Research Paper Volume 12, Issue 7 pp 6191—6205

Long non-coding RNA TPT1-AS1 promotes angiogenesis and metastasis of colorectal cancer through TPT1-AS1/NF90/VEGFA signaling pathway

Figure 5. TPT1-AS1 directly interacts with NF90, which enhances the stability of VEGFA mRNA. (A) The potential binding site that was similar to NF90 conserved binding motif in TPT1-AS1 sequence. (B) RIP assays were performed using NF90 antibody or IgG, specific primers were used to detect TPT1-AS1, GAPDH and VEGFA in HCT116 cells. (C) RNA pull-down assays were performed using TPT1-AS1 probe or antisense RNA. GAPDH was used as the control. (Di) and (Dii) RIP assays were performed using anti-NF90 or non-specific IgG in TPT1-AS1 or sh-TPT1-AS1 transfected CRC cells. The RIP-derived VEGFA mRNA was detected using qRT-PCR and presented as a percentage of the input. (E) The expression of NF90 and VEGFA were confirmed using western blot assays. (F) The half-life of VEGFA mRNA in indicated CRC cells were determined by qRT-PCR. (G) Transwell invasion assays were used to determine the invasive ability of indicated cells. (H) The CD31 expression of indicated xenograft tumors were detected by IHC staining. **P < 0.01, # represents no statistical significance.