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目的鞘氨醇激酶1(sphingosine kinase 1,SK1)具有调控细胞迁移的重要作用,丝/苏氨酸激酶(serine/threonine kinase,AKT)参与其调控过程。本研究旨在探讨SK1对结肠癌细胞株SW480迁移和侵袭的影响及与AKT信号通路的关系。方法以不同浓度12-豆蔻酸-13-乙酸佛波醇(phorbol 12-myristate-13-acetate,PMA)干预SW480细胞,观察细胞SK1、p-AKT基因和蛋白表达水平变化情况。采用Ttranswell法观察PMA和AKT抑制剂对SW480细胞迁移和侵袭能力的影响。结果 CCK8实验显示,25、50、100nmol/L PMA刺激的SW480细胞培养液吸光度(A)值分别为0.780±0.030、0.844±0.057和0.998±0.045,与空白对照组(A值0.631±0.051)比较均明显增加,F=93.48,P<0.001;PMA可诱导细胞内SK1基因和蛋白表达水平明显增高;Transwell实验结果显示,100nmol/L PMA增强SW480细胞迁移和侵袭(分别为169.8±10.82和89.6±8.44),较对照组(分别为63.6±8.67和39.6±6.12)明显增加,而AKT抑制剂AKTi1/2可减少PMA诱导的迁移和侵袭SW480细胞数量(分别为91.8±9.36和57.2±8.98);PMA增强SW480细胞SK1表达同时促进AKT蛋白磷酸化水平,而AKTi1/2可拮抗PMA的作用。结论 SK1可促进SW480细胞的迁移和侵袭能力,其机制与上调AKT信号通路有关。
Purpose Sphingosine kinase 1 (SK1) plays an important role in the regulation of cell migration. Serine / threonine kinase (AKT) is involved in its regulation. This study aimed to investigate the effect of SK1 on the migration and invasion of colon cancer cell line SW480 and its relationship with AKT signaling pathway. Methods SW480 cells were treated with different concentrations of phorbol 12-myristate-13-acetate (PMA) to observe the changes of SK1 and p-AKT gene and protein expression. Ttranswell method was used to observe the effects of PMA and AKT inhibitor on the migration and invasion ability of SW480 cells. Results The CCK8 assay showed that the absorbency (A) values of SW480 cells stimulated with 25, 50 and 100 nmol / L PMA were 0.780 ± 0.030, 0.844 ± 0.057 and 0.998 ± 0.045, respectively, compared with the blank control group (A value 0.631 ± 0.051) PMA could induce the expression of SK1 gene and protein in cells significantly increased. Transwell assay showed that 100 nmol / L PMA enhanced the migration and invasion of SW480 cells (169.8 ± 10.82 and 89.6 ± 8.44) compared with the control group (63.6 ± 8.67 and 39.6 ± 6.12 respectively), while the AKT inhibitor AKTi1 / 2 decreased the number of PMA-induced migration and invasion of SW480 cells (91.8 ± 9.36 and 57.2 ± 8.98, respectively); PMA enhanced the expression of SK1 in SW480 cells and promoted the phosphorylation of AKT protein, while AKTi1 / 2 antagonized the effect of PMA. Conclusion SK1 can promote the migration and invasion of SW480 cells. The mechanism is related to the up-regulation of AKT signaling pathway.