Figure 2.

EVs-miR-181b-5p Is Highly Expressed in ESCC and Mediates Angiogenesis In Vitro

(A) Microarray analysis of significantly expressed miRNAs in ESCC tissues and adjacent normal tissue miRNA from seven ESCC patients was presented in a heatmap. (B) Hierarchical clustering analysis of 10 of the most upregulated and 21 of the most downregulated miRNAs (fold change > 2.5 or < −2.5; p < 0.001) was shown. (C) The expression of miR-181b-5p in 58 pairs of ESCC tissues was detected by qRT-PCR. ΔΔCt=–((Ct_miR-181b-5p-Ct_U6)_ESCC-(Ct_miR-181b-5p-Ct_U6)corresponding non-malignant tissue). (D) The expression of miR-181b-5p in 10 ESCC cell lines compared with normal esophageal epithelia cells (HEEC) was detected by quantitative relative real-time PCR. (E) The expression of miR-181b-5p in EVs from the cell lines referred above was detected by quantitative relative real-time PCR. (F) Significant correlation of miR-181b-5p expression in ESCC cells and ESCC cell-derived EVs was calculated with Spearman’s test. (G–O) Migration (G and K), wound healing (H and L), tube formation (I and M), cell cycle (J and N), and cell viability (O) assays of HUVECs treated with miR-181b-5p mimic, inhibitor, or negative control in vitro. Representative images and quantitative analysis were shown. Scale bars, 200 μm. Each experiment was performed three times independently, and data are shown as mean ± SD. Student’s t test was used to analyze the data (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001).