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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 investigate the effect and mechanism of exogenous zinc ion on the activity of human respiratory epithelial cell tumor suppressor protein PTEN. Methods The human respiratory epithelial cell line BEAS-2B was used as an in vitro model. The malachite green assay was used to determine the effect of zinc ion on PTEN phosphatase activity. The effect of zinc ion on the phosphorylation of PTEN protein was detected by Western Blot. The effect of zinc ion on the activity of PTEN upstream kinase CK-2 was determined using casein kinase 2 (CK-2) kit. Results 50μmol / L zinc ion could inhibit PTEN phosphorylation and phosphatase activity (P <0.05). Under the same experimental conditions, zinc ion stimulation also inhibited the activity of CK-2 (P <0.05). In addition, CK-2 induced PTEN phosphorylation (P <0.05). Conclusion Exogenous zinc ion may further inhibit PTEN phosphatase activity in human respiratory epithelial cells by inhibiting the activity of CK-2.