Lithium suppresses cell proliferation by interrupting E2F–DNA interaction and subsequently reducing S–phase gene expression in prostate cancer

Abstract

These data indicated that LiCl suppresses cancer cell proliferation by disrupting E2F-DNA interaction and subsequent E2F-mediated gene expression in prostate cancer.

Serum/glucocorticoid-induced protein kinase-1 facilitates androgen receptor-dependent cell survival

“…After stimulation with R1881 (1.0 nM) or the solvent in 2% cFBS-containing media, cells were subjected to ChIP assay, as described. 38 Anti-AR immunoprecipitated chromatin DNA were amplified with three different primer sets, and 10% of whole cellular genomic DNA were used as PCR templates as the positive control. Data represent two different experiments Figure 4a, transfection with the SGK-1 siRNA but not the control siRNA resulted in a dramatic loss of SGK-1 expression.…”

“…The human prostate cancer LNCaP and LAPC-4 cells and their culture conditions were described previously. 12,33,[38][39][40] LAPC-4/ SGK1 and LNCaP/SGK1 sublines were established by stably infecting the parental cells with a retrovirus vector (pMSCV-puro-SGK1) 29 bearing the rat sgk1 gene. The control subline LAPC-4/Puro and LNCaP/Puro were established when an empty vector was used.…”

“…Genechip s arrays (Affymetrix Inc., Santa Clara, CA) was utilized and the protocol was described in our recent publication. 38 Briefly, 10 mg of total RNAs were used for the first-strand cDNA synthesis with the SuperscriptChoice system (Invitrogen). The in vitro-transcription labeling was conducted using the EnzoBio array high-yield RNA transcript labeling kit (Affymetrix).…”

“…For protein expression analysis, western blot was performed as described previously. 12,33,[38][39][40] Briefly, after treatment, cells were pelleted and lysed in a buffer containing protease inhibitors (Halft Protease Inhibitor Cocktail Kit, PIERCE, Rockford, IL), separated on SDS-PAGE gels and transferred to PVDF membrane (BIO-RAD, Hercules, CA). Membranes were blocked in a Tris-buffered solution plus 0.1% Tween-20 (TBS-T) solution with 5% nonfat dry milk and incubated with primary antibodies overnight at 41C.…”

“…The ChIP assay was performed using a ChIP-ITt assay kit (Active Motif, Carlsbad, CA) as described in our recent publication. 38 Cells were maintained in 15-cm dishes and serum-starved for 24 h. After pretreatment with anti-androgen Bicalutamide (Casodex, CSDX), cells were stimulated with R1881. Polyclonal antibodies against AR (Clone C815), RNA polymerase II, GAPDH, and normal rabbit serum were obtained from Santa Cruz Biotech (Santa Cruze, CA).…”