nm23-H1基因靶向调控人肺癌细胞PKA信号通路的实验研究

An experimental study on the targeted regulation effect of the nm23-H1 gene for PKA signal pathway of lung cancer cell

目的探讨nm-23H1基因靶向调控人高转移大细胞肺癌细胞株L9981PKA信号通路的分子机理。方法应用放免法、Westem blot法检测人高转移大细胞肺癌细胞株L9981、空载体转染细胞株L9981-pLXSN和转基因细胞株L9981-nm23-H1中的PKA活性、CREB和p-CREB的表达水平。结果L9981-nm-23H1肺癌细胞株PKA活性[1.906±0.034μmol/(min.g)]显著高于L9981[0.441±0.021μmol/(min.g)]和L9981-pLXSN[0.443±0.059μmol/(min.g)]肺癌细胞株(P=0.000);而L9981与L9981-pLXSN肺癌细胞株间PKA活性比较无显著性差异(P=0.991);(2)L9981、L9981-pLXSN和L9981-nm23-H1肺癌细胞株间CREB表达水平比较无显著性差异(P=0.774);L9981-nm23-H1肺癌细胞株p-CREB表达量显著高于L9981和L9981-pLXSN肺癌细胞株(P=0.000),而L9981与L9981-pLXSN肺癌细胞株间p-CREB表达量比较无显著性差异(P=0.126)。结论(1)nm23-H1基因转染能显著上调人高转移大细胞肺癌细胞株L9981PKA信号通路关键激酶的活性和表达量;(2)nm23-H1基因逆转人高转移大细胞肺癌侵袭转移表型的分子机理可能与其激活PKA信号传导有关。

Objective To investigate the molecular mechanism of nm23 - H 1 gene on the targeted regulation effect for PKA signal pathway of lung cancer cell. Methods The PKA activity, CREB and p - CREB expression levels among primary human high- metastatic large cell lung cancer cell line （L9981）, vector transfection lung cancer cell line （L9981 - pLXSN） and nm23- H1 gene transfection lung cancer cell line （L9981 -nm23- H1） by radioimmunoassay and Western blot methods respectively. Results （1） The activity of PKA in L9981 - nm23 - H1 lung cancer cell line [ 1. 906 ± 0. 034μmol/ （min·g） ] was significantly higher than that in L9981 [ 0. 441 ± 0.021μmol/ （min·g）] and L9981 - pLXSN lung cancer cell lines [0.443 ± 0.059μmol/ （min·g）] （P = 0. 000）, but no significant difference of the PKA activity was observed between L9981 and L9981 -pLXSN lung cancer cell lines （P 〉 0.05） ; （2） No significant difference of the expression level of CREB was observed among L9981, L9981 -pLXSN and L9981 - nm23 -H1 lung cancer cell lines （P 〉 0.05）, but the expression level of p - CREB in L9981 - nm23 - H1 lung cancer cell line was remarkably higher that that in L9981 and L9981 - pLXSN lung cancer cell lines （ P = 0. 000）. Conclusion （ 1 ） Transfection of nm23 - H1 gene can significantly up -regulate the activity and expression related invasion and metastasis of human high- metastatic large cell lung cancer might be related to its effecfs of activating PKA signal pathway.