Reduction of wild type p53 function confers a hormone resistant phenotype on LNCaP prostate cancer cells.
BACKGROUND
The protein encoded by the p53 gene is required for some forms of apoptosis and loss or mutations in this gene are found with increased frequency in advanced and hormone resistant human prostate cancers. In order to better appreciate whether reduction of wildtype p53 function in prostate cancer cells might contribute to the development of therapeutic-resistance by these cells, we created stable variants of the androgen-responsive, wild type p53-expressing human prostate cancer cell line, LNCaP, by transfection with expression vectors designed to reduce expression or function of wildtype p53 in them. These cells were then tested for their ability to form tumors in castrated male nude mice.


METHODS
A conditional eukaryotic expression vector (under tetracycline regulation) expressing antisense p53 cDNA was constructed and either directly transfected into LNCaP cells or tranduced into these cells using recombinant retroviruses containing the vector. Stably transfected/transduced cells (LNCaP/Asp53) were evaluated by Western blot analysis for the ability of doxycycline to reduce p53 protein expression and for their ability to form tumors in castrated male nude mice treated or untreated with doxycycline. Additionally, we derived an LNCaP subline (LNCaP/DD) stably expressing a dominant-negative form of p53 and tested these cells for their ability to form tumors in castrated male nude mice.


RESULTS
LNCaP/Asp53 cells showed reduced expression of p53 protein when cultured in a medium containing doxycycline and tested sublines were able to efficiently form tumors in castrated male nude mice only when the mice were treated with doxycycline. LNCaP/DD cells were readily able to form tumors in castrated male nude mice whereas parental LNCaP cells or control-transfected LNCaP cells were not.


CONCLUSIONS
Loss of wildtype p53 function can contribute to the phenotype of hormone resistance of prostate cancer cells.
Burchardt
M
M
The Department of Urology, The College of Physicians and Surgeons of Columbia University, New York, USA.
Burchardt
T
T
Shabsigh
A
A
Ghafar
M
M
Chen
M W
MW
Anastasiadis
A
A
de la Taille
A
A
Kiss
A
A
Buttyan
R
R
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015153:Blotting, Western; D016373:DNA, Antisense / Q000235:genetics / Q000494:pharmacology; D004318:Doxycycline / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009376:Neoplasms, Hormone-Dependent / Q000235:genetics* / Q000473:pathology; D009919:Orchiectomy; D017354:Point Mutation; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000473:pathology; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
D000900:Anti-Bacterial Agents; D016373:DNA, Antisense; D016159:Tumor Suppressor Protein p53; D004318:Doxycycline