Skip to main content

Advertisement

SpringerLink
Log in

Induction of apoptosis by AN-152, a cytotoxic analog of luteinizing hormone-releasing hormone (LHRH), in LHRH-R positive human breast cancer cells is independent of multidrug resistance-1 (MDR-1) system

  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Objective. More than 50% of human breast cancers express receptors for luteinizing hormone-releasing hormone (LHRH-R). These receptors can be used for targeted chemotherapy with agents like AN-152, in which doxorubicin is linked to analog [D-Lys<sup>6</sup>]LHRH. We compared the effects of AN-152 and doxorubicin in human breast cancer cells.

Methods. MCF-7, T47D, HCC-70 and ZR-75-1 cells were analysed for expression of LHRH-R using RT-PCR, immunostaining and flow cytometry. Apoptosis and expression of MDR-1 gene product Pgp were measured by flow cytometry. Cleavage of doxorubicin from AN-152 by serum carboxylesterase (CE) was inhibited by DFP.

Results. In MCF-7, T47D and HCC-70 cells we found cell surface expression of LHRH-R. In ZR-75-1 cells only sparse surface expression was found. In HCC-70 cells, induction of apoptosis by AN-152 was stronger than by doxorubicin in medium containing fetal calf serum (FCS). Pretreatment with DFP increased AN-152-induced apoptosis in LHRH-R positive MCF-7 and HCC-70 cells and reduced apoptosis in ZR-75-1 cells. In serum-free medium apoptosis induced by AN-152 was stronger than that induced by doxorubicin in HCC-70, T47D and MCF-7 cells, but weaker in ZR-75-1 cells. In medium containing FCS, both AN-152 and doxorubicin induced surface expression of MDR-1 gene product Pgp, but the effect of AN-152 was weaker. Pgp-expression induced by AN-152 was inhibited by pretreatment with DFP. AN-152 did not induce Pgp-expression in serum-free medium.

Conclusion.The cytotoxic LHRH analog AN-152 induces apoptosis independent of MDR-1 in LHRH-R positive breast cancer cells. The efficacy and/or specificity of AN-152 is improved by suppression or absence of CE activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Fekete M, Wittli JL, Schally AV:Characteristics and distribution of receptors for [D-TRP6]-luteinizing hormone-releasing hormone,somatostatin,epidermal growth factor, and sex steroids in 500 biopsy samples of human breast cancer.J Clin Lab Anal 3:137–147,1989

    Google Scholar 

  2. Baumann KH, Kiesel L, Kaufmann M, Bastert G, Runne-baum B:Characterization of binding sites for a GnRH-agonist (buserelin)in human breast cancer biopsies and their distribution in relation to tumor parameters.Breast Cancer Res Treat 25:37–46,1993

    Google Scholar 

  3. Moriya T, Suzuki T, Pilichowska M, Ariga N, Kimura N, Ouchi N, Nagura H, Sasano H:Immunohistochemical expression of gonadotropin releasing hormone receptor in human breast carcinoma.Pathol Int 51:333–337,2001

    Google Scholar 

  4. Eidne KA, Flanagan CA, Harris NS, Millar RP:Gonado-tropin releasing hormone (GnRH)-binding sites in human breast cancer cell lines and inhibitory effects of GnRH-agonists.J Clin Endocrinol Metab 64:425–432,1987

    Google Scholar 

  5. Mullen P, Scott WN and Miller WR:Growth inhibition observed following administration of an LHRH agonist to a clonal variant of the MCF-7 breast cancer cell line is accompanied by an accumulation of the cells in the G0 /G1 phase of the cell cycle.Br J Cancer 63:930–932,1991

    Google Scholar 

  6. Klijn JG:LH-RH agonists in the treatment of metastatic breast cancer:ten years' experience.Recent Results Cancer Res 124:75–90,1992

    Google Scholar 

  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 LH-RH.Breast Cancer Res Treat 21:35–45, 1992

    Google Scholar 

  8. Marini L, Iacopino F, Schinzari G, Robustelli della Cuna FS, Mantovani G, Sica G:Direct antiproliferative effect of triptorelin on human breast cancer cells.Anticancer Res 14: 1881–1885,1994

    Google Scholar 

  9. Segal-Abramson T, Kitroser H, Levy J, Schally AV, Sharoni Y:Direct effects of luteinizing hormone-releasing hormone agonists and antagonists on MCF-7 mammary cancer cells.Proc Natl Acad Sci USA 89:2336–2339, 1992

    Google Scholar 

  10. Vo ¨lker P, Gru ¨ndker C, Schmidt O, Schulz KD, Emons G: Expression of receptors for luteinizing hormone-releasing hormone in human ovarian and endometrial cancers: frequency,autoregulation,and correlation with direct antiproliferative activity of luteinizing hormone-releasing hormone analogues.Am J Obstet Gynecol 186:171–179, 2002

    Google Scholar 

  11. Verschraegen CF, Westphalen S, Hu W, Loyer E, Kudelka A, Volker P, Kavanagh J, Steger M, Schulz KD, Emons G: Phase II study of cetrorelix,a luteinizing hormone-releasing hormone antagonist in patients with platinum-resistant ovarian cancer.Gynecol Oncol 90:552–559,2003

    Google Scholar 

  12. Schally AV, Nagy A:New approaches to treatment of various cancers based on cytotoxic analogs of LHRH, somatostatin and bombesin.Life Sci 72:2305–2320,2002

    Google Scholar 

  13. Emons G, Ortmann O, Schulz KD, Schally AV:Growth-inhibitory actions of luteinizing hormone-releasing hor-mone on tumor cells.Trends Endocrinol Metab 8:355–362, 1997

    Google Scholar 

  14. Schally AV:Luteinizing hormone-releasing hormone ana-logs;their impact on the control of tumorigenesis.Peptides 20:1247–1262,1999

    Google Scholar 

  15. Krebs LJ, Wang X, Pudavar HE, Bergey EJ, Schally AV, Nagy A, Prasad PN, Liebow C:Regulation of targeted chemotherapy with cytotoxic lutenizing hormone-releasing hormone analogue by epidermal growth factor.Cancer Res 60:4194–4199,2000

    Google Scholar 

  16. Halmos G, Nagy A, Lamharzi N, Schally AV:Cytotoxic analogs of luteinizing hormone-releasing hormone bind with high a.nity to human breast cancers.Cancer Lett 136: 129–136,1999

    Google Scholar 

  17. Wang X, Krebs LJ, Al-Nuri M, Pudavar HE, Ghosal S, Liebow C, Nagy AA, Schally AV, Prasad PN:A chemically labeled cytotoxic agent:two-photon.uorophore for optical tracking of cellular pathway in chemotherapy.Proc Natl Acad Sci USA 96:11081–11084,1999

    Google Scholar 

  18. Szepeshazi K, Schally AV, Nagy A, Halmos G, Groot K: Targeted cytotoxic luteinizing hormone-releasing hormone (LHRH)analogs inhibit growth of estrogen independent MXT mouse mammary cancers in vivo by decreasing cell proliferation and inducing apoptosis.Anticancer Drugs 8: 974–987,1997

    Google Scholar 

  19. Baja AM, Schally AV, Halmos G, Nagy A:Targeted doxorubicin-containing luteinizing hormone-releasing hor-mone analogue AN-152 inhibits the growth of doxorubicin-resistant MX-1 human breast cancers.Clin Cancer Res 9: 3742–3748,2003

    Google Scholar 

  20. Early Breast Cancer Trialists' Collaborative Group:Poly-chemotherapy for early breast cancer:an overview of the randomised trials.The Lancet 352:930–942,1998

    Google Scholar 

  21. Booser DJ, Hortobagyi GN:Anthracycline antibiotics in cancer therapy:focus on drug resistance.Drugs 47:223–258,1994

    Google Scholar 

  22. Zhang W, Ling V:Cell-cycle-dependent turnover of P-glycoprotein in multidrug-resistant cells.J Cell Physiol 184:17–26,2000

    Google Scholar 

  23. Nagy A, Plonowski A, Schally AV:Stability of cytotoxic luteinizing hormone-releasing hormone conjugate (AN-152) containing doxorubicin 14-O-hemiglutarate in mouse and human serum in vitro:implications for the design of preclinical studies.Proc Natl Acad Sci USA 97:829–834, 2000

    Google Scholar 

  24. Emons G, Ortmann O, Becker M, Irmer G, Springer B, Laun R et al.:High a.nity binding and direct antiprolif-erative effects of LHRH analogues in human ovarian cancer cell lines.Cancer Res 53:5439–5446,1993

    Google Scholar 

  25. Emons G, Schro¨der B, Ortmann O, Westphalen S, Schulz K-D, Schally AV:High a.nity binding and direct antipro-liferative e.ects of luteinizing hormone-releasing hormone analogs in human endometrial cancer cell lines.J Clin Endocrinol Metab 77:1458–1464,1993

    Google Scholar 

  26. Vo¨lker P, Gru¨ndker C, Schmidt O, Schulz KD, Emons G: Expression of receptors for luteinizing hormone-releasing hormone in human ovarian and endometrial cancers: frequency,autoregulation and correlation with direct anti-proliferativer activity of LHRH analogues.Am J Obstet Gynecol 186:171–179,2002

    Google Scholar 

  27. Nicoletti I, Migloriati G, Pagliacci MC, Grignani F, Ricardi C:A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and.ow cytometry.J Immunol Meth 139:271–279,1991

    Google Scholar 

  28. Nagy A, Schally AV, Armatis P, Szepeshazy K, Halmos G, Kovacs M et al.:Cytotoxic analogs of luteinizing hormone-releasing hormone containing doxorubicin or 2-pyrollino-doxorubicin,a derivate 500–1000 times more potent.Proc Natl Acad Sci USA 93:7269–7273,1996

    Google Scholar 

  29. Chatzistamou I, Schally AV, Nagy A, Armatis P, Szepes-hazi K, Halmos G:Effective treatment of metastatic MDA-MB-435 human estrogen-independent breast carcinomas with a targeted cytotoxic analogue of luteinizing hormone-releasing hormone AN-207.Clin Cancer Res 6:4158–4165, 2000

    Google Scholar 

  30. Westphalen S, Kotulla G, Kaiser F, Krauß W, Werning G, Elsa¨sser HP et al.:Receptor mediated antiproliferative effects of the cytotoxic LHRH agonist AN-152 in human ovarian and endometrial cancer cell lines.Int J Oncol 17: 1063–1069,2000

    Google Scholar 

  31. Miyazaki M, Nagy A, Schally AV, Lamharzi N, Halmos G, Szepeshazi K et al.:Growth inhibition of human ovarian cancers by cytotoxic analogues of luteinizing hormone-releasing hormone.J Natl Cancer Inst 89:1803–1809,1997

    Google Scholar 

  32. Arencibia JM, Schally AV, Krupa M, Bajo AM, Nagy A, Szepeshazi K, Plonowski A:Targeting of doxorubicin bES-2 human ovarian cancers in nude mice by linking to an analog of luteinizing hormone-releasing hormone improves its e.ectiveness.Int J Oncol 19:571–577,2001

    Google Scholar 

  33. Gru¨ndker C, Vo¨lker P, Griesinger F, Ramaswamy A, Nagy A, Schally AV, Emons G:Antitumor effects of the cytotoxic luteinizing hormone-releasing hormone analog AN-152 on human endometrial and ovarian cancers xenografted into nude mice.Am J Obstet Gynecol 187:528–537, 2002

    Google Scholar 

  34. Letsch M, Schally AV, Szepeshazy K, Halmos G, Nagy A: Preclinical evaluation of targeted cytotoxic luteinizing hormone-releasing hormone analogue AN-152 in andro-gen-sensitive and insensitive prostate cancers.Clin Cancer Res 9:4505–4513,2003

    Google Scholar 

  35. Mechetner E, Kyshtoobayeva A, Zonis S, Kim H, Stroup R, Garcia R, Parker RJ, Fruehauf JP:Levels of multi-drug resistance (MDR1)P-glycoprotein expression by human breast cancer correlate with in vitro resistance to taxol and doxorubicin.Clin Cancer Res 4:389–398, 1998

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Gynecology and Obstetrics, Robert-Koch-Street 40, Germany

    Andreas R. Gumlnthert, Carsten Grumlndker, Till Bongertz & Gumlnter Emons

  2. Tulane University Medical Center and Veterans Affairs Medical Center, LA, USA

    Attila Nagy & Andrew V. Schally

Authors
  1. Andreas R. Gumlnthert
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Carsten Grumlndker
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Till Bongertz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Attila Nagy
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Andrew V. Schally
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gumlnter Emons
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gumlnthert, A.R., Grumlndker, C., Bongertz, T. et al. Induction of apoptosis by AN-152, a cytotoxic analog of luteinizing hormone-releasing hormone (LHRH), in LHRH-R positive human breast cancer cells is independent of multidrug resistance-1 (MDR-1) system. Breast Cancer Res Treat 87, 255–264 (2004). https://doi.org/10.1007/s10549-004-8806-8

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-004-8806-8

Search

Navigation