Research Paper Volume 13, Issue 16 pp 20302—20318

Regulation of follistatin-like 3 expression by miR-486-5p modulates gastric cancer cell proliferation, migration and tumor progression

FSTL3 mRNA is regulated by miR-486-5p levels. (A) Schematic depiction of the predicted binding site for miR-486-5p in the FSTL3 mRNA. (B) Analysis of miR-486-5p levels in gastric cancer tumors vs. normal tissues in the STAD database in TCGA. (C) Correlative analysis of miR-486-5p and FSTL3 levels in gastric cancer tumors using the STAD database. (D) Profiling of miR-486-5p levels in normal and cancer gastric cancer cell lines using qRT-PCR. (E) Western blotting was used to analyze protein and RNA levels in gastric cancer MGC-803 cells subjected to transfection with miR-486-5p mimic or miR-486-5p inhibitor. (F) A dual luciferase assay was carried out using transfection of luciferase plasmid constructs alongside miR-486-5p mimic or miR-486-5p inhibitor. HEK 293T cells co-transfected with Luc-FSTL3-wt (WT) or Luc-FSTL3-mut (MUT) plasmids with synthetic miR-486-5p constructs. Error bars indicate +SEM; significance indicated by asterisks, *PPP

Figure 5. FSTL3 mRNA is regulated by miR-486-5p levels. (A) Schematic depiction of the predicted binding site for miR-486-5p in the FSTL3 mRNA. (B) Analysis of miR-486-5p levels in gastric cancer tumors vs. normal tissues in the STAD database in TCGA. (C) Correlative analysis of miR-486-5p and FSTL3 levels in gastric cancer tumors using the STAD database. (D) Profiling of miR-486-5p levels in normal and cancer gastric cancer cell lines using qRT-PCR. (E) Western blotting was used to analyze protein and RNA levels in gastric cancer MGC-803 cells subjected to transfection with miR-486-5p mimic or miR-486-5p inhibitor. (F) A dual luciferase assay was carried out using transfection of luciferase plasmid constructs alongside miR-486-5p mimic or miR-486-5p inhibitor. HEK 293T cells co-transfected with Luc-FSTL3-wt (WT) or Luc-FSTL3-mut (MUT) plasmids with synthetic miR-486-5p constructs. Error bars indicate +SEM; significance indicated by asterisks, *P<0.05, **P<0.01, ***P<0.001; number of experiments, n=3.