ABSTRACT
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.
ABSTRACT
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.