Abstract | PURPOSE: MATERIALS AND METHODS: Immunohistochemical analysis of prostate tissues, ELISA, and northern blotting of LNCaP cell lines were carried out for detection of tumor necrosis factor-alpha ( TNFalpha). To see the effect of TNFalpha on androgen receptor (AR), western blotting and northern blotting were performed after extraction of total protein and total RNA from LNCaP cells. Regulation of androgen-sensitivity by TNFalpha was investigated with cell proliferation assay and luciferase assay using PSA promoter after transfection of LNCaP cells. RESULTS: Immunohistochemical analysis demonstrated that TNFalpha protein was strongly expressed in epithelial cells of prostate cancer tissue but not in normal prostatic tissue. Basal level of TNFalpha in cell culture medium from LNCaP cells was very low. However, 12-O-tetradecanoylphorbol 13-acetate (TPA) induced TNFalpha secretion into medium up to 1600 pg/ml/day. Furthermore, 24 hr. post-TPA treatment TNFalpha mRNA levels were increased 15-fold compared to pre-treatment levels. TNFalpha (0 to 30 ng./ml. for 4 days) repressed AR protein and mRNA levels in a dose-dependent fashion in LNCaP cells. Pre-treatment of cells with actinomycin D treatment revealed that repression of mRNA levels was exerted at the post-transcriptional level. TNFalpha inhibited the ability of 10-9 M dihydrotestosterone (DHT) to induce LNCaP cell proliferation and activation of the prostate specific antigen (PSA) gene promoter. This inhibition was partially reversed by overexpression of transgenic androgen receptor. CONCLUSIONS:
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Authors | A Mizokami, A Gotoh, H Yamada, E T Keller, T Matsumoto |
Journal | The Journal of urology
(J Urol)
Vol. 164
Issue 3 Pt 1
Pg. 800-5
(Sep 2000)
ISSN: 0022-5347 [Print] United States |
PMID | 10953159
(Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
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Chemical References |
- Antibiotics, Antineoplastic
- Carcinogens
- RNA, Messenger
- RNA, Neoplasm
- Receptors, Androgen
- Tumor Necrosis Factor-alpha
- Dihydrotestosterone
- Dactinomycin
- Luciferases
- Prostate-Specific Antigen
- Tetradecanoylphorbol Acetate
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Topics |
- Antibiotics, Antineoplastic
(pharmacology)
- Blotting, Northern
- Blotting, Western
- Carcinogens
(pharmacology)
- Cell Division
(drug effects)
- Dactinomycin
(pharmacology)
- Dihydrotestosterone
(antagonists & inhibitors)
- Disease Progression
- Dose-Response Relationship, Drug
- Enzyme-Linked Immunosorbent Assay
- Gene Expression Regulation, Neoplastic
- Humans
- Immunohistochemistry
- Luciferases
(analysis)
- Male
- Promoter Regions, Genetic
(drug effects, genetics)
- Prostate-Specific Antigen
(genetics)
- Prostatic Neoplasms
(physiopathology)
- RNA Processing, Post-Transcriptional
(drug effects)
- RNA, Messenger
(drug effects)
- RNA, Neoplasm
(analysis, genetics)
- Receptors, Androgen
(drug effects, genetics)
- Tetradecanoylphorbol Acetate
(pharmacology)
- Tumor Cells, Cultured
- Tumor Necrosis Factor-alpha
(genetics, pharmacology)
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