Journal: Cell Death & Disease

Article Title: Downregulation of miR-205 and miR-31 confers resistance to chemotherapy-induced apoptosis in prostate cancer cells

doi: 10.1038/cddis.2010.85

Figure Lengend Snippet: miR-205 targets Bcl-w and miR-31 targets E2F6. ( a ) Upper panel, predicted duplex formation between human BCL2L2 3′UTR and miR-205. Lower panel, predicted duplex formation between human E2F6 3′UTR and miR-31. ( b ) Left, WPE1-NB26 cells were transfected with GFP- BCL2L2 ( 3′UTR) or GFP- BCL2L2 (3′UTR)-mut plasmids together with plasmids expressing miR-205 or a negative control miRNA. Western blotting was performed with anti-GFP and anti-actin antibodies. Right, WPE1-NB26 cells were transfected with GFP- E2F6 ( 3′UTR) or GFP- E2F6 (3′UTR)-mut plasmids together with plasmids expressing miR-31 or a negative control miRNA. Western blotting was performed with anti-GFP and anti-actin antibodies. ( c ) Proteins were isolated from the indicated cell lines, western blots were done using indicated antibodies. ( d ) Left, WPE1-NB26 cells were transfected with pcDNA6.2-GW/EmGFP-miR-negative control or pcDNA6.2-GW/EmGFP-miR205, or pcDNA6.2-GW/EmGFP-miR31 plasmids. Expression of miRNAs was confirmed by real-time PCR. At 48 h after transfection, cell lysates were analyzed by western blots with the indicated antibodies (Middle). Right, WPE1-NA22 cells were transfected with synthetic miRNA inhibitors specific to miR-205 or miR-31, or the negative control inhibitor. At 48 h after transfection, cell lysates were analyzed by western blots with the indicated antibodies. Representative images of three independent experiments were shown

Article Snippet: To determine whether differential miRNA expression has a role in the apoptosis resistance in WPE1-NB26 cells, we compared miRNA expression profiles between WPE1-NA22 and WPE1-NB26 cells, using the mir Vana miRNA Bioarray.

Techniques: Transfection, Expressing, Negative Control, Western Blot, Isolation, Real-time Polymerase Chain Reaction

Journal: Cell Death & Disease

Article Title: Downregulation of miR-205 and miR-31 confers resistance to chemotherapy-induced apoptosis in prostate cancer cells

doi: 10.1038/cddis.2010.85

Figure Lengend Snippet: miR-205 and miR-31 sensitize WPE1-NB26 cells to chemotherapy-induced apoptosis. ( a ) WPE1-NB26 cell line stably expressing miR-205 was established and miR-205 expression was confirmed by real-time PCR. Negative control miRNA or miR-205 expressing cells were treated with various doses of Docetaxel for 24 h, and apoptosis was measured by Cell Death Detection Elisa analysis as described in Materials and Methods section. ( b ) Negative control or miR-205 expressing WPE1-NB26 cells were treated with various doses of Cisplatin for 24 h, and apoptosis was detected as above. ( c ) WPE1-NB26 cell line stably expressing miR-31 was established and miR-31 expression was confirmed by real-time PCR. Negative control miRNA or miR-31 expressing cells were treated with various doses of Docetaxel for 24 h, and apoptosis was measured as above. ( d ) Negative control miRNA or miR-31 expressing WPE1-NB26 cells were treated with various doses of Cisplatin for 24 h, and apoptosis was detected as above. The experiments have been repeated three times, data shown are mean values+S.D. ( * P <0.05, *** P <0.001)

Article Snippet: To determine whether differential miRNA expression has a role in the apoptosis resistance in WPE1-NB26 cells, we compared miRNA expression profiles between WPE1-NA22 and WPE1-NB26 cells, using the mir Vana miRNA Bioarray.

Techniques: Stable Transfection, Expressing, Real-time Polymerase Chain Reaction, Negative Control, Enzyme-linked Immunosorbent Assay