The expression of miR-503 was down-regulated in osteosarcoma (A)qRT-PCR analysis of miR-503 expression in 30 pair'sosteosarcoma tissues and their corresponding adjacent normal tissues.

<p>The expression of miR-503 was normalized to U6 snRNA. (B) Relative miR-503expressionlevels inosteosarcoma tissues and their corresponding adjacent normal tissues. (C) Loss of miR-503 levels in patients with osteosarcoma was associated with associated with considerably shortened disease-free survival. (D) The relative expression levels were determined by qRT-PCR in Human osteosarcoma cell lines (MG-63, U2OS, SOSP-9607, and SAOS-2)and hFOB. ***p<0.001.</p

Overexpression of miR-503inhibitedmigration and invasion of osteosarcoma cells.

<p>(A) Migration assays of the MG-63cells after treatment with miRNA mimics, inhibitors or scramble or no transfection; the relative ratio of migratory cells per field is shown on the right. (B) Invasion analysis of the MG-63 cells after treatment withmiRNA mimics, inhibitors or scramble or no transfection; the relative ratio of invasive cells per field is shownon the right. (C) Migration assays of the U2OScells after treatment with miRNA mimics, inhibitors or scramble or no transfection; the relative ratio of migratory cells per field is shown on the right. (D) Invasion analysis of the U2OScells after treatment withmiRNA mimics, inhibitors or scramble or no transfection; the relative ratio of invasive cells per field is shownon the right, *p<0.05, ** p<0.01, and ***p<0.001.</p

L1CAM was inversely expressed with miR-503 in osteosarcoma (A) qRT-PCR analysis of L1CAM expression in 30 pair's osteosarcoma tissues and their corresponding adjacent normal tissues.

<p>The expression of L1CAM was normalized to GAPDH. (B) Relative L1CAMexpressionlevels inosteosarcoma tissues and their corresponding adjacent normal tissues.(C) The L1CAM relative mRNA expression levels were determined by qRT-PCR in Human osteosarcoma cell lines (MG-63, U2OS, SOSP-9607, and SAOS-2)) and hFOB. (D) Western blot analysis ofL1CAM expression in Human osteosarcoma cell lines (MG-63, U2OS, SOSP-9607, and SAOS-2)and hFOB. (D) Analysis of correlation of miR-503 and L1CAM expression in osteosarcoma tissues. (Two-tailed Pearson's correlation analysis, r = −0.86; p<0.01, n = 30). Data was presented as log 2 of fold change of osteosarcoma tissues relative to non-tumor adjacent tissues.***p<0.001.</p

miR-503 targeted L1CAM in osteosarcoma cells.

<p>(A) The sequences of miR-503 binding sites within the human L1CAM 3'UTRs and schematic reporter constructs, in this panel, L1CAM -WT represent the reporter constructs containing the entire 3'UTR sequences of L1CAM. L1CAM -MUT represent the reporter constructs containing mutated nucleotides. (B) The analysis of the relative luciferase activities of L1CAM -WT, L1CAM -MUT. The error bars are derived from triplicate expriments. (C) qRT-PCR analysis of L1CAM mRNA expression in the Mg-63 cells after treatment withmiRNA mimics or scramble or no transfection. The expression of L1CAM was normalized to GAPDH. (D) Western blot analysis ofL1CAM expression in the MG-63 cells transfected with miR-503mimics or scramble or no transfection. GAPDH was also detected as a loading control.***p<0.001.</p

Additional file 1: of Galectin-1 from cancer-associated fibroblasts induces epithelial–mesenchymal transition through β1 integrin-mediated upregulation of Gli1 in gastric cancer

Supplementary Table. Association of Gal-1 and Gli1 with clinicopathological indicators: age and gender. (DOC 33 kb