Fig 2: Targeting relationship between miR‐1254 and CSF‐1 in glioma. (A) The predicted base pairing in miR‐1254 and CSF‐1 from TargetScan. (B) Validation of targeting relation between miR‐1254 and CSF‐1 through dual‐luciferase reporter assay. (C) The expression of CSF‐1 was higher in glioma cell lines than that in normal cell line NHAs. (D) CSF‐1 protein expression was higher in glioma cell lines than that in normal cell line NHAs. (E) Relative CSF‐1 expression in glioma samples divided according to pathological classification and NBTs. The Student's t test was used to analyse significant differences between groups. (F) TCGA database showing increased CSF‐1 expression in glioma compared with normal samples. (G) Western blotting of CSF‐1 in U87 and U251 cells transfected with indicated miR‐1254 or anti‐miR‐1254. Results were expressed as means ± SD of three independent experiments. *P < .05; **P < .01

Fig 3: CSF‐1 promoted the proliferation, colony formation, migration and invasion of glioma. (A) U87 cells of CSF‐1 overexpression and knockdown were analysed by real‐time PCR. (B) CCK‐8 assay showed that cell viability was significantly stronger in CSF‐1 overexpression group while was weaker in CSF‐1 knockdown group, and there was no significant difference between CSF‐1‐KD + anti‐miR‐1254 group and mock group. (C) and (D) Colony formation assay showed that colony number in CSF‐1 overexpression group was remarkably higher, whereas that in CSF‐1 knockdown group was much lower, and there was no significant difference between CSF‐1‐KD + anti‐miR‐1254 group and mock group. (E) and (F) Wound‐healing assay showed that cell migration rate in CSF‐1 overexpression group was remarkably higher, whereas that in CSF‐1 knockdown group was much lower, and there was no significant difference between CSF‐1‐KD + anti‐miR‐1254 group and mock group. (G) and (H) More invaded cells were observed in CSF‐1 overexpression group while the number of invaded cells in CSF‐1 knockdown group was significantly lower compared with the mock group, and there was no significant difference between CSF‐1‐KD + anti‐miR‐1254 group and mock group. (I) Cell apoptosis rate was notably higher in CSF‐1 knockdown group while lower in CSF‐1 overexpression group, and there was no significant difference between CSF‐1‐KD + anti‐miR‐1254 group and mock group. Results were expressed as means ± SD of three independent experiments. *P < .05; **P < .01