蛋白激酶C对血管内皮细胞锚蛋白、CD44表达及亚细胞分布的影响

Effects of Protein Kinase C on Expression and Subcellular Distribution of Ankyrin and CD44 in Vascular Endothelial Cells

通过外源性底物对[γ-32P]-ATP的摄入量来测定豆蔻酰佛波醇乙酯(phorbol-myristate-acetate,PMA)处理后的人脐静脉内皮细胞(humanumbilicalveinendothelialcells,HUVECs)膜蛋白激酶C(proteinkinaseC,PKC)的活性;利用间接免疫荧光标记和Western印迹方法分析蛋白激酶C活性对锚蛋白及CD44的亚细胞分布及蛋白质表达的影响。结果发现HUVECs的锚蛋白及CD44表达水平趋势与PKC活性变化相吻合;PKC活化导致CD44在细胞膜上呈聚集状,而锚蛋白则移位并聚集于CD44处;PKC抑制剂能抑制PKC活化所带来的上述作用。结果表明PKC活化通过磷酸化作用能上调锚蛋白及CD44表达,并同时导致二者发生一致性运动及共分布。

Membranous protein kinase C （PKC） activity after phorbol-myristate-acetate （PMA） treatment was determined by the incorporate quantity of [γ-32P]-ATP into exogenous substrate in human umbilical vein endothelial cells （HUVECs）. Effects of PKC activity on subcellular distribution and protein expression of ankyrin and CD44 was analyzed respectively with indirect immunofluorescence and Western blot. We found that the trend of ankyrin and CD44 expression was identical with PKC activity change. PKC activation could result in CD44 aggregating on HUVECs membrane, and ankyrin also translocated and aggregated beneath CD44. Calphostin C could inhibit the above action of PKC. The results showed that PKC activation could up-regulate protein expression of ankyrin and CD44 by phosphorylation, and resulted in the coherent migration and colocalization of ankyrin and CD44 simultaneously.