nm23-H_1基因转染前后人肺癌细胞中PKC转位的研究  

Study on protein kinase C translocation before and after transfection of nm23-H1 gene in human lung cancer cells using Laser scanning confocal microscope

作　　者：聂强[1] 朱文[1] 王艳萍[1] 陈小禾[1] 杨俊杰[1] 刘伦旭[1] 付军科[1] 李定彪[1] 李印[1] 周清华[1] 

机构地区：[1]四川大学华西医院四川省肺癌分子重点实验室,胸外科,成都610041

出　　处：《中国肺癌杂志》2004年第2期86-90,共5页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金 (No .30 0 70 333和No.30 1 0 0 0 75)资助~~

摘　　要：目的　探讨nm2 3 H1转染和蛋白激酶C(PKC)特异抑制剂CalphostinC对人高转移大细胞肺癌细胞株L9981细胞PKC信号转导通路的作用 ,以及nm2 3 H1基因对PKC激活转位的影响。方法　应用激光扫描共聚焦显微镜观察nm 2 3 H1基因转染前后和CalphostinC处理转基因细胞株L9981 nm2 3 H1前后PKC在不同的亚细胞区域的定位情况。结果　 ( 1)原代细胞株L9981和空载体细胞株L9981 pLXSN中PKC α、PKC βⅡ主要位于胞核及核周 ,处于激活状态 ;转染nm 2 3 H1基因后的人肺癌细胞株L9981 nm2 3 H1中PKC α、PKC βⅡ主要位于胞浆 ,处于未激活状态。 ( 2 )CalphostinC作用后所有细胞中的PKC均主要位于胞浆中 ,处于未激活状态。结论　 ( 1)nm2 3 H1基因可使L9981细胞株中PKC从胞核向胞浆转位 ,从而抑制PKC信号转导。 ( 2 )CalphostinC可使L9981、L9981 pLXSN细胞株中PKC从胞核向胞浆转位 ,从而抑制PKC信号转导。Objective To explore the influences of nm23-H1 gene transfection and protein kinase C (PKC) inhibitor Calphostin C on PKC signal transduction pathway in human high-metastasis large cell lung cancer cell line L9981, and to evaluate the effects of nm23-H1 gene on translocation and activation in subcellular region. Methods The translocation of PKC in subcellular region was observed in L9981 before and after nm23-H1 gene transfection and Calphostin C treatment by Laser scanning confocal microscope (LSCM) method. Results PKC-α and PKC-βⅡ were found to locate in different subcellular site in L9981 before and after nm23-H1 gene transfection. PKC-α and PKC-βⅡ mainly located in nucleus and perinucleus in L9981 and L9981-pLXSN cell lines, which were in active status. PKC-α and PKC-βⅡ mainly located in soluble cytosolic fraction in L9981-nm23-H1 cell line and were inactive status. PKC-α and PKC-βⅡ mainly located in cytosolic fraction and were in inactive status in all the three cell lines after treatment with Calphostin C. Conclusion The results suggest that nm23-H1 gene might make PKC to translocate from nucleus and perinucleus to soluble cytosolic fraction in L9981 cell line. PKC inhibitor, Calphostin C, can also make PKC to translocate from nucleus and perinucleus to soluble cytosolic fraction in L9981, L9981-pLXSN cell lines. Both transfection of nm23-H1 gene and treatment with Calphostin C can suppress the PKC signal transduction in L9981 cell line.

关 键 词：NM23-H1 基因转染 肺癌 癌细胞 PKC转位 蛋白激酶C抑制剂 信号转导 

分 类 号：R734.2[医药卫生—肿瘤] R394[医药卫生—临床医学]