首页|SRSF9/SRp30c通过调控GRβ对胶质瘤细胞增殖和迁移的影响

SRSF9/SRp30c通过调控GRβ对胶质瘤细胞增殖和迁移的影响

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目的:探讨富含丝氨酸-精氨酸剪切因子9/富含丝氨酸-精氨酸蛋白30c(SRSF9/SRp30c)和糖皮质激素受体β(GRβ)在胶质瘤细胞中的表达及两者之间的关系.方法:用小干扰RNA(siRNA)靶向干扰胶质瘤U87细胞中SRSF9的表达,同时用慢病毒将短发卡RNA(shRNA)转染细胞,构建SRSF9敲减的U87稳转细胞株,通过荧光显微镜观察并检测转染情况.通过RT-qPCR和Western blot法检测SRSF9/SRp30c和GRβ的表达水平,CCK-8和细胞集落形成实验检测细胞的增殖能力,划痕实验检测细胞的迁移能力.结果:用2种方法敲减SRSF9后,SRSF9/SRp30c表达均降低,GRβ的表达水平也随之下降,差异有统计学意义(P<0.05).荧光显微镜观察结果表明,成功构建了稳转细胞株,转染效率超过80%.CCK-8实验和细胞集落形成实验的结果表明U87细胞在敲减SRSF9后,细胞活力和集落形成能力降低,差异有统计学意义(P<0.05);划痕实验结果表明U87细胞在敲减SRSF9后,迁移能力明显减弱,差异有统计学意义(P<0.05).结论:SRSF9/SRp30c可通过调控GRβ促进胶质瘤细胞的增殖和迁移.
Effects of SRSF9/SRp30c on proliferation and migration abilities of glio-ma cells by regulating GRβ
AIM:To investigate the expression of serine-arginine-rich splicing factor 9/serine-arginine-rich protein 30c (SRSF9/SRp30c) and glucocorticoid receptor β(GRβ) in the glioma cells and the relationship of them. METHODS:Small interfering RNA ( siRNA) was used to knock down the expression of SRSF9 in the U87 cells.Short hairpin RNA ( shRNA) derived from lentivirus was used to establish U 87 stable knockdown cell line .Fluorescence micros-copy was used to observe and detect transfection efficiency .The expression of Grβand SRSF9/SRp30c at mRNA and pro-tein levels was determined by RT-qPCR and Western blot .The cell viability , colony formation ability and migration ability were measured by CCK-8 assay, colony formation assay and wound healing experiment .RESULTS:The mRNA and pro-tein levels of SRSF9/SRp30c and Grβin the U87 cells were both down-regulated after knockdown of SRSF9 (P<0.05). Fluorescence microscopic observation showed that a stable cell line was constructed successfully , and the transfection effi-ciency exceeded 80%.After knockdown of SRSF9 expression in the U87 cells, the cell viability and colony formation abili-ty were reduced (P<0.05).The migration ability was weakened significantly after SRSF9 was knocked down (P<0.05). CONCLUSION:SRSF9/SRp30c may promote the proliferation and migration of the glioma cells by regulating GRβ.

GliomaGlucocorticoid receptor βSerine-arginine-rich splicing factor 9/serine-arginine-rich protein 30cCell proliferationCell migration

刘艳、邵阿末、殷莹、李正、张锐、宗达、邹健、胡亚玲

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无锡卫生高等职业技术学校,江苏无锡214028

无锡人民医院中心实验室,江苏无锡214023

胶质瘤 糖皮质激素受体β 富含丝氨酸-精氨酸剪切因子9/富含丝氨酸-精氨酸蛋白30c 细胞增殖 细胞迁移

国家自然科学基金资助项目

81372710

2017

中国病理生理杂志
中国病理生理学会

中国病理生理杂志

CSTPCDCSCD北大核心
影响因子:1.065
ISSN:1000-4718
年,卷(期):2017.33(10)
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