Fig 1: Compound 5 inhibits metastasis of TNBC in vivo. (A) In vivo treatment of 5 (10 mg/kg, q.o.d., 6 wk) decreased lung nodules (white nodules) stained with Bouin’s solution. (B) H&E staining of lung tissue from mice treated with 5 or vehicle. (C) Lung nodule tissue treated with 5, but not vehicle, exhibited decreased mature miR-21 and (D) pre–miR-21 expression, as measured by FISH probing. (E) Treatment with 5 increased PDCD4 levels in lung nodule tissue sections as determined by immunohistochemistry (IHC) staining. (Scale bar, 50 µm.) Data represent mean ± SEM (n ≥ 4). *P < 0.05; **P < 0.01 as determined by a two-tailed Student t test.
Fig 2: Graphic abstract: circ_0008259 functions as a ceRNA to regulate the expression of miR-21-5p and PDCD4 to block OS progression.
Fig 3: Circ_0008259 represses OS progression via modulating miR-21-5p / PDCD4 axis. (A) 143B cells with circ_0008259 overexpression was transfected with miR-21-5p mimics and PDCD4 overexpression vector, and the expression of PDCD4 in OS cells was detected by Western blot assay. (B) CCK8 assay was used to detect the proliferation of 143B cells after transfection. (C, D) Transwell assay was used to detect the migration and invasion of 143B cells after transfection. (E) Flow cytometry was used to detect the apoptosis rate of 143B cells after transfection. **P < 0.01, and ***P < 0.001.
Fig 4: The expression profiles of seven target genes that may be regulated by miRNA-96-5p obtained from the GEPIA website in rectal cancer tissues and normal tissues. The median values of CAV1, DAB2, DDIT3, FOXO3, GPC3, MBD4 and PDCD4 in rectal cancer tissues and normal tissues were 17.16 and 180.87, 23.78 and 22.54, 27.78 and 33.00, 19.58 and 15.76, 1.24 and 4.3, 17.76 and 18.13, 92.51 vs.110.35, respectively. The red line represents the tumor, and the green line represents normal tissue.
Fig 5: Expression of predicted target genes of miRNAs in lung tissues of asbestos‐exposed mice and correlation with miRNA expression. (A) Expression of predicted target genes of miR‐21 and miR‐133a. Expression levels of predicted target genes of miR‐21 (Pdcd4 and Reck) and miR‐133a (Gdnf) were quantified by real‐time PCR as described in Section 2. Gapdh was used as an internal control. Data were expressed as means ± SD of 4–5 independent experiments. *P < .05, **P < .01 and ***P < .001 compared with control; # P < .05 and ## P < .01, between chrysotile and crocidolite by ANOVA followed by Tukey’s test. (B) Correlation of the expression levels of miRNA and predicted target genes. Individual points show the expression levels of mRNA and corresponding target gene in each lung tissue. The correlations of the expression levels of miRNAs and target genes were analyzed by Pearson’s correlation test. ANOVA, analysis of variance; miRNA, microRNA; PCR, polymerase chain reaction; PDCD4, programmed cell death 4; RECK, reversion‐inducing‐cysteine‐rich protein with kazal motifs; SD, standard deviation
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