Fig. 3

miR-138-5p is involved in RvD1-mediated suppression of cell growth and invasion. a. Heat map diagram with supervised hierarchical clustering of miRNAs expressed by RvD1-treated A549 cells. The miRNA clustering tree is shown on the left, and the sample clustering tree appears at the top (control and RvD1 200 μg/L group). The color scale in the map illustrates the relative expression level of a miRNA across all samples: red represents an expression level above the mean, and blue represents expression lower than the mean. b. miR-138 miRNA expression data were downloaded from the TCGA database containing 344 LUSC and 327 LUAC cancer tissues. The overall survival (OS) of patients with tumors expressing high vs. low miR-138 was analyzed by the Cox regression test. The relationship between miR-138-5p and tumor extent (T1-T4) or distant metastasis (M0-M1) was analyzed by Kruskal-Wallis test. c. A549 cells were transfected with control mimics (Con), miR-138-5p mimics, control anti-sense RNA inhibitor (Negcon), or miR-138 inhibitor, and then subjected to cell growth and invasion assays. *P < 0.05, **P < 0.01 compared to Con group; ^##P < 0.01 compared to Negcon group. d. A549 cells were transfected with Negcon or miR-138 inhibitor, and subjected to cell growth and invasion assays (n = 4). **P < 0.01 compared to Negcon group; ^#P < 0.05, ^##P < 0.01 compared to NegCon+RvD1 group. N.S. no significance. There is no significant difference in miR-138 inhibitor-treated groups with or without RvD1 treatment (P = 0.052). e. A549 cells were treated as above and subjected to invasion assays. There is no significant difference in miR-138 inhibitor-treated groups with or without RvD1 treatment (P = 0.077). The transwell units were stained and high power views (200×) were selected randomly from each sample in a blind manner. f. A549 cells were treated as above. Akt and Erk1/2 phosphorylation were evaluated by Western blot analysis using total Akt and Erk1/2 as internal controls. *P < 0.05, **P < 0.01 for p-Akt compared to Con group; ^#P < 0.05, ^##P < 0.01 for p-Erk1/2 compared to Con group