蛋白激酶C信号通路对温石棉介导的肺成纤维细胞细胞周期调控蛋白表达的影响

Influence of protein kinase C signal pathways on the expression of cell cycle regulating protein in chrysotile induced proliferation of human embryonic pulmonary fibroblasts

目的 研究细胞周期调控蛋白在温石棉所致人胚肺成纤维细胞 (HEPF)增殖中的表达和蛋白激酶C(PKC)对细胞周期调控蛋白的影响。方法 利用体外细胞培养技术 ,用流式细胞仪测定温石棉介导的HEPF的细胞周期调控蛋白的表达 ,同时设立阳性对照 (标准SiO2 )、阴性对照 (标准TiO2 )、正常对照 (未经粉尘处理的兔肺泡巨噬细胞 )和空白对照。结果 温石棉组细胞周期蛋白 (cyclin)CyclinD1、CyclinE、增殖细胞核抗原 (PCNA)和CyclinA表达阳性率分别为 19.0 %、2 2 .8%、79.7%和 6 5 .1% ,均较各对照组低 ,差异有显著性 (P <0 .0 5 ) ,且CyclinD1和CyclinE表达阳性率低于PCNA和CyclinA ,差异有显著性 (P <0 .0 1) ;而CyclinB1和p34cdc2激酶的阳性表达率较空白对照和正常对照略高。当PKC信号通路被阻断后 ,PCNA在温石棉组表达的阳性率明显降低 ,差异有显著性 (P <0 .0 5 ) ,p34cdc2激酶的阳性表达率改变不明显。结论 这几种蛋白可能均参与了温石棉所致HEPF增殖 。

Objective To study the relation of cell cycle regulating protein expression with up stream protein kinase C(PKC) signal transduction pathways and the effect of PKC chrysotile induced proliferation of human embryonic pulmonary fibroblast(HEPF). Methods Flow cytometry to measure the expression of cell cycle regulating protein of HEPF mediated by chrysotile in vitro was used,and positive control(SiO 2),negative control(TiO 2),normal control(rabbit alveolar macrophage,AM) and blank control were set up. Results In chrysotile group,the positive expression rate of Cyclin D 1,Cyclin E,PCNA and Cyclin A was 19.0%,22.8%,79.7% and 65.1% respectively,significantly lower than that of the different controls( P <0.05),the positive expression rate of Cyclin D 1 and Cyclin E was significantly lower than that of PCNA and Cyclin A( P <0.01).But the positive expression rate of p34cdc2 kinase and Cyclin B 1 was higher than that of B.control and AM group. When PKC signal pathway was inhibited by PKC inhibitor,the positive expression rate of PCNA significantly decreased( P <0.05) in chrysotile group,and the change in positive expression rate of p34cdc2 kinase was not obvious.Conclusion These cell cycle regulating proteins may be involved in chrysotile induced proliferation of HEPF.The changes of PCNA expression may relate to the up stream PKC signal pathway.