Development of a hammerhead ribozyme against BCL-2. II. Ribozyme treatment sensitizes hormone-resistant prostate cancer cells to apoptotic agents.
BACKGROUND
Several lines of evidence strongly implicate a crucial role for the apoptosis suppressing bcl-2 oncogene in the genesis of hormone-refractory human prostate cancer. By efficiently destroying the intracellular bcl-2 mRNA, one might be able to make the prostate cancer cell responsive again to conventional apoptotic stimuli such as androgen withdrawal. To achieve this end, we have devised a catalytic antisense RNA strategy (Ribozyme) for bcl-2 and evaluated its gene therapeutic potential.


RESULTS
Bcl-2 overexpressing LNCaP prostatic carcinoma cells (LNCaP/bcl-2) were transfected with the anti-bcl-2 ribozyme RNA using a polyamine-based transfection reagent and the reduction in the intracellular bcl-2 mRNA levels was followed by a ribonuclease protection assay. Using a cell viability assay, prior ribozyme transfection and subsequent application of apoptotic stimuli such as serum starvation or phorbol ester treatment caused a 30% increase in cell death by apoptosis than with these apoptotic stimuli alone.


CONCLUSIONS
The results obtained strongly support the ability of a potential anti-bcl-2 ribozyme therapy to synergize with other agents in inducing apoptosis of hormone-resistant human prostate cancer cells.
Dorai
T
T
Department of Urology, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
Goluboff
E T
ET
Olsson
C A
CA
Buttyan
R
R
D017209:Apoptosis / Q000187:drug effects; D004357:Drug Synergism; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006131:Growth Inhibitors / Q000008:administration & dosage; D006801:Humans; D008297:Male; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D016337:RNA, Catalytic / Q000008:administration & dosage / Q000737:chemistry* / Q000627:therapeutic use; D012367:RNA, Viral / Q000235:genetics; D014407:Tumor Cells, Cultured
D006131:Growth Inhibitors; D019253:Proto-Oncogene Proteins c-bcl-2; D016337:RNA, Catalytic; D012367:RNA, Viral