STIP1基因沉默对骨肉瘤细胞增殖、迁移和侵袭的影响

Effect of STIP1 Gene Silencing on Proliferation,Migration and Invasion of Osteosarcoma Cells

目的探讨磷酸化应激诱导蛋白1基因沉默(si-STIP1)对骨肉瘤细胞增殖、迁移和侵袭的影响。方法体外培养人成骨细胞(hFOB1.19细胞)和人骨肉瘤细胞系(U2OS、MG63和SaoS-2细胞),采用实时定量PCR和蛋白印迹法测定磷酸化应激诱导蛋白1(STIP1)在各细胞中的表达水平。si-STIP1及阴性对照(si-NC)转染U2OS细胞48 h,采用CCK-8法测定细胞增殖,Transwell实验测定细胞迁移和侵袭,采用蛋白印迹测定细胞MMP2和MMP9及IGF1R/PI3K/AKT信号通路蛋白的表达。结果与hFOB1.19细胞相比,U2OS、MG63和SaoS-2细胞的STIP1基因和蛋白表达水平均显著上调(P<0.05)。与si-NC组相比,si-STIP1组U2OS细胞的增殖活性、迁移率、侵袭率均显著降低(均P<0.05),MMP2、MMP9、p-IGF1R、p-PI3K和p-AKT蛋白表达水平均显著下调(P<0.05)。结论STIP1基因沉默抑制骨肉瘤细胞增殖、侵袭和迁移,其可能机制是通过调控IGF1R/PI3K/AKT信号通路实现。

Objective To study the effect of phosphorylated stress-induced phosphoprotein 1(STIP1)gene silencing on the proliferation,migration and invasion of osteosarcoma cells.MethodsHuman osteoblasts(hFOB1.19)and osteosarcoma cells(U2OS,MG63 and SaoS-2)were cultured,and the levels of STIP1 expression were detected by real-time quantitative PCR and protein blotting.Cell proliferation was determined by CCK-8 assay at 48 hours after U2OS cells were transfected with si-STIP1 and negative control(si-NC).Cell migration and invasion were evaluated by transwell assay.The expression of MMP2,MMP9 and IGF1R/PI3K/AKT signaling pathway proteins were measured by Western blot.Results Compared with hFOB1.19 cells,the expression of STIP1 mRNA and protein markedly increased in U2OS,MG63 and SaoS-2 cells(P<0.05).Compared with si-NC group,the proliferative activity,migration rate,invasion rate and expression of MMP2,MMP9,p-IGF1R,p-PI3K and p-AKT significantly decreased in si-STIP1 group(P<0.05).Conclusion STIP1 gene silencing can inhibit the proliferation,migration and invasion of osteosarcoma cells by regulating IGF1R/PI3K/AKT signaling pathway.