Figures & data
Figure 1. High expression of LINC01133 accelerated the malignant phenotypes of CC cells. (a) Differential expression of LINC01133 in CC was obtained from The Cancer Genome Atlas (TCGA). (b) Relative to normal cervical cells, LINC01133 was markedly high-expressed in CC cells by qRT-PCR. (c) The expression of LINC01133 in cells transfected with shLINC01133#1, shLINC01133#2 and shLINC01133#3 were examined. (d-e) LINC01133 inhibition suppressed the growth of ME-180 and MS751 by CCK-8 and EdU assay. (f) LINC01133 silencing posed pro-apoptosis effect detected by caspase-3 activity assay. *P < .05, **P < .01.
Figure 2. LINC01133 elevated the invasive and migratory capacity of CC cells and accelerated EMT. (a) LINC01133 down-regulation impeded migratory potential of ME-180 and MS751 by Transwell migration assay. (b) Transwell invasion assay indicated the invasion-inhibiting effect of LINC01133 suppression. (c) LINC01133 interference hindered EMT process in CC. *P < .05.
Figure 3. LINC01133 directly targeted miR-4784. (a) The location of LINC01133 was discovered in ME-180 and MS751 cells. (b) The expression of 15 potential miRNAs predicted by bioinformatics tools was examined in ME-180 cells by qRT-PCR after LINC01133 blockade. (c) Potential binding site of LINC01133 for miR-4784 was predicted. (d) Luciferase reporter assay indicated a suppressed activity of wild type of LINC01133 3′-UTR reporter in ME-180 and MS751 cells in the presence of miR-4784 mimics. (e) Cell lysate of ME-180 and MS751 cell lysates underwent incubation with anti-IgG or anti-Ago2 antibody and the enriched LINC01133 and miR-4784 were analyzed by qRT-PCR. *P < .05, **P < .01, *** P < .001.
Figure 4. LINC01133 up-regulated the target gene of miR-4784, AHDC1. (a) Bioinformatic analysis of five potential downstream genes of miR-4784. (b) The expression of AHDC1 was detected in normal cervical cells and CC cells. (c) The predicated binding sites between AHDC1 and miR-4784. (d) The expression level of LINC01133 post-transfection of pcDNA3.1/LINC01133 into ME-180 and MS751 cells was tested. (e) The luciferase activity of AHDC1-WT reporter was monitored after transfection with specific plasmids. (f) RIP assay consolidated the binding between AHDC1 and miR-4784. (g-h) qRT-PCR and western blot disclosed the effect of LINC01133 or miR-4784 on AHDC1 mRNA and protein levels. *P < .05, **P < .01, *** P < .001.
Figure 5. MiR-4784/AHDC1 reversed the modulatory impact of LINC01133 on invasive and migratory potentials and EMT in CC. (a) Confirmation of the overexpression efficiency of AHDC1 in ME-180 and MS751 cells was accomplished by qRT-PCR. (b-c) Post-transfection of indicated plasmids, the growth of ME-180 cells was assessed by CCK-8 and EdU assays. (d) ME-180 cell apoptosis after transfection with certain plasmids was evaluated by caspase-3 activity assay. (e-f) Transwell assay was executed to probe the migration and invasion of ME-180 cells transfected with specific plasmids. (g) Western blot was implemented to examine the effects of LINC01133/miR-4784/AHDC1 pathway on the EMT-associated protein contents. *P < .05, **P < .01.
