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Drug-induced Apoptosis by Anti-microtubule Agent, Estramustine Phosphate on Human Malignant Glioma Cell Line, U87MG; in vitro Study

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Abstract

The drug effect of estramustine phosphate (EMP), an anti-microtubule agent on human glioma cells has been studied with the focus being mainly its cytotoxity or its targeting of organelles. However, the pharmacological knowledge of estramustine with respect to its cytotoxity and mechanism is limited. To acquire such knowledge, the present study investigates the ability of EMP to induce apoptosis in a human malignant glioma cell line. Transmission electron microscope (TEM) images were examined to monitor periodic changes. Agarose gel electrophoresis was also examined. Cellular DNA fragmentation ELISA was performed to investigate the DNA fragmentation rates and an MTT assay was studied to evaluate the ID50. A TEM study revealed condensing and fragmentation of the chromatin. Laddering of the bands was observed in all EMP exposure groups in agarose gel electrophoresis. DNA fragmentation in all EMP groups began at 0.5 h following an exposure with EMP and increased in a dose- and time-dependent manner as revealed by DNA ELISA fragmentation. ID50 at 24 h was 5.0 µM according to the MTT assay, a value close to 4.8 µM of ID50 was revealed by the DNA fragmentation assay. None of the above mentioned changes was observed in the control group. These results indicated that EMP caused a drug-induced apoptosis in the human malignant glioma cell line, U87MG.

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References

  1. Sorger P, Dobles M, Tounebize R, Hyman A: Coupling cell division and cell death to microtubule dynamics. Curr Opin Cell Biol 9: 07-814, 1997

    Google Scholar 

  2. Yoshida D, Piepmeier JM, Teramoto A: In vitro inhibition of cell proliferation, viability, and invasiveness in U87MG human glioblastoma cells by estramustine phosphate. Neurosurgery 39: 360-366, 1996

    Google Scholar 

  3. Pittman S, Geyp M, Fraser M, Ellem K, Peaston A, Ireland A: Multiple centrosomal microtubule organising centres and inversed microtubule stability are early features of VP-16-induced apoptosis in CCRF-CEM cells. Leukem Res 21: 491-499, 1997

    Google Scholar 

  4. Weller M, Trepel M, Grimmel C, et al.: Hypericin-induced apoptosis of human malignant glioma cells is light-dependent, independent of bcl-2 expression, and does not require wild-type p53. Neurol Res 19: 459-470, 1997

    Google Scholar 

  5. Fisher D: Apoptosis in cancer therapy. Cell 78: 539-542, 1994

    Google Scholar 

  6. Jordan M, Toso R, Throewr D, Wison L: Mechanism of mitotic block and inhibition of cell proliferation by taxol at low concentrations. Proc Natl Acad Sci USA 90: 9552-9556, 1993

    Google Scholar 

  7. Jordan M, Wendell K, Gardner S, Derry W, Copp H, Wilson L: Mitotic block induced in HeLa cells by low concentrations of peclitaxol (Taxol) results in abnormal mitotic exit and apototic cell death. Cancer Res 56: 846-825, 1996

    Google Scholar 

  8. Lin CH, Forscher P: Cytoskeleton remodeling during growth cone-target interactions. J Cell Biol 121: 1369-1383, 1993

    Google Scholar 

  9. Woods C, Zhu J, MCQueney P, Bollag D, Lazarides E: Taxol-induced mitotic block triggers rapid onset of a p53-independent apoptotic pathway. Mol Med 1: 506-526, 1995

    Google Scholar 

  10. Vallbo C, Bergenheim A, Bergh A, Grankvist K, Henriksson R: DNA fragmentation induced by the antimitotic drug estramustine in malignant rat glioma but not in normal brain-suggesting an apoptotic cell death. Br J Cancer 71: 717-720, 1995

    Google Scholar 

  11. Bergenheim A, Henriksson R, Piepmeier J, Yoshida D: Estramustine in malignant glioma. J Neuro-Oncol 30: 81-89, 1996

    Google Scholar 

  12. Piepmeier J, Pedersen P, Yoshida D, Greer C: Targeting microtubule-associated proteins in glioblastoma: A new strategy for selective therapy. Ann Surg Oncol 3: 543-549, 1996

    Google Scholar 

  13. Bergenheim AT, Bjork P, Bergh J, von-Schoultz E, Svedberg H, Henriksson R: Estramustine-binding and specific binding protein of the anti-mitotic compound estramustine in astrocytoma. Cancer Res 54: 4974-4979, 1994

    Google Scholar 

  14. Forsgren B, Björk P: Specific binding of estramustine to prostatic proteins. J Urol 23: 31-38, 1984

    Google Scholar 

  15. Moraga D, Rivas-Berrios A, Farías G, Wallin M, Maccioni RB: Estramustine-phosphate binds to a tubulin binding domain on microtubule-associated proteins MAP-2 and tau. Biochem Biophys Acta 1121: 97-103, 1992

    Google Scholar 

  16. Sheridan VR, Speicher LA, Tew KD: The effects of estramustine on metaphase and anaphase in DU 145 prostatic carcinoma cells. Eur J Cell Biol 54: 268-276, 1991

    Google Scholar 

  17. Hartley-Asp B: Estramustine-induced mitotic arrest in two human prostatic carcinoma cell lines DU 145 and PC-3. The Prostate 5: 93-100, 1984

    Google Scholar 

  18. Kim JH, Khil MS, Ryu S, Gabel M: Clinical and biological studies of estramustine phosphate as a radiation sensitizer. Int J Radiat Oncol Biol Phys 29: 555-557, 1994

    Google Scholar 

  19. Ryu S, Gabel M, Khil MS, Lee YJ, Kim SH, Kim JH: Estramustine: A novel radiation enhancer in human carcinoma cells. Int J Radiat Oncol Biol Phys 30: 99-104, 1994

    Google Scholar 

  20. Yoshida D, Piepmeier JM, Weinstein M: Estramustine sensitizes human glioblastoma cells to irradiation. Cancer Res 54: 1415-1417, 1994

    Google Scholar 

  21. Yoshida D, Peipmeier J, Bergenheim A, Henriksson R, Teramoto A: Suppression of matrix metalloproteinase-2-mediated cell invasion in U87MG, human glioma cells by anti-microtubule agent: In vitro study. Br J Cancer 77: 21-25, 1998

    Google Scholar 

  22. Johansson M, Bergenheim A, D'Argy R, et al.: Distribution of estramustine in the BT4C rat glioma model. Cancer Chemother Pharmacol 41: 317-325, 1998

    Google Scholar 

  23. Hudes GR, Greenberg R, Krigel RL, et al.: Phase 2 study of Estramustine and Vinblastine, two microtubule inhibitors, in hormone-refractory prostate cancer. J Clin Oncol 10: 1754-1761, 1992

    Google Scholar 

  24. Burton K: A study of the conditions and mechanisms of the diphenylalamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J 62: 315-323, 1956

    Google Scholar 

  25. Gunnarsson PO, Andersson SB, Sandberg AA, Ellman M: Accumulation of estramustine and estromustine in adipose tissue of rats and humans. Cancer Chemother Pharmacol 28: 361-364, 1991

    Google Scholar 

  26. von-Shoultz E, Grankvist K, Gustavson H, Henriksson R: Effects of estramustine on DNA and cell membrane in malignant glioma cells. Acta Oncol 30: 719-723, 1991

    Google Scholar 

  27. Piepmeier JM, Keefe DL, Weinstein MA, et al.: Estramustine and estrone analogs rapidly and reversibly inhibit deoxyribonucleic acid synthesis and alter morphology in cultured human glioblastoma cells. Neurosurgery 32: 422-431, 1993

    Google Scholar 

  28. Bergenheim AT, Gunnarsson PO, Edman K, Schoultz EV, Hariz M: Uptake and retension of estramustine and presence of estramustine binding protein in malignant brain tumours in humans. Br J Cancer 67: 358-361, 1993

    Google Scholar 

  29. Rutberg M, Wallin M: Estramustine induces disorganization of microtubules, perinuclear retraction of vimentin and endoplasmatic reticulum, and inhibits cell migration. Acta Histochem 95: 155-167, 1993

    Google Scholar 

  30. Dahllof B, Billstrom A, Cabral F, Hartley-Asp B: Estramustine depolymerizes microtubules by binding to tubulin. Cancer Res 53: 4573-4581, 1993

    Google Scholar 

  31. Engstrom KG, Grankvist K, Henriksson R: Early morphological detection of estramustine cytotoxity measured as alteration in cell size and shape by a new technique of microperifusion. Eur J Cancer 27: 1288-1295, 1991

    Google Scholar 

  32. Yoshida D, Cornell-Bell A, Piepmeier JM: Selective antimitotic effects of estramustine correlate with its antimicrotubule properties on glioblastoma and astrocytes. Neurosurgery 34: 863-368, 1994

    Google Scholar 

  33. Stearns ME, Wang M, Sousa O: Evidence that estramustine binds MAP-1 A to inhibit type IV collagenase secretion. J Cell Sci 98: 55-63, 1991

    Google Scholar 

  34. von-Shoultz E, Begenheim T, Grankvist K, Henriksson R: Estramustine binding protein in human brain-tumor tissue. J Neurosurg 74: 962-964, 1991

    Google Scholar 

  35. Reed J: Regulation by Bcl-2 family proteins and its role in cancer and chemoresistance. Curr Opin Oncol 7: 541-546, 1995

    Google Scholar 

  36. Stewart B: Mechanisms of apoptosis' integration of genetic, biochemical and cellular factors. J Natl Cancer Inst 86: 1285-1295, 1994

    Google Scholar 

  37. Rieder C, Schultz A, Cole R, Sluder G: Anaphase onset in vertebrate somatic cells is controlled by a checkpoint that monitors sister kinetochore attachment to the spindle. J Cell Biol 127: 1301-1310, 1994

    Google Scholar 

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Yoshida, D., Hoshino, S., Shimura, T. et al. Drug-induced Apoptosis by Anti-microtubule Agent, Estramustine Phosphate on Human Malignant Glioma Cell Line, U87MG; in vitro Study. J Neurooncol 47, 133–140 (2000). https://doi.org/10.1023/A:1006393705560

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