caa a22 4500 397562047 CHVBK 20180308164818.0 cr unu---uuuuu 161202e19960703xx s 000 0 eng 10.1093/jnci/88.13.908 doi (NATIONALLICENCE)oxford-10.1093/jnci/88.13.908 Induction of Apoptosis by Diethylstilbestrol in Hormone-Insensitive Prostate Cancer Cells [Elektronische Daten] [Cary N. Robertson, Kathryn M. Roberson, George M. Padilla, E. Timothy O'Brien, J. Michael Cook, Choung-Soo Kim, Robert L. Fine] Background: Diethylstilbestrol (DES) and diethylstilbestrol diphosphate (DESdP) are effective agents for the treatment of advanced prostate cancers. Tumor-inhibiting effects of DES and DESdP are presumed secondary to suppression of androgen production in vivo. Little is known, however, about the direct cellular mechanisms of the tumor inhibition. Estrogens have been reported not only to stimulate growth but also to disrupt microtubule formation in prostate cancer cells. Purpose: The study was designed to examine and compare mechanisms of in vitro growth inhibition of DES and DESdP in human androgen-insensitive prostate cancer cells (DU145, 1-LN, and PC-3) and human androgen-sensitive prostate cancer cells (LNCaP) and to examine estrogen receptor modulation of such effects. Methods: The cytotoxic effects of DES and DESdP were examined in vitro by use of a standard microculture tetrazolium assay to quantitate numbers of viable cells. Immunofluorescence microscopy, DNA fragmentation analysis, and fluorescence flow cytometry were used to investigate microtubules, the induction of apoptosis, and changes in cell cycle distribution. The degree of estrogen receptor positivity of untreated and treated cells was determined by immunohistochemistry and quantitative image analysis. Results: LD50 levels (the dose at which 50% of cells are no longer viable) in the concentration range of 19-25 μM were observed for both DES and DESdP in all cell lines examined. DESdP-induced growth inhibition was found to be dependent on heat-labile phosphatases present in fetal calf serum. DES-induced cytotoxicity was not affected by the presence of 17β-estradiol, and it was not dependent on the presence of estrogen receptor. Estrogen receptor-positive cells and estrogen receptor-negative cells were equally responsive to DES. PC-3 cells stained with fluorescent anti-tubulin, phalloidin (actin stain), and 4', 6-diamidino-2-phenylindole (DNA stain) showed no inhibition of microtubules or actin filaments but revealed the presence of apoptotic bodies in the nuclei. Fluorescence flow cytometry of nuclear DNA content of propidium iodide-stained nuclei from androgen-insensitive prostate cancer cells treated with 15 or 30 μM DES or DESdP revealed an increase in relative numbers of hypodiploid (apoptotic) nuclei, a depletion of G1 and S-phase cells, and an accumulation of cells in G2/M phase. Conversely, androgen-sensitive cells contained a lower percentage of hypodiploid nuclei but no accumulation of cells in G2/M phase. Conclusions: Direct cytotoxic effects of DES in prostate cancer cells are estrogen receptor independent and do not involve disruption of microtubule architecture but do involve he promotion of cell cycle arrest and apoptosis. These are the first data confirming direct cytotoxic effects of DES and DESdP in prostate cancer cells via an apoptotic mechanism. Implications: These results suggest that DES and DESdP have potential value as agents against androgen-insensitive prostate neoplasms through induction of an apoptotic cascade. [J Natl Cancer Inst 1996; 88: 908-17] © Oxford University Press ARTICLES nationallicence Robertson Cary N. Department of Surgery, Division of Urology, Duke University and Durham VA Medical Centers NC aut Roberson Kathryn M. Department of Surgery, Division of Urology, Duke University and Durham VA Medical Centers NC aut Padilla George M. Department of Cell Biology, Duke University and Durham VA Medical Centers NC aut O'Brien E. Timothy Department of Ophthalmology, Duke University and Durham VA Medical Centers NC aut Cook J. Michael Comprehensive Cancer Center, Division of Flow Cytometry, Duke University and Durham VA Medical Centers NC aut Kim Choung-Soo Department of Surgery, Division of Urology, Duke University and Durham VA Medical Centers NC aut Fine Robert L. Division of Hematology-Oncology, Department of Medicine, Duke University and Durham VA Medical Centers NC aut JNCI: Journal of the National Cancer Institute Oxford University Press 88/13(1996-07-03), 908-917 0027-8874 88:13<908 1996 88 jnci https://doi.org/10.1093/jnci/88.13.908 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/jnci/88.13.908 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Robertson Cary N. Department of Surgery, Division of Urology, Duke University and Durham VA Medical Centers NC aut NATIONALLICENCE 700 1- Roberson Kathryn M. Department of Surgery, Division of Urology, Duke University and Durham VA Medical Centers NC aut NATIONALLICENCE 700 1- Padilla George M. Department of Cell Biology, Duke University and Durham VA Medical Centers NC aut NATIONALLICENCE 700 1- O'Brien E. Timothy Department of Ophthalmology, Duke University and Durham VA Medical Centers NC aut NATIONALLICENCE 700 1- Cook J. Michael Comprehensive Cancer Center, Division of Flow Cytometry, Duke University and Durham VA Medical Centers NC aut NATIONALLICENCE 700 1- Kim Choung-Soo Department of Surgery, Division of Urology, Duke University and Durham VA Medical Centers NC aut NATIONALLICENCE 700 1- Fine Robert L. Division of Hematology-Oncology, Department of Medicine, Duke University and Durham VA Medical Centers NC aut NATIONALLICENCE 773 0- JNCI: Journal of the National Cancer Institute Oxford University Press 88/13(1996-07-03), 908-917 0027-8874 88:13<908 1996 88 jnci Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford