Journal: Cancer Reports
Article Title: Retinoic acid receptor γ is a therapeutically targetable driver of growth and survival in prostate cancer
doi: 10.1002/cnr2.1284
Figure Lengend Snippet: RARγ promotes growth and inhibits differentiation of prostate cancer cell lines. (A‐C) CV‐1 cells were transiently transfected with expression vectors encoding a fusion protein containing the estrogen receptor (ER) DNA binding domain (DBD) and the ligand binding domain (LBD) of the indicated RAR isotype, together with a firefly luciferase reporter under the control of an estrogen response element (ERE‐Tk‐ Luc ). Differences in transfection efficiency between samples were normalized using a control vector constitutively expressing Renilla luciferase. Transfected cells were treated with (A) ATRA, (B) AGN195183, or (C) AGN205327 at the indicated concentrations for 24 hours analyzed for luciferase activity. Results shown are from a typical experiment performed in triplicate and are expressed as relative light units (RLU, arbitrary units) ± SD that have been normalized to the maximal response to ATRA (10 −6 M). The compound concentrations at which half‐maximal effect is observed (EC 50 ) for RARα, RARβ, or RARγ are indicated. (D) Serum‐free adapted LNCaP cells were treated with the indicated concentrations of pan‐RAR agonist ATRA, RARγ‐selective agonist AGN205327, RARα‐selective agonists AGN195183 and AM580. Results are presented as the mean number of cells recovered (±SD). (E) Serum‐free adapted LNCaP cells were treated for 4 days with ATRA (10 −8 M), AGN195183 (10 −8 M) and the RARα‐selective antagonist AGN196996 (10 −8 M) as indicated. Results are presented as the mean number of cells recovered (±SD). (F) Serum‐free adapted LNCaP, PC‐3, or DU145 cells were grown in differentiation medium without compound (Control), containing AGN205327 (10 −7 M) or PPARγ‐selective antagonist GW9662 (10 −5 M) as indicated. 14 days post‐induction of differentiation, cells were tested for lipid droplet accumulation using the Oil red O assay. Results are presented as mean fold increase in absorbance at 510 nm (±SD) relative to undifferentiated serum‐free adapted cells (Untreated). (G) Adipogenic differentiation of serum‐free adapted LNCaP cells was induced by treatment with PPARγ‐selective agonist ciglitazone (10 −6 M). AGN205327 (10 −7 M) was added at the indicted times pre‐ or post‐treatment with ciglitazone. Cells were analyzed and results presented as described in (F). Statistical significance is represented as described in Section
Article Snippet: All‐ trans ‐retinoic acid (Cat. # 0695); the pan‐RAR agonist TTNPB (4‐[( E )‐2‐(5,6,7,8‐Tetrahydro‐5,5,8,8‐tetramethyl‐2‐naphthalenyl)‐1‐propenyl]benzoic acid, Cat. # 0761) ; the RARα‐selective agonist AM580 (4‐[(5,6,7,8‐Tetrahydro‐5,5,8,8‐tetramethyl‐2‐naphthalenyl)carboxamido]benzoic acid, Cat. # 0760) ; the PPARγ‐selective agonist ciglitazone (Cat. # 1307) ; the PPARγ‐selective irreversible antagonist GW9662 (Cat. # 1508) ; the microtubule stabilizer docetaxel (Cat. # 4056); the thymidylate synthetase inhibitor 5‐fluorouracil (Cat. # 3257); and the topoisomerase II inhibitor etoposide (Cat. # 1226) were purchased from Tocris Biosciences.
Techniques: Transfection, Expressing, Binding Assay, Ligand Binding Assay, Luciferase, Plasmid Preparation, Activity Assay
