Mifepristone-induced secretion of transforming growth factor beta1-induced apoptosis in prostate cancer cells.
Successful therapy should induce apoptosis in prostate cancer cells irrespective of their androgen response. We have investigated the possibility of utilizing Mifepristone and Tamoxifen as treatment options for prostate cancer cells. Because preliminary results demonstrated induction of apoptosis by these drugs, the mechanism of induction of apoptosis was investigated. LNCaP-C4 prostate cancer cells were treated with Mifepristone and/or Tamoxifen. To confirm cytotoxic effects, nude mice with LNCaP-C4 xenografts were treated with Mifepristone and Tamoxifen. Cell viability was assayed using Sulforhodamine B (SRB) assay and DNA fragmentation was measured by ELISA. Culture media from vehicle- and drug-treated cells were collected and secretion of transforming growth factor beta1 (TGFbeta1) was estimated by ELISA. Role of TGFbeta1 was confirmed by inhibiting its function using TGFbeta1 antibody or M6P, which blocked activation of TGFbeta1. Apoptotic effects were determined by immunoblots of cytochrome c levels in cytosol and by in vitro colorimetric assay of caspase-3 activity. Results showed that although both drugs induced apoptosis in LNCaP-C4 cells, Mifepristone was more effective. The effects of these drugs on xenografts confirmed in vitro results. It was hypothesized that drug-induced secretion of TGFbeta1 may be responsible for induction of apoptosis. Neutralization of TGFbeta1 with an antibody or blocking the activation of TGFbeta1 by M6P abrogated the effects of Mifepristone and Tamoxifen confirming our hypothesis. Furthermore, treatment with Mifepristone and/or Tamoxifen released cytochrome c into the cytoplasm and induced activity of caspase-3, providing evidence that the drug-stimulated secretion of TGFbeta1 was responsible for induction of apoptosis in these cells. In conclusion, both Mifepristone and Tamoxifen induced apoptosis mediated through TGFbeta1. However, no critical advantage was noted by the addition of Tamoxifen to Mifepristone treatment.
Liang
Yayun
Y
Medical College of Georgia, Section of Urology, Augusta, GA 30912-4050, USA.
Eid
Manal A
MA
El Etreby
Fathy
F
Lewis
Ronald W
RW
Kumar
M Vijay
MV
D000818:Animals; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D003574:Cytochrome c Group / Q000378:metabolism; D003600:Cytosol; D004797:Enzyme-Linked Immunosorbent Assay; D004965:Estrogen Antagonists / Q000494:pharmacology*; D005456:Fluorescent Dyes; D006801:Humans; D015151:Immunoblotting; D008297:Male; D051379:Mice; D008819:Mice, Nude; D015735:Mifepristone / Q000494:pharmacology*; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology*; D012235:Rhodamines; D013629:Tamoxifen / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D053773:Transforming Growth Factor beta1; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured / Q000187:drug effects
D003574:Cytochrome c Group; D004965:Estrogen Antagonists; D005456:Fluorescent Dyes; D012235:Rhodamines; C508960:TGFB1 protein, human; C508961:Tgfb1 protein, mouse; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D013629:Tamoxifen; C022027:lissamine rhodamine B; D015735:Mifepristone; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases