下调磷酸甘油酸激酶1增强胶质瘤 U251细胞的放射敏感性

摘要

Objective To explore the role of phosphoglycerate kinase 1 ( PGK1 ) in the radiosensitivity of glioma U251 cells.Methods Radioresistant U251 cells ( RR-U251 ) were established .The shRNA for PGK1 and the control shRNA were transfected to U 251 cells using lipofectamineTM2000, and real-time quantitative fluorescent PCR and western blot analysis were performed to evaluate PGK1 expression .The cell viability was determined by MTT assay .The wound-healing assay ( WHA) method was used to evaluate cell migration ability .Cell invasion abilities were examined using Transwell culture chamber system .Results Our results showed that the expression of PGK1 was significantly increased in RR-U251 cells compared with normal U251 cells.After irradiation , the cell viability was significantly higher in RR-U251 cells compared with normal U 251 cells, and migration ability and invasion ability was also enhanced .Down-regulating PGK1 using shRNA could induce a significantly down-regulated cell viability ,and decreased the ability of migration and invasion.Conclusion These findings suggest that down-regulation of PGK1 could enhance radiosensitivity in human U 251 cells,and PGK1 may serves as a target for enhancing radiosensitivity .%目的：研究磷酸甘油酸激酶1（ PGK1）对胶质瘤U251细胞放射敏感性的影响。方法建立放射抵抗的U251（RR-U251）细胞；LipofectamineTM2000方法将抑制PGK1表达的shRNA-PGK1质粒或对照shRNA-NC转染至U251细胞；荧光定量PCR及Western Blot检测PGK1的表达；MTT检测细胞相对存活率，划痕实验检测细胞迁移能力，Transwell小室实验检测细胞侵袭能力。结果与正常U251细胞相比，RR-U251细胞中PGK1的表达明显增高；放射处理后，RR-U251细胞的相对存活率升高，迁移能力及侵袭能力增强。转染shRNA-PGK1的细胞接受放射处理后，与对照组相比，细胞的相对存活率降低，迁移能力以及侵袭能力减弱。结论下调U251细胞中PGK1的表达可增加其放射敏感性，提示PGK1可能成为增强放射敏感性的调控靶点。