Journal: Oncogenesis

Article Title: MicroRNA-18a is elevated in prostate cancer and promotes tumorigenesis through suppressing STK4 in vitro and in vivo

doi: 10.1038/oncsis.2014.12

Figure Lengend Snippet: miR-18a is highly expressed in tumors. ( a ) The expression level of miR-18a in prostate cancer tissue. The differential expression of miR-18a in prostate cancer tissues was compared with that in normal tissues. The level of miR-18a was significantly higher in cancer tissues ( P <0.01). ( b ) Left panel, ISH was used to detect the presence of miR-18a in non-tumor prostate and prostate cancer tissues. Representative ISH micrographs ( × 400) for miR-18a staining; right Panel, quantification according to ISH expression of miR-18a in prostate tumors as related to paired normal tissues. The scores are calculated as staining intensity × percentage of stained cells. ( c ) miR-18a relative expression level of non-tumor parts compared with tumor parts were analyzed using GEO data set. ( d ) The five NT paired tissues were used for examination of miR-18a expression. High expression of miR-18a tumor part specimens than that in non-tumor part tissues ( e ) The differential expression of miR-18a in prostate cancer cell lines was compared with that in prostate immortalized epithelium cell lines. The level of miR-18a was significantly higher in prostate cancer cell lines than in prostate immortalized epithelium cell lines.

Article Snippet: We used the prostate cancer tissue array obtained from BioMax Company (Rockville, MD, USA) to assay the correlation of miR-18a between TNM stage.

Techniques: Expressing, Quantitative Proteomics, Staining

Journal: Oncogenesis

Article Title: MicroRNA-18a is elevated in prostate cancer and promotes tumorigenesis through suppressing STK4 in vitro and in vivo

doi: 10.1038/oncsis.2014.12

Figure Lengend Snippet: STK4 is inversely correlated with miR-18a. ( a ) Venn diagram displaying miRNAs computationally predicted to target STK4 by PicTar (red), TargetScan (green), miRanda (blue) and Mirnatargets (black). Examination of different databases to find candidate downstream target genes. ( b ) Protein expression of STK4, SMAD2, CDC42, CTGF and α-tubulin in control miR or miR-18a precursor transfected prostate cancer cells (PC-3, DU145). STK4 was suppressed by miR-18a transfection at the protein level in prostate cancer cell lines as shown by western blotting. ( c ) The expression of STK4 mRNA (upper panel) and protein (bottom panel) in six prostate cell lines was analyzed by RT-PCR and western blotting, respectively. The number below the GAPDH image was the density ratio of STK4/GAPDH (NIH-Image J) for mRNA (upper panel) and protein (bottom panel), respectively. The STK4 mRNA level was the same in 60 cell lines. The STK4 protein level is downregulated in four prostate cancer cell lines. ( d ) Western blotting (upper panel) and IHC (middle panel) analysis revealed STK4 protein level was downregulated in prostate tumor parts compared with adjacent non-tumor tissues. Quantification of STK4 expression in paired prostate adenocarcinoma and corresponding normal tissues (bottom panel) ( n =10, P <0.001). Western blotting data showed that the STK4 protein is highly expressed in non-tumor compared with tumor tissues. IHC showed that STK4 staining is lost in the tumor tissues. ( e ) STK4 mRNA expression levels are not different between normal and tumor of prostate cancer in GEO data set validation. ( f ) ISH and IHC were used to detect the presence of miR-18a and STK4 in prostate cancer tissues. These sections of ISH and for IHC are consecutive sections. Quantitative IHC and ISH scores show that STK4 and miR-18a expression are inversely correlated. ( g ) miR-18a is highly expressed with poor tumor stage.

Article Snippet: We used the prostate cancer tissue array obtained from BioMax Company (Rockville, MD, USA) to assay the correlation of miR-18a between TNM stage.

Techniques: Expressing, Control, Transfection, Western Blot, Reverse Transcription Polymerase Chain Reaction, Staining, Biomarker Discovery

Journal: Oncogenesis

Article Title: MicroRNA-18a is elevated in prostate cancer and promotes tumorigenesis through suppressing STK4 in vitro and in vivo

doi: 10.1038/oncsis.2014.12

Figure Lengend Snippet: miR-18a increased cell survival by suppressing STK4-mediated AKT kinase activity to inhibit apoptosis. ( a ) MTT assay indicating that the overexpression of STK4 and antagomiR-18a induces cell death in 22Rv-1 cells. ( b ) Western blotting of miRNA-transfected cells to detect the PARP cleavage level. ( c ) AKT phosphorylation in prostate cancer cells was suppressed by ectopic STK4 expression. Our observation in LNcap and 22Rv-1 prostate cancer cells transfected with STK4 suppressed AKT phosphorylation activity and increased PARP cleavage protein. ( d ) Representative images show that the expression levels of STK4 are inversely related to the phosphorylation levels of AKT in prostate cancer xenograft tumor tissues. ( e ) Diagram indicating how miR-18a regulates STK4 to act as an oncomiR in prostate cancer.

Article Snippet: We used the prostate cancer tissue array obtained from BioMax Company (Rockville, MD, USA) to assay the correlation of miR-18a between TNM stage.

Techniques: Activity Assay, MTT Assay, Over Expression, Western Blot, Transfection, Phospho-proteomics, Expressing

Journal: Molecular cancer research : MCR

Article Title: AXL is a Putative Tumor Suppressor and Dormancy Regulator in Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-0718

Figure Lengend Snippet: AXL is not expressed in human prostate tumors. A, AXL expression prostate cancer patients who did and did not experience a biochemical recurrence (n = 1405 patients, GenomeDx GRID dataset). P value was calculated using the Wilcoxon rank test. B, Recurrence-free survival of patients with high (above the median) and low (below the median) AXL expression (n = 571 patients, Taylor et al, 2010 visualized and analyzed with Project Betastasis). C, Representative images of AXL IHC staining of a prostate tissue microarray; LN, lymph node. C42B cells served as a negative control, DU145 as a positive control. All images at original magnification x 200. D, AXL IHC staining in metastatic patient tissue; upper panel scale bar, 100 μm; lower panel scale bar, 50 μm.

Article Snippet: Human prostate tissue microarray (TMA) staining This study was approved by the Johns Hopkins institutional internal review board and followed the U.S. Common Rule.

Techniques: Expressing, Immunohistochemistry, Microarray, Negative Control, Positive Control