NF-κB-Activated lncRNACASC9 Promotes Bladder Cancer Progression by Regulating the TK1 Expression

Figure 6

CASC9 directly interacts with miR-195-5p. (a) Relative CASC9 expression levels in the nuclear and cytoplasm fractions of 5637 and T24 cells. Nuclear controls: U6, cytosolic controls: GAPDH. (b) FISH was used to detect CASC9 localization in 5637 and T24 cells. Red: CASC9; blue: DAPI. (c) RNA pull-down assays were used to examine the association of CASC9 and potential target microRNAs. (d) The MiR-195-5p expression was examined in 5637 cells transfected with CASC9 siRNAs (left) or overexpression vectors (right). (e) The MiR-195-5p expression was analyzed by qRT-PCR in BC and adjacent nontumor tissues (). (f) Luciferase reporter assays were used to determine the interacting activity between miR-195-5p and CASC9. (f) RIP assays with an anti-Ago2 antibody to assess endogenous Ago2 binding RNAs and IgG was used as the negative control. The levels of CASC9 and miR-195-5p were determined by qRT–PCR and presented as fold enrichment in Ago2 relative to input. (g) The correlation between CASC9 and miR-195-5p was analyzed in 30 paired BC samples (, ). , , and .

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