Abstract

 
 
 
 Purpose: To determine the effect of miR-124 in pituitary prolactinoma.
 Methods: The viability and proliferation of prolactinoma cells were investigated using Cell Counting Kit-8 and 5-ethynyl-2′-deoxyuridine staining assays. Cell migration and invasion were investigated using the transwell assay. The epithelial-mesenchymal transition was investigated using western blotting. The target gene of miR-124 was verified by the luciferase activity assay.
 Results: The viability and proliferation of prolactinoma cells were repressed by miR-124 over-expression (p < 0.01). Forced miR-124 expression suppressed prolactinoma cell migration and invasion (p < 0.01). E-cadherin expression was enhanced, while N-cadherin and vimentin were reduced, by miR- 124 over-expression (p < 0.01). PHF19 (plant homeodomain-like finger protein 19) contains an miR-124 binding site, and PHF19 over-expression enhanced cell proliferation, promoted cell migration and invasion, reduced E-cadherin expression and enhanced N-cadherin and vimentin expression in prolactinoma cells. Additionally, miR-124 mimic-induced suppression of prolactinoma cell growth and metastasis was attenuated by forced PHF19 expression.
 Conclusion: MiR-124 retards prolactinoma cell growth and metastasis by reducing PHF19, providing a promising therapeutic target for prolactinoma.
 
 
 
Highlights
Pituitary adenomas are common intracranial neoplasms, accounting for 15 % of all intracranial tumors in China [1]
EdU-positive MMQ and GH3 cells transfected with miR-124 mimic were lower than those transfected with NC mimic (Figure 1 C), suggesting that forced miR-124 repressed pituitary prolactinoma cell proliferation
The suppressive effect of miR-124 on PHF19 expression and promotive effect of miR-124 silencing on PHF19 expression in MMQ and GH3 cells (Figure 3C) suggest that miR-124 directly binds to PHF19 in pituitary prolactinoma
Summary
Pituitary adenomas are common intracranial neoplasms, accounting for 15 % of all intracranial tumors in China [1]. The effect of miR-124 on prolactinoma has not yet been reported. We hypothesized that miR-124 might suppress prolactinoma cell growth and metastasis by down-regulating PHF19. The vectors were cotransfected with miR-124 mimic or inhibitor into HEK-293T cells, and the Dual-Luciferase Reporter Assay System (Promega, Madison, Wisconsin, USA) was used to determine the luciferase activity. MiR-124 mimic and inhibitor were synthesized by Invitrogen (Carlsbad, CA, USA). Cells (3 × 105 cells/well) were transfected with 20 nM of miR-124 mimic or inhibitor or 300 nM pcDNA-PHF19 (Invitrogen) using Lipofectamine 2000 (Invitrogen). Cells were seeded in 96-well plates for 24 h and incubated with medium containing 100 μL of 5ethynyl-2′-deoxyuridine EdU (50 μM; Invitrogen) for 12 h. The membranes were incubated with primary antibodies against PHF19, E-cadherin, N-.
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