蛋白激酶C信号通路对温石棉介导的肺成rn纤维细胞细胞周期调控蛋白表达的影响

摘要

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)Cyclin D1、Cyclin E、增殖细胞核抗原(PCNA)和Cyclin A表 达阳性率分别为19.0%、22.8%、79.7%和65.1%，均较各对照组低，差异有显著性(P<0.0 5)，且Cyclin D1和Cyclin E表达阳性率低于PCNA和Cyclin A，差异有显著性(P<0.01 )；而Cyclin B1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<0.05)，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，其中PCNA表达的改变可 能与上游PKC信号通路有关。%Objective　To study the relation of cell cycle-regulating protein expression with up-stream protein kinase C(PKC) signa l 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(SiO2),nega tive control(TiO2),normal control(rabbit alveolar macrophage,AM) and blank con trol were set up. Results　In chrysotile group,the p ositive expression rate of Cyclin D1,Cyclin E,PCNA and Cyclin A was 19.0%,22.8 %,79.7% and 65.1% respectively,significantly lower than that of the differ ent controls(P<0.05),the positive expression rate of Cyclin D1 and Cyclin E was significantly lower than that of PCNA and Cyclin A(P<0.01).But the pos itive expression rate of p34cdc2 kinase and Cyclin B1 was higher than that of B.control and AM group. When PKC signal pathway was inhibited by PKC inhibitor,t he positive expression rate of PCNA significantly decreased(P<0.05) in chrys otile 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.