–1059, 1991

    Google Scholar 

  34. 34.

    Degani H, Horowitz A, Itzchak Y: Breast tumors: evaluation with P-31 NMR spectroscopy. Radiology 161: 53–55, 1986

    Google Scholar 

  35. 35.

    Price JE, Polyzos A, Zhang RD, Daniels LM: Tumorigenicity and metastasis of human breast carcinoma cell lines in nude mice. Cancer Res 50: 717–721, 1990

    Google Scholar 

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Correspondence to Robert Clarke.

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Ruiz-Cabello, J., Berghmans, K., Kaplan, O. et al. Hormone dependence of breast cancer cells and the effects of tamoxifen and estrogen:31P NMR studies. Breast Cancer Res Tr 33, 209–217 (1995). https://doi.org/10.1007/BF00665945

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Keywords

  • Estrogen
  • Tamoxifen
  • Human Breast Cancer Cell
  • High Metastatic Potential
  • Estrogen Receptor Express