Skip to main content
. 2022 Aug 4;2022:4156966. doi: 10.1155/2022/4156966

Figure 6.

Figure 6

hsa-miR-1254 binds KRT80 and HNSCAT1. (a) Through TargetScan and miRDB, we found 268 miRNAs that may serve as binding candidates for HNSCAT1. Notably, 4 of them (miR-1254, miR-3150b-3p, miR-4505, and miR-149-3p) were predicted to interact with the 3′UTR of KRT80 mRNA. (b, c) Real-time PCR was performed to observe KRT80 expression after transfecting miRNA mimics (miR-1254, miR-3150b-3p, miR-4505, and miR-149-3p) into HNSCAT1-overexpressing cells. Experiments were conducted in triplicate, and the results are shown as the mean ± SEM. ∗p < 0.05. (d) Western blotting assays were conducted to observe KRT80 protein levels after transfecting miRNA mimics into HNSCAT1-overexpressing cells. (e) Schematic figure of the miRNA-binding RNA capture assay. (f, g) Real-time PCR was performed on these miRNA-binding RNAs. The miRNA probe transfection-free group (beads group) was set to 1. These results indicated that miR-1254 interacts with (f) KRT80 and (g) lincRNA HNSCAT1. (h) A reporter gene assay demonstrated that miR-1254 mimics significantly inhibited reporter activity, while the miRNA potential binding site-mutated group (lanes 4-6) and miR-1254 inhibitor-treated group (lane 3) presented similar reporter signals compared to the control group. Experiments were conducted in triplicate, and the results are shown as the mean ± SEM. ∗p < 0.05.