目的观察细胞因子信号负调控因子3(suppressor of cytokine signaling3,SOCS3)对人肺腺癌细胞株A549侵袭能力及黏着斑激酶(focal adhesion kinase,FAK)磷酸化水平的影响。方法建立SOCS3稳定表达细胞株;Boyden Chamber分析法测定细胞的侵袭能力;RT-PCR检测FAK mRNA表达水平;Western blot检测FAK和磷酸化FAK表达水平。结果在纤维粘连蛋白(fibronectin,Fn)介导下,SOCS3加Fn组A549细胞穿过滤膜细胞数显著降低,差异有统计学意义(P〈0.01);A549细胞各组细胞间FAK mRNA水平差异无统计学意义;而SOCS3加Fn组细胞FAK蛋白及磷酸化FAK蛋白水平显著下降,差异有统计学意义(P〈0.05)。结论SOCS3从转录后水平调控FAK表达和活性从而降低A549的体外侵袭能力。
Objective To explore the effect of SOCS3 on invasion ability and FAK phosphorylation of human lung adenocarcinoma cell line A549. Methods A549 cell lines with SOCS3 stable expression were generated. Boyden Chamber assay was used to observe invasive ability of A549 cells. RT-PCR was used to detect FAK mRNA level. Western blot was used to detect the FAK protein and phosphorylated FAK protein level. Results Compared with the Fn group, the invasive abilitiy of A549 cells with SOCS3 stable expression was significantly lower (P〈0.01), the FAK protein and phosphorylated protein FAK level activated by fibronectin were inhibited significantly (P〈0.05 ,P〈0.01). However,there were no changes in FAK mRNA level in each A549 groups. Conclusion SOCS3 protein inhibits the invasive ability of A549 cells by downregulating FAK protein level and its activation at post-transcriptional level.