Journal: Molecular Medicine Reports

Article Title: MicroRNA-328 directly targets p21-activated protein kinase 6 inhibiting prostate cancer proliferation and enhancing docetaxel sensitivity

doi: 10.3892/mmr.2015.4390

Figure Lengend Snippet: PAK6 expression in BPH and Pca tissue samples. (A) Pca samples were stained with anti-PAK6 (magnification, ×400). (B) BPH and (C–F) Pca tissue samples were stained with anti-PAK6 (magnification, ×200). Tissue samples with Gleason score of (C) <7; (D) 7 and (E) >7. (F) CRPC tissues. (G) PAK6 overexpression in CRPC samples was dectected by western blotting. PAK6, p21-activated protein kinase 6; BPH, benign prostate hyperplasia; Pca, prostate cancer; CRPC, castration-resistant Pca; ADPC, androgen-dependent Pca.

Article Snippet: A PAK6 3′-UTR-luciferase reporter was created by ligating the PAK6 3′-UTR PCR product into the Xho I and Not I restriction sites of the psiCHECK-2TM Vector (Promega Corp., Madison, WI, USA).

Techniques: Expressing, Staining, Over Expression, Western Blot

Journal: Molecular Medicine Reports

Article Title: MicroRNA-328 directly targets p21-activated protein kinase 6 inhibiting prostate cancer proliferation and enhancing docetaxel sensitivity

doi: 10.3892/mmr.2015.4390

Figure Lengend Snippet: PAK6 was directly targeted by miR-328. (A) miR-328 may bind to target sequences located in the nucleotides of the 3′-UTR of PAK6 mRNA (miRanda). (B) Quantitative reverse transcription-polymerase chain reaction identified decreased miR-328 expression levels in CRPC tissue samples compared with in ADPC tissues ( * P<0.05). (C) PAK6 protein was inhibited by treatment with miR-328 mimics, as evaluated by western blotting. (D) PAK6 3′-UTR was cloned into a luciferase reporter cloning site in the psiCHECK-2 dual luciferase vector. Luciferase activity markedly decreased with increasing miR-328 expression level ( * P<0.05). PAK6, p21-activated protein kinase 6; miR, microRNA; CRPC, castration-resistant prostate cancer; ADPC, androgen-dependent prostate cancer; NC, negative control; WT, wild-type; MUT, mutant.

Article Snippet: A PAK6 3′-UTR-luciferase reporter was created by ligating the PAK6 3′-UTR PCR product into the Xho I and Not I restriction sites of the psiCHECK-2TM Vector (Promega Corp., Madison, WI, USA).

Techniques: Reverse Transcription, Polymerase Chain Reaction, Expressing, Western Blot, Clone Assay, Luciferase, Cloning, Plasmid Preparation, Activity Assay, Negative Control, Mutagenesis