骨桥蛋白通过激活AKT信号通路促进肝癌细胞的侵袭转移

Osteopontin stimulates hepatoma cells invasion and metastasis through alanine aminotransferase assay signaling pathway

目的观察骨桥蛋白(Osteopontin,OPN)对AKT信号通路及肝癌细胞生物学行为的影响。方法人肝癌细胞系Hep G2细胞分别转染对照质粒pCDNA3.1和重组质粒pCDNA3.1-OPN,Western blot检测细胞中OPN、AKT、AKT-P、MMP2和uPA的表达水平,CyQUANT法和Matrigel侵袭实验观察OPN上调对细胞粘附和细胞侵袭的影响。结果 OPN激活AKT信号通路,与转染对照质粒的Hep G2细胞相比,转染OPN重组质粒的细胞MMP2和uPA表达增加,其细胞粘附百分比是56.08±2.0,大于对照组的49.93±1.74(P<0.05);穿过基底膜细胞数目是(25.2±2.08)个,大于对照组的(17.4±1.45)个(P<0.05)。结论 OPN可通过激活AKT信号通路,上调MMP2和uPA表达,促进Hep G2细胞侵袭转移。

Objective To study the effects of up-regulation of osteopontin（OPN） on AKT signaling pathway and biological behaviors of Hep G2 cells. Methods The recombinant vector pCDNA3.1-OPN and the control vector pCDNA3.1 were transferred into Hep G2 cells. The expression of OPN, AKT, AKT-P, MMP2 and uPA were detected by Western blot. The malignant phenotypes of transfected Hep G2 cells including adhesion and invasive activities were analyzed. Results OPN induced the AKT signaling pathway and enhanced the expression of MMP2 and uPA. Both the percentage of adhesion （56.08 ± 2.0） and the migrating number （25.2 ± 2.08）of Hep G2 cells transfected with pCDNA3. 1-OPN was significantly increased as compared with the controls （49.93 ± 1.74,17.4 ± 1.45） （P 〈0.05）. Conclusion OPN can induce the AKT signaling pathway, up-regulate the expression of MMP2 and uPA and stimulate the invasion and metastasis of Hep G2 cells.