【摘要】 目的 探讨长链非编码RNA FGD5-AS1对垂体瘤（PA）细胞增殖、迁移、侵袭的影响，并分析其可能的作用机制。方法 体外培养人PA细胞系HPAs、RC-4BC、HP75和人星形胶质细胞系NHA,RT-PCR检测上述细胞系中FGD5-AS1和miR-15a的表达水平。取对数生长期的HP75细胞随机分为沉默组和阴性对照组，沉默组转染shRNA-FGD5-AS1，阴性对照组转染shRNA-NC,RT-PCR检测2组细胞中FGD5-AS1和miR-15a的表达水平，CCK-8实验、细胞划痕实验、Transwell实验检测2组细胞增殖、迁移及侵袭能力，Western blot检测2组细胞Wnt/β-catenin信号通路相关蛋白表达，双荧光素酶报告基因实验验证FGD5-AS1和miR-15a的靶向关系。结果 与NHA细胞相比，HPAs、RC-4BC、HP75细胞中FGD5-AS1的表达水平显著升高（P<0.05），而miR-15a的表达水平显著降低（P<0.05）。与阴性对照组相比，沉默组HP75细胞中的FGD5-AS1表达水平降低（P<0.05）,miR-15a表达水平升高（P<0.05）,OD值下降（P<0.05），细胞迁移及侵袭能力降低（P<0.05）,Wnt3a和β-catenin蛋白表达降低（P<0.05）。FGD5-AS1能特异性结合miR-15a使细胞荧光素酶活性降低（P<0.05）。结论 FGD5-AS1在PA细胞中表达升高，沉默FGD5-AS1能够抑制PA细胞的增殖、迁移及侵袭，其机制与其对miR-15a的靶向调控有关。更多还原

【Abstract】 Objective To investigate the effects of long non-coding RNA FGD5-AS1 on the proliferation,migration,and invasion of pituitary adenoma(PA) cells,and to analyze its potential mechanism of action.Methods Human PA cell lines HPAs,RC-4 BC,HP75,and human astrocyte cell line NHA were cultured in vitro. The expression levels of FGD5-AS1 and miR-15a in the above cell lines were detected by RT-PCR. HP75 cells in the logarithmic growth phase were randomly divided into the silencing group and the negative control group. The silencing group was transfected with shRNA-FGD5-AS1,while the negative control group was transfected with shRNA-NC. The expression levels of FGD5-AS1 and miR-15a in the two groups of cells were detected by RT-PCR. The proliferation,migration and invasion abilities of the two groups of cells were determined by CCK-8 assay,wound healing assay,and Transwell assay. The expression of proteins related to the Wnt/β-catenin signaling pathway in the two groups of cells was detected by Western blot. The targeting relationship between FGD5-AS1 and miR-15a was verified by dual-luciferase reporter gene assay.Results Compared with the NHA cell,the expression level of FGD5-AS1 was significantly increased in the HPAs,RC-4 BC,and HP75 cells((P<0. 05),whereas the expression level of miR-15a was significantly decreased(P<0. 05). Compared with the negative control group,the expression level of FGD5-AS1 was decreased(P<0. 05),the expression level of miR-15a was increased(P<0. 05),the OD value was decreased(P<0. 05),the migration and invasion abilities of cells were reduced(P<0. 05),and the expression of Wnt3a and β-catenin proteins was decreased in the silencing group of HP75 cells(P<0. 05). FGD5-AS1 could specifically bind to miR-15a,leading to a decrease in cell luciferase activity(P<0. 05).Conclusion FGD5-AS1 is overexpressed in PA cells,and silencing FGD5-AS1 can inhibit the proliferation,migration,and invasion of PA cells,and the mechanism is related to its targeted regulation of miR-15a.更多还原