目的:观察miR-204 对成视网膜细胞瘤（retinoblastoma，RB）细胞增殖与侵袭的影响，探讨其可能的调控机制。方法:应用实时荧光定量PCR（qRT-PCR）检测RB细胞系Y79、SO-RB50、HXO-Rb44 和人正常视网膜色素上皮细胞系hTERT RPE-1 中miR-204 的表达水平。将Y79 细胞系分成阴性对照组和miR-204 组，分别应用脂质体转染法转染NC-mimics 和miR-204 mimics，CCK-8 增殖实验检测miR-204 表达对Y79 细胞增殖的影响，细胞划痕实验和Transwell 小室法检测miR-204 对Y79 细胞迁移和侵袭的影响，应用生物学信息法预测miR-204 的可能作用靶基因，应用qRT-PCR 和Western blotting 检测miR-204 对靶基因高迁移率族蛋白A2（high mobility group AT-hook 2，HMGA2）mRNA和蛋白表达的影响。结果: miR-204 在RB 细胞系Y79、SO-RB50、HXO-Rb44 中的表达较人正常视网膜色素上皮细胞系hTERT RPE-1 明显降低（P<0.01）。转染miR-204 mimics 后，Y79 细胞中miR-204 表达明显升高（P<0.01）、细胞增殖能力明显下降（P<0.01）、迁移及侵袭能力明显降低（P<0.01），miR-204 的靶基因HMGA2 mRNA和蛋白的表达明显下降（P<0.01）。结论: miR-204 在RB细胞系中低表达，过表达miR-204 能够抑制RB细胞的增殖、迁移及侵袭能力，其机制可能与下调HMGA2 基因的表达有关。

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.

山东省医药卫生科技发展项目（No. 2017WS751)