PURPOSE: To investigate the effects of miR-1246 on proliferation, invasion, and migration in the human (CSCC) cell line SiHa. METHODS: SiHa cells were assigned into three groups: miR-1246 analog; miR-1246 antagonist; and control. The MTT, transwell, and wound healing assays were performed to evaluate the proliferation, invasion, and migration abilities of SiHa cells, respectively. Western blot was carried out to detect protein expression of thrombospondin-2 (THBS2) before and after transfection with miR-1246 analog, antagonist, or control. In addition, a THBS2 3'-UTR-containing dual luciferase plasmid was generated and co-transfected with miR-1246, the inhibitor, or non-specific miRNA, into SiHa cells to observe its effects on THBS2-driven luciferase enzyme activity. RESULTS: MTT, transwell, and wound healing assays revealed that proliferation, migration, and invasion were all significantly enhanced (P < 0.01) in SiHa cells transfected with miR-1246 analog, but were suppressed in those transfected with the miR-1246 antagonist. Western blot data showed that miR-1246 analog-transfected SiHa cells had significantly decreased THBS2 expression when compared with control-transfected cells (gray value = 6.28 ± 10.22 vs. 9.58 ± 17.58; P = 0.013) while those transfected with the miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90 ± 19.81; P = 0.037). Moreover, SiHa cells co-transfected with miR-1246 and the THBS2 3'-UTR-containing plasmid exhibited decreased luciferase enzyme activity compared with the control. CONCLUSION: MiR-1246 induced CSCC SiHa cell proliferation, invasion and migration. Preliminary evidence suggests that miR-1246 might promote CSCC tumorigenesis and progression by the suppression of its target gene THBS2.
PURPOSE: To investigate the effects of miR-1246 on proliferation, invasion, and migration in the human (CSCC) cell line SiHa. METHODS: SiHa cells were assigned into three groups: miR-1246 analog; miR-1246 antagonist; and control. The MTT, transwell, and wound healing assays were performed to evaluate the proliferation, invasion, and migration abilities of SiHa cells, respectively. Western blot was carried out to detect protein expression of thrombospondin-2 (THBS2) before and after transfection with miR-1246 analog, antagonist, or control. In addition, a THBS2 3'-UTR-containing dual luciferase plasmid was generated and co-transfected with miR-1246, the inhibitor, or non-specific miRNA, into SiHa cells to observe its effects on THBS2-driven luciferase enzyme activity. RESULTS:MTT, transwell, and wound healing assays revealed that proliferation, migration, and invasion were all significantly enhanced (P < 0.01) in SiHa cells transfected with miR-1246 analog, but were suppressed in those transfected with the miR-1246 antagonist. Western blot data showed that miR-1246 analog-transfected SiHa cells had significantly decreased THBS2 expression when compared with control-transfected cells (gray value = 6.28 ± 10.22 vs. 9.58 ± 17.58; P = 0.013) while those transfected with the miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90 ± 19.81; P = 0.037). Moreover, SiHa cells co-transfected with miR-1246 and the THBS2 3'-UTR-containing plasmid exhibited decreased luciferase enzyme activity compared with the control. CONCLUSION:MiR-1246 induced CSCC SiHa cell proliferation, invasion and migration. Preliminary evidence suggests that miR-1246 might promote CSCC tumorigenesis and progression by the suppression of its target gene THBS2.
Authors: S Hasegawa; H Eguchi; H Nagano; M Konno; Y Tomimaru; H Wada; N Hama; K Kawamoto; S Kobayashi; N Nishida; J Koseki; T Nishimura; N Gotoh; S Ohno; N Yabuta; H Nojima; M Mori; Y Doki; H Ishii Journal: Br J Cancer Date: 2014-08-12 Impact factor: 7.640