Figures & data
Figure 1. lncRNA TTTY15 sponges miR-98-5p in gastric cancer.
(A) starBase predicted the binding sites between miR-98-5p and lncRNA TTTY5. (B) Dual-luciferase reporter assay verified the target relationship between miR-98-5p and lncRNA TTTY5. (C) RNA pull-down analysis revealed the targeted relationship between miR-98-5p and lncRNA TTTY5.
Figure 2. Ectopic expressions of lncRNA TTTY5 and miR-98-5p in gastric cancer tissues and cells.
(A) lncRNA TTTY5 levels in tumor tissues and non-tumor tissues obtained from 30 patients with gastric cancer. (B) Aberrant expressions of lncRNA TTTY5 in the GES-1 cell line and gastric cancer cell lines (AGS, SNU-5, and NCI-N87). (C) miR-98-5p levels in tumor tissues and non-tumor tissues obtained from 30 patients with gastric cancer. (D) Aberrant expressions of miR-98-5p in the GES-1 cell line and gastric cancer cell lines (AGS, SNU-5, and NCI-N87).
Figure 3. lncRNA TTTY5 suppressed miR-98-5p expression in AGS cells to regulate AGS cell proliferation.
(A) si-TTTY15 greatly knocked down lncRNA TTTY5 expression in AGS cells. (B) Transfection efficacy of miR-98-5p after miR-98-5p inhibitor interference treatment in AGS cells. (C) miR-98-5p level in AGS cells after transfection with si-TTTY15 and/or miR-98-5p inhibitor. (D) lncRNA TTTY5 silencing greatly reduced AGS cell proliferation. lncRNA TTTY5 silencing decreased the expression of PCNA protein (E) and the expression of PCNA mRNA (F) in AGS cells.
Figure 4. lncRNA TTTY5 induced AGS cell apoptosis by sponging miR-98-5p.
(A) Flow cytometry analysis of the effect of lncRNA TTTY5 and miR-98-5p on AGS cell apoptosis (B) Quantified results of the flow cytometry analysis. (C) lncRNA TTTY5 silencing promoted caspase-3 activity in AGS cells by sponging miR-98-5p. Cleaved-caspase-3 protein (D) and the cleaved caspase-3/GAPDH ratio (E) in AGS cells transiently transfected with si-TTTY-5 and miR-98-5p inhibitor.
Figure 5. Identification of CCND2 as a downstream target of miR-98-5p in gastric cancer.
(A) The TargetScan bioinformatics tool predicted the binding sequences between miR-98-5p and CCND2. (B) Dual-luciferase reporter assay confirmed the target relationship between miR-98-5p and CCND2. (C) qRT-PCR analysis was used to determine CCND2 expression in tumor and non-tumor tissues obtained from 30 gastric cancer patients. (D) Ectopic expressions of CCND2 in the GES-1 cell line and gastric cancer cell lines.
Figure 6. miR-98-5p negatively regulated CCND2 expression in AGS cells.
(A) Transfection efficacy of miR-98-5p in AGS cells. (B) Overexpression of the CCND2-plasmid construct in AGS cells. The impact of miR-98-5p transfection on the expression of CCND2 protein (C) and mRNA (D) in AGS cells.
Figure 7. miR-98-5p overexpression suppressed AGS cell proliferation and promoted apoptosis by targeting CCND2.
(A) AGS cell proliferation was assayed by MTT assay. (B) Western blotting was used to ascertain the expression of PCNA protein. (C) The PCNA mRNA level was confirmed by PCR analysis. (D-E) Flow cytometry analysis of the AGS cell apoptosis rate. (F) Caspase-3 activity in different groups. The level of cleaved caspase-3 protein (G) and the cleaved-caspase-3/GAPDH ratio (H) were evaluated by Western blot assay.
