Research Paper Volume 11, Issue 24 pp 12114—12130

Reduced miR-203 predicts metastasis and poor survival in esophageal carcinoma

Figure 6. miR-203 promotes nuclear expression of β-catenin via enhancing Axin2 expression. (A) Proteins interacted with AXIN2 and KIF5C was predicted by String database (http://string-db.org). (B) AXIN2 mRNA expression level in tumor tissues and adjacent normal tissues of EC patients. Data is presented as mean ± SD. ***p < 0.001 (vs. adjacent normal tissues). (C) Scatterplot depicts a significant inverse and positive correlation between AXIN2 and KIF5C, miR-203 mRNA expression, respectively. (D) KYSE510 cells were transfected with miR-203 mimic or KIF5C recombinant plasmid. 48 h later, protein expression of Axin2, β-catenin in different cellular components were detected by western blotting. (E) AXIN2 mRNA expression in response to miR-203 mimic and KIF5C overexpression was determined by qRT-PCR. (F) Transcriptional activity of β-catenin has been determined by luciferase reporter gene assay. Data is presented as mean ± SD. ***p < 0.001 (miR-NC/KIF5C vs. miR-NC/Empty vector), ###p < 0.001 (miR-203/KIF5C vs. miR-203/Empty vector). (G) In some cases, KYSE510 cells were pretreated with IWR-1-endo (β-catenin pathway inhibitor) for 1 h, and then transfected with miR-203 mimic or KIF5C-expressing plasmid for 48 h. mRNA expressions of E-caherin, N-cadherin, MMP2 and MMP9 were detected by qRT-PCR. Datas are displayed as the Mean ± SD of three independent experiments.