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为探讨巨噬细胞集落刺激因子(M-CSF)对人宫颈癌细胞(He La细胞)侵袭和迁移的影响及机制,采用胞质定位空载体p CMV/cyto/myc与重组载体p CMV/cyto/myc-M-CSF稳定转染He La细胞株,建立稳定高表达胞质M-CSF的细胞系(He La-M细胞).经Transwell实验观察胞质M-CSF对He La细胞侵袭和迁移能力的影响,逆转录-聚合酶链式反应及蛋白质印迹检测细胞Rho三磷酸鸟苷酶(Rho GTPases)及基质金属酶的表达,明胶酶谱法检测基质金属蛋白酶2的活性.结果显示,与转染空载体的He La细胞(He La-C细胞)和对照组He La细胞比较,胞质M-CSF的高表达可明显增强He La细胞在体外的侵袭和迁移能力,其机制与Rho GTPases的活化,以及MMP2表达上调及其活性增高密切相关.
To investigate the effect and mechanism of macrophage colony-stimulating factor (M-CSF) on invasion and migration of human cervical cancer cells (HeLa cells), we used cytoplasmic empty vector p CMV / cyto / myc and recombinant vector p CMV / cyto / He-M-CSF were transfected into HeLa cells to establish He La-M cells stably expressing Cytosolic M-CSF.The invasion and migration of HeLa cells were observed by Transwell assay (Rho GTPase) and matrix metalloproteinase (Rho GTPases) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, and the activity of matrix metalloproteinase 2 was detected by gelatin zymography.The results showed that He La cells (He La-C cells) transfected with empty vector Hela cells compared with the control group, the high expression of cytosolic M-CSF can significantly enhance the He La cells in vitro invasion and migration, the mechanism and Rho GTPases Activation, as well as MMP2 expression and its activity is closely related.