OBJECTIVE:The antiestrogen tamoxifen has been found to be effective in decreasing glioblas toma cell proliferation, but the mechanism underlying this effect and whether it is through the estrogen receptor (ER) is controversial. The objective of this study was to determine whether ERs are present in three human glioblastoma cell lines—HS683, U138MG, and JHN J889H—using the most sensitive techniques available.
METHODS: Ligand binding and flow cytometry were employed to identify estrogen and pro gesterone receptors. The reverse transcriptase-polymerase chain reaction was used to identify ER mRNA, and a novel reporter gene transfection assay demonstrated that the ER was capable of activating gene transcription.
RESULTS: U138MG glioblastoma cells contain ERs that are capable of increasing gene tran scription in response to estradiol. No ERs were found in HS683 or JHN J889H cells.
CONCLUSION: Tamoxifen may be acting through the ER in some glioblastoma cells. (J Soc Gynecol Invest 1994;1:238-44)
Poisson M., Pertuiset BF, Hauw J-J., et al. Steroid hormone receptors in human meningiomas, gliomas and brain metastases. J Neurooncol1983;1:179-89.
2.
Brentani MM, Lopes MT, Martins VR, Plese JPSteroid receptors in intracranial tumors. Clin Neuropharmacol1984;7:347-50.
3.
Fujimoto M., Yoshino E., Hirakawa K., Fujimoto J., Tamaya T.Estrogen receptors in brain tumors. Clin Neuropharmacol1984 ;7:357-62.
4.
Whittle IR, Hawkins RA, Miller JDSex hormone receptors in intracranial tumours and normal brain. Eur J Surg Oncol1987;13:303-7.
5.
Romic Stojković R, Jovančević M., Santel DU, Grčević N., Gamulin S.Sex steroid receptors in intracranial tumors. Cancer1990;65:1968-70.
6.
Pollack IF, Randall MS, Kristofik MP, Kelly RH, Selker RG, Vertosick FTEffect of tamoxifen on DNA synthesis and proliferation of human malignant glioma lines in vitro. Cancer Res1990;50:7134-8.
7.
Vertosick FT , Selker RG, Pollack IF, Arena V.The treatment of intracranial malignant gliomas using orally administered tamoxifen therapy: Preliminary results in a series of "failed" patients. Neurosurgery1992;30:897-903.
8.
Su HD, Mazzei GJ, Vogler WR, Kuo JFEffects of tamoxifen, a nonsteroidal antiestrogen, on phospholipid/calciumdependent protein kinase and phosphorylation of its endogenous substrate proteins from the rat brain and ovary. Biochem Pharmacol1985;34:3649-53.
9.
Owens RB, Smith HS, Nelson-Rees WA, Springer ELEpithelial cell cultures from normal and cancerous human tissues. J Natl Cancer Inst1976;56:843-9.
10.
Ponten J., Macintyre EHLong term culture of normal and neoplastic human glia. Acta Pathol Microbiol Scand1968; 74:465-86.
11.
Beckman G., Beckman L., Ponten J., Westermark B.G-6-PD and PGM phenotypes of 16 continuous human tumor cell lines. Evidence against cross-contamination by HeLa cells. Hum Hered1971;21:238-41.
12.
Tanaka K., Testa JRAssessment of methods for the cytogenetic analysis of human solid tumors. J Natl Cancer Inst1987;79:1287-93.
13.
Chirgwin JM , Przybyla AE, MacDonald RJ, Rutter WJIsolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry1979;18: 5294-9.
14.
Green S., Walter P., Kumar V., et al. Human oestrogen receptor cDNA: Sequence, expression and homology to v-erb-A. Nature1986;320:134-9.
15.
Ponglikitmongkol M., Green S., Chambon P.Genomic organization of the human oestrogen receptor gene. EMBO J1988;7:3385-8.
16.
Southern EMDetection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol1975;98:503-17.
17.
Feherty P., Farrer-Brown G., Kellie AEOestradiol receptors in carcinoma and benign disease of the breast: An in vitro assay. Br J Cancer1971;25:697-710.
18.
Horwitz KB, McGuire WLProgesterone and progesterone receptors in experimental breast cancer . Cancer Res1977; 37:1733-8.
19.
Burton K.A study of the conditions and mechanism of diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J1956;62:315.
20.
Graham ML II, Krett NL, Miller LA, et al. T47Dco cells, genetically unstable and containing estrogen receptor mutations, are a model for the progression of breast cancers to hormone resistance . Cancer Res1992;50:6208-17.
21.
Graham ML II, Smith JA, Jewett PB, Horwitz KBHeterogeneity of progesterone receptor content and remodeling by tamoxifen characterize subpopulations of cultured human breast cancer cells: Analysis of quantitative dual parameter flow cytometry. Cancer Res1992;52:593-602.
22.
Graham ML II, Bunn PA Jr, Jewett PB, Gonzalez-Aller C., Horwitz KBSimultaneous measurement of progesterone receptors and DNA indices by flow cytometry: Characterization of an assay in breast cancer cell lines . Cancer Res1989;49:3934-42.
23.
Klein-Hitpass L., Schorpp M., Wagner U., Ryffel GUAn estrogen-responsive element derived from the 5' flanking region of the Xenopus vitellogenin A2 gene functions in transfected human cells. Cell1986;46:1053-61.
24.
Leslie KK, Tasset D., Horwitz KBFunctional analysis of a mutant estrogen receptor isolated from T47Dco breast cancer cells. Am J Obstet Gynecol1992 ;166:1053-61.
25.
Vegeto E., Cocciolo MG, Raspagliesi F., Piffanelli A., Fontanelli R., Maggi A.Regulation of progesterone receptor gene expression. Cancer Res1990;50:5291-5.
26.
Eckert RL, Katzenellenbogen BSEffects of estrogens and antiestrogens on estrogen receptor dynamics and the induction of progesterone receptor in MCF-7 human breast cancer cells . Cancer Res1982;42:139-44.
27.
Nardulli AM , Greene GJ, O'Malley BW, Katzenellenbogen BSRegulation of progesterone receptor messenger ribonucleic acid and protein levels in MCF-7 cells by estradiol: Analysis of estrogen effects on progesterone synthesis and degradation. Endocrinology1988;122:935-44.
28.
Horwitz KB, Mockus MB, Lessey BAVariant T47D human breast cancer cells with high progesterone receptor levels despite estrogen and antiestrogen resistance. Cell1982;28:633-42.
29.
Nishida M., Kasahara K., Kaneko M., Iwasaki H.Establishment of a new human endometrial adenocarcinoma cell line, Ishikawa cells, containing estrogen and progesterone receptors. Nippon Sanka Fujinka Gakkai Zasshi1985;37: 1103-11.
30.
Holinka CF, Hata H., Kuramoto H., Gurpide E.Responses to estradiol in a human endometrial adenocarcinoma cell line (Ishikawa) . J Steroid Biochem1986;24:85-9.
31.
Coezy E., Borgna J-L., Rochefort H.Tamoxifen and metabolites in MCF7 cells: Correlation between binding to estrogen receptor and inhibition of cell growth. Cancer Res1982;42:317-23.
32.
Cho H., Katzenellenbogen BSSynergistic activation of estrogen receptor-mediated transcription by estradiol and protein kinase activators. Mol Endocrinol1993;7:441-52.
33.
Dickson RB, Lippman MEEstrogenic regulation of growth and polypeptide growth factor secretion in human breast carcinoma. Endocr Rev1987 ;8:29-43.
34.
Lippman ME, Dickson RB, Gelmann EP, et al. Mechanisms of estrogenic and antiestrogenic regulation of growth of human breast carcinoma. In: Klijin JGM , ed. Hormonal manipulation of cancer: Peptides, growth factors, and new (anti)steroidal agents. New York: Raven Press, 1987:381-403.
