The GnRH agonist Goserelin acetate [D-Ser(tBu)6Aza-Gly10-GnRH, Zoladex, GnRH-A] was kindly provided by AstraZeneca Pharmaceuticals. The GnRH antagonist Antide (Ant) and docetaxel (Doc) were purchased from Sigma-Aldrich.
In all the experiments, the GnRH agonist has been utilized at the dose of 10−6 mol/L, on the basis of previous studies, from the authors’ laboratory as well as from others, aimed to investigate the molecular aspects of the antitumor activity of GnRH analogs in CRPC cells [29] (link), [41] (link)–[46] (link). The same range of doses were also utilized to investigate the antitumor activity of GnRH agonists in several experimental models of cancer cells overexpressing the GnRH receptor [16] (link), [47] (link)–[49] (link).
Pifithrin-α, the specific inhibitor of p53 transcriptional activity, and SB203580, the specific p38 MAPK inhibitor, were purchased from Santa Cruz Biotechnology and from Sigma-Aldrich, respectively.
Antibodies used for Western blotting experiments: rabbit anti-human Bax (1∶500; #2772), rabbit anti-human p38 MAPK (1∶1,000;. #9212) and rabbit anti-human p-p38 MAPK (1∶1,000; #9211) from Cell Signaling Technology; mouse monoclonal anti-human Bcl-2 (1∶250; #Sc-7382), mouse monoclonal anti-human p53 (1∶1,000; #Sc-53394), rabbit anti-human p-p53 (Ser-15; 1∶1,000; #Sc-101762) and goat anti-human actin 1–19 (1∶1,000; #Sc-1616) from Santa Cruz Biotechnology.
In all the experiments, the GnRH agonist has been utilized at the dose of 10−6 mol/L, on the basis of previous studies, from the authors’ laboratory as well as from others, aimed to investigate the molecular aspects of the antitumor activity of GnRH analogs in CRPC cells [29] (link), [41] (link)–[46] (link). The same range of doses were also utilized to investigate the antitumor activity of GnRH agonists in several experimental models of cancer cells overexpressing the GnRH receptor [16] (link), [47] (link)–[49] (link).
Pifithrin-α, the specific inhibitor of p53 transcriptional activity, and SB203580, the specific p38 MAPK inhibitor, were purchased from Santa Cruz Biotechnology and from Sigma-Aldrich, respectively.
Antibodies used for Western blotting experiments: rabbit anti-human Bax (1∶500; #2772), rabbit anti-human p38 MAPK (1∶1,000;. #9212) and rabbit anti-human p-p38 MAPK (1∶1,000; #9211) from Cell Signaling Technology; mouse monoclonal anti-human Bcl-2 (1∶250; #Sc-7382), mouse monoclonal anti-human p53 (1∶1,000; #Sc-53394), rabbit anti-human p-p53 (Ser-15; 1∶1,000; #Sc-101762) and goat anti-human actin 1–19 (1∶1,000; #Sc-1616) from Santa Cruz Biotechnology.