目的探讨外源性锌离子对人呼吸道上皮细胞肿瘤抑制蛋白PTEN的活性的影响及其机制。方法以人呼吸道上皮细胞株BEAS-2B作为体外模型,应用孔雀绿法测定锌离子对PTEN磷酸酶活性的影响;利用Western Blot法检测锌离子对PTEN蛋白磷酸化的影响。利用酪蛋白激酶2(CK-2)试剂盒测定锌离子对PTEN上游激酶CK-2活性的影响。结果 50μmol/L锌离子刺激可抑制PTEN蛋白磷酸化及其磷酸酶活性(P〈0.05)。在相同实验条件下,锌离子刺激还可抑制CK-2的活性(P〈0.05)。此外,CK-2可致PTEN磷酸化(P〈0.05)。结论外源性锌离子可能通过抑制CK-2的活性进一步抑制人呼吸道上皮细胞内肿瘤抑制蛋白PTEN磷酸酶活性。
Objective To examine the effect of exogenous zinc on enzymatic activity of tumor suppressor PTEN and underlying mechanisms.Methods Human airway epithelial cells(BEAS-2B) were employed as the in vitro model.Enzymatic activity of PTEN was measured using malachite green colorimetric assay and immunoblotting.Moreover,the activity of casein kinase-2(CK-2),the upstream kinase of PTEN,was determined.Results Exposure of BEAS-2B cells to Zn2+ inhibited phosphatase activity of PTEN,as well as PTEN phosphorylation,indicative of PTEN activity(P〈0.05).Under the same experimental condition,Zn2+ stimulation resulted in significant reduction of CK-2 activity(P〈0.05).In addition,CK-2 was shown to induce PTEN phosphorylation(P〈0.05).Conclusion These results suggested that Zn2+ exposure might inhibit PTEN phosphatase activity through inhibition of CK-2 activity in human airway epithelial cells.