Over-expression of miR-204 suppresses proliferation and invasion of retinoblastoma cells and its possible mechanism
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Abstract:
Objective: To observe the effects of miR-204 on the proliferation and invasion of retinoblastoma (RB) cells and to explore the potential regulatory mechanism. Methods: The expression level of miR-204 in RB cell lines (Y79, SO-RB50, and HXO-Rb44) as well as in normal human retinal pigment epithelial cell line hTERT RPE-1 was detected using qRT-PCR. The Y79 cells were divided into two groups (negative control group and miR-204 group) by respectively transfecting Y79 cells with NC-mimics and miR-204 mimics using liposome transfection method. The effects of miR-204 on Y79 cell proliferation was detected with CCK-8 assay; while the effect of miR-204 on migration and invasion of Y79 cells were determined by cell scratch assay and Transwell assay, respectively. Besides,the potential target gene of miR-204 was predicted by bioinformatics; and the influence of miR-204 on the expression of high mobility group AT-hook 2 gene (HMGA2) at both mRNA and protein levels was detected using qRT-PCR and Western blotting, respectively.Results: miR-204 expression in RB cell lines Y79, SO-RB50 and HXO-Rb44 was remarkably lower than that in normal human retinal pigment epithelial cell line hTERT RPE-1 (P<0.01). miR-204 expression in Y79 cells was markedly up-regulated after transfection with miR-204 mimics (P<0.01) along with significantly reduced cell proliferation, migration and invasion capacities (all P<0.01), and mRNA and protein expressions of HMGA2 were also outstandingly reduced (P<0.01). Conclusion: miR-204 is lowly expressed in RB cell lines; in addition, miR-204 over-expression can suppress RB cell proliferation, migration and invasion, the mechanism of which might be related to down-regulation of the expression of HMGA2.
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Project supported by the Medical and Health Science and Technology Depvolopment Project of Shangdong Province (No.2017WS751)