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Tumor Biology

Overexpression of bcl-2 Protects Prostate Cancer Cells from Apoptosis in Vitro and Confers Resistance to Androgen Depletion in Vivo

Anthony J. Raffo, Harris Perlman, Min-Wei Chen, Mark L. Day, Jack S. Streitman and Ralph Buttyan
Anthony J. Raffo
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Harris Perlman
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Min-Wei Chen
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Mark L. Day
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Jack S. Streitman
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Ralph Buttyan
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DOI:  Published October 1995
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Abstract

Normal (nonneoplastic) human prostatic secretory epithelial cells do not express the bcl-2 protein. However, a recent immunohistochemical survey of neoplastic human prostate tissues showed that a fraction of primary untreated prostate adenocarcinoma cells expressed this apoptosis-suppressing oncoprotein at significant levels (Colombel et al., Am. J. Pathol., 143: 390–400, 1993). Additionally, a number of hormone-refractory prostatic adenocarcinomas obtained from hormonally-treated patients (subsequent to surgical or drug castration therapy) were examined and were found to be uniform in their elevated expression of bcl-2 oncoprotein. The results of this preliminary survey imply that bcl-2 expression distinguishes a subgroup of primary human prostate cancers and that the expression of this protein might be a factor enabling prostate cancer cells to survive in an androgen-deprived environment. The current study was undertaken to determine the degree to which overexpression of bcl-2 can protect human prostate cancer cells from apoptotic stimuli in vitro and in vivo. Human prostate cancer cells (LNCaP) were transfected with a neomycin-selectable eucaryotic expression vector containing cDNA encoding human bcl-2. Transfected clonal variants that express bcl-2 protein (LNCaP/bcl-2) were unaltered with regard to their basal growth rate in 10% serum-containing medium, or with regard to their expression of the differentiated human prostate cell gene products prostate-specific antigen or androgen receptor protein. The bcl-2-transfected clones were altered, however, with regard to their growth rate in charcoal-stripped serum lacking dihydrotestosterone. Additionally, in contrast to the parental or control-transfected cell lines, LNCaP/bcl-2 cells were highly resistant to a variety of apoptotic stimuli in vitro including serum starvation and 10 nm phorbol ester (phorbol 12-myristate 13-acetate) supplementation of the medium. Lastly, the overexpression of bcl-2 by these prostate cancer cells altered their tumorigenic potential in a nude mouse assay. s.c. injections of 106 LNCaP/bcl-2 cells into male nude mice resulted in earlier and larger tumor formation compared to an equivalent injection of parental or control-transfected LNCaP cells. When these variant cell lines were injected into castrated male nude mice, only the LNCaP/bcl-2-transformed cells gave rise to tumors. Moreover, LNCaP/bcl-2 tumors grown in intact male nude mice were refractory to the growth-inhibiting effects of castration demonstrated by parental LNCaP cells. Data obtained in this study demonstrate that the bcl-2 oncoprotein can protect prostate cancer cells from apoptotic stimuli in vitro and suggest that such protection correlates with the ability to form hormone-refractory prostate tumors in vivo.

Footnotes

  • ↵1 This work was supported by United States Public Health Service Grants NIH-CA58089 and NIH-CA47848, as well as grants from the CaPCure Foundation and the Koch Foundation. M-W. C. is a research fellow of the American Foundation for Urological Disease.

  • ↵2 To whom requests for reprints should be addressed, at Department of Urology, Columbia University, College of Physicians and Surgeons, 630 West 168th Street, New York, NY 10032.

  • Received April 4, 1995.
  • Accepted August 2, 1995.
  • ©1995 American Association for Cancer Research.
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October 1995
Volume 55, Issue 19
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Overexpression of bcl-2 Protects Prostate Cancer Cells from Apoptosis in Vitro and Confers Resistance to Androgen Depletion in Vivo
Anthony J. Raffo, Harris Perlman, Min-Wei Chen, Mark L. Day, Jack S. Streitman and Ralph Buttyan
Cancer Res October 1 1995 (55) (19) 4438-4445;

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Overexpression of bcl-2 Protects Prostate Cancer Cells from Apoptosis in Vitro and Confers Resistance to Androgen Depletion in Vivo
Anthony J. Raffo, Harris Perlman, Min-Wei Chen, Mark L. Day, Jack S. Streitman and Ralph Buttyan
Cancer Res October 1 1995 (55) (19) 4438-4445;
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