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. 2021 Feb 16;10:570916. doi: 10.3389/fonc.2020.570916

Figure 4.

Figure 4

tRF-3017A may play a role in regulating the migration and invasion of GC cells by regulating NELL2. (A) Venn diagram evaluated the overlapped genes among miRanda, TargetScan, TargetRank, and tRFTar predictions. (B, C) The expression levels of eight predicted target genes were performed to detect in AGS cell line after transfection with tRF-3017A-inhibitor or mimics by qRT-PCR. (D, E) The expression levels of NELL2 were detected in AGS cell line after transfection with tRF-3017A-inhibitor or mimics by western blot. (F) GC cell migration and invasion ability were detected after transfected with si-NELL2 and pEX3-NELL2. (G) Expression of NELL2 was downregulated (-ΔCt, paired t test) in GC tissues relative to matched-paired NATs among 84 GC patients by qRT-PCR. (H) Correlation analysis of relative expressions of tRF-3017A and NELL2 and “r” is the correlation coefficient. (I) The luciferase activity of wild type NELL2 3’UTR or mutant NELL2 3’UTR after transfection with tRF-3017A mimics or 3017A-inhibitor and their corresponding NC in AGS cell line. (J) The result of the RIP based on Ago2 showed that tRF-3017A may exert its miRNA-like silencing effect by combining Ago2, thus targeting NELL2. Representative images and bar graphs were depicted. Data are shown as mean ± SD. NC, negative control. *P < 0.05; **P < 0.01.