Figure 1

MREs of miR-1, miR-133 and miR-218 enabled adenovirus-mediated adenoviral vector to express TRAIL with bladder cancer specificity. (a) Illustration was shown of the structures of the involved adenoviral vectors. Ad-TRAIL-MRE-1-133-218 contained MREs of miR-1, miR-133 and miR-218 that were inserted immediately following TRAIL gene. ITR: inverse terminal region. (b) qPCR assay was performed to detect TRAIL mRNA expression. TRAIL mRNA levels in Ad-TRAIL cells were selected as standards and GAPDH was selected as endogenous reference. Means ± SEM of three independent experiments were shown. (c) TRAIL protein level was also determined in T24 and RT-4 bladder cells as well as BMCs infected with different adenoviruses by immunoblotting. GAPDH was selected as endogenous reference. (d) TRAIL protein level was also evaluated in the same cells infected with the indicated adenoviruses by ELISA assay. Means ± SEM of three independent experiments were shown. (e) T24 cells were treated with both 10 MOI of Ad-TRAIL-MRE-1-133-218 and mixed mimics of miR-1, miR-133 and miR-218 (100 nM for each) or control mimics (300 nM). 48 h later, TRAIL expression was tested by immunoblotting assay. GAPDH was selected as endogenous reference.