PMC

DAB2 was identified as a miR-106b-directed target gene. a The predicted interaction site of miR-106b and candidate target gene DAB2 3′UTR. b Luciferase assay of HEK-293A cells co-transfected with miR-106b mimic and pGL-3-DAB2 plasmid (miR-NC and miR-106b with DAB2-wt 3′UTR; miR-NC and miR-106b with DAB2-mut 3′UTR) after 24 h. Data are mean ± SEM(n = 3).**P < 0.01

Knockdown of miR-106b inhibited TGF-β1-induced cell migration in cervical carcinoma. a HeLa cells were transfected with 100-nm miR-106b inhibitor (anti-miR-106b) or miR-106b control inhibitor (anti-NC) for 24 h, then migration was induced with or without TGF-β1 (5 ng/mL) for 24 h. b Quantification of A.**P < 0.01, ^$$ P < 0.01, ^# P < 0.05. c SiHa cells were transfected with 100-nm miR-106b inhibitor (anti-miR-106b) or miR-106b control inhibitor (anti-NC) for 24 h, then migration was induced with or without TGF-β1 (5 ng/mL) for 24 h. d Quantification of C.**P < 0.01, ^$$ P < 0.01, ^# P < 0.05

miR-106b is involved in TGF-β1-induced cell migration by targeting DAB2 in cervical carcinoma. The Large circle represents one cervical cancer cell. The orange small circles represent many TGF-β1 cell factors. The two small rectangular blocks on the circle indicate the receptors of TGF-β1. The red rectangle represents miR-106b. The blue oval represents DAB2 protein

miR-106b promoted the migration of SiHa cells tested by scratch and transwell assay. SiHa cells were transfected with a miR-106b inhibitor (anti-miR-106b) at 100 nm. and b miR-106b mimics (miR-106b) at 50 nm. Scratch assay of cell migration with c miR-106b knockdown and d miR-106b overexpression. Transwell migration assay with e miR-106b knockdown and f miR-106b overexpression. Data are mean ± SEM (n = 3).*P < 0.05, **P < 0.01 compared to control

Knockdown of DAB2 promoted TGF-β1-induced cell migration in cervical carcinoma. a Scratch assay of cell migration in cells transfected with DAB2-NC or DAB2 siRNA at 0 and 24 h. Data are mean ± SEM (n = 3).*P < 0.05 compared to control. b Transwell migration assay of cells transfected with DAB2-NC or DAB2 siRNA at 24 h. Data are mean ± SEM(n = 3).^* P < 0.01 compared to control. c HeLa cells were transfected with DAB2-NC or DAB2 siRNA for 24 h, then migration was induced with or without TGF-β1 (5 ng/mL) for 24 h and quantification. d Quantification of C. **P < 0.01, ^$ P < 0.05, ^# P < 0.05

miR-106b promoted the migration of HeLa cells tested by scratch and transwell assay. HeLa cells were transfected with a miR-106b inhibitor (anti-miR-106b) at 100 nm and b miR-106b mimic(miR-106b) at 50 nm. Scratch assay of cell migration with c miR-106b knockdown and d miR-106b overexpression. Scratched cells were observed and photographed at 0 and 24 h under a microscope. Data are mean ± SEM(n = 3).*P < 0.05,**P < 0.01 compared to control. Transwell migration assay with e miR-106b knockdown and f miR-106b overexpression. Cells were transfected with miR-106b inhibitor(anti-miR-106b) and miR-106b control inhibitor (anti-NC). The cells on the transwell chambers were fixed, dyed, observed and photographed after culture for 24 h. Data are mean ± SEM(n = 3). *P < 0.05 compared to control

DAB2, a predicted target gene of miR-106b, was inhibited by TGF-β1 in part via miR-106b. a Real-time PCR analysis of mRNA expression of miR-106b in HeLa cells with TGF-β1 treatment for 24 h. Data are mean ± SEM (n = 3). ^* P < 0.01 compared to control. b Western blot analysis of protein level of DAB2 in HeLa cells transfected with miR-106b inhibitor (anti-miR-106b) or miR-106b control inhibitor (anti-NC). Data are mean ± SEM (n = 3). *P < 0.05 compared to control. c Western blot analysis of DAB2 protein level in HeLa cells transfected with 100 nm miR-106b inhibitor (anti-miR-106b) for 24 h, then with or without TGF-β1 (5 ng/mL) for 24 h; GAPDH was an internal control. d Quantification of C.*P < 0.05

High expression of miR-106b in human cervical tissues detected by Q-PCR and in situ hybridization. a Real-time PCR analysis of the mRNA expression of miR-106b in 19 human cervical carcinoma tissues and 19 normal cervical samples. U6 small nuclear RNA was an internal control. Each point represents 1 sample. The horizontal bar is the mean and whiskers are SEM. ^** P < 0.01 (Mann–Whitney test). b In situ hybridization of miR-106b in negative control and normal cervical specimens and cervical carcinoma tissue. c Quantification of  In situ hybridization staining of 19 normal cervical specimens and 19 cervical carcinoma tissues

Overexpression or knockdown of miR-106b with mimic or inhibitor in cervical cancer cells. a Real-time PCR of the expression of miR-106b in 4 human cervical cancer cell lines. The expression of miR-106b in C33A cells was a control. *P < 0.05, **P < 0.01, *** P < 0.001 compared with vehicle. Data are mean ± SEM (n = 3). b miR-106b mimic (miR-106b) at four concentrations (10, 50, 100, 200 nm) was transfected into HeLa cells to overexpress miR-106b. miR-NC was a negative control.**P < 0.01, ***P < 0.001 compared with vehicle. Data are mean ± SEM (n = 3). c miR-106b inhibitor (anti-miR-106b) at four concentrations (50, 100, 150, 200 nm) was transfected into HeLa cells to knockdown miR-106b. anti- NC was a negative control.**P < 0.01, ***P < 0.001compared with vehicle. Data are mean ± SEM (n = 3)

DAB2 had low expression in cervical cancer tissues, and which was negatively correlated with miR-106b expression. a The expression of DAB2 in 19 human cervical carcinoma tissues and 19 normal cervical tissues (**P < 0.01) (nonparametric test with Mann–Whitney test). U6 small nuclear RNA was an internal control. Each point represents 1 sample. The horizontal bar is the mean and whiskers are SEM. b Spearman correlation analysis of DAB2 and miR-106b expression in 19 cervical cancer tissues.***P < 0.0001, R2 = −0.7744. c Immunohistochemistry of protein expression of DAB2 in cervical tissues. Expression of DAB2 protein in a) normal ovarian tissue as a positive control, b) normal cervical tissues, c) cervical carcinoma tissues, and d) negative control. d Quantification of immunohistochemical staining of 19 normal cervical specimens and 19 cervical carcinoma tissues.**P < 0.01