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目的 探讨nm2 3 H1基因转染和forskolin对人高转移大细胞肺癌细胞株L9981PKA活性的影响。方法 应用PKA通路特异激动剂forskolin对人高转移大细胞肺癌原代细胞株L9981、转基因细胞株L9981 nm 2 3 H1 pLXSN和空载体转染细胞株L9981 pLXSN共同培养 ,应用放免法检测forskolin处理后不同时间点三个细胞株PKA的活性变化。结果 ( 1)forskolin处理前L9981、L9981 pLXSN和L9981 nm2 3 H1 pLXSN的PKA活性经F检验有显著性差异 (P <0 .0 1) ;两两比较 :L9981 nm2 3 H1 pLXSN的PKA活性显著高于L9981和L9981 pLXSN(P <0 .0 1) ,而L9981与L9981 pLXSN之间比较无显著性差异 (P >0 .0 5 )。 ( 2 )在同一作用时间 ( 3 0min)下 ,三个细胞株经不同浓度forskolin处理后PKA活性均显著升高 (P <0 .0 1) ,在forskolin浓度为 10 0 μmol/L时PKA活性最高 ,呈剂量依赖关系。 ( 3 )三个细胞株在同一浓度forskolin( 10 0μmol/L)处理下 ,PKA活性随作用时间升高 ,在forskolin作用时间为 3 0min时PKA的活性最高 ,呈时间依赖关系。结论 ( 1)nm2 3 H1基因转染能显著上调人高转移大细胞肺癌细胞株L9981的PKA活性 ,nm2 3 H1作为一种肿瘤转移抑制基因的作用机理与PKA信号通路可能有一定联系 ;( 2 )forskolin能显著上调L9981细胞?
Objective To investigate the effects of nm23 H1 gene transfection and forskolin on the activity of L9981PKA in human high metastatic large cell lung cancer cell line L9981. Methods Forskolin, a PKA pathway-specific agonist, was used to co-culture primary human lung cancer cell line L9981, L9981 nm 2 3 H1 pLXSN and L9981 pLXSN. The expression of forskolin was detected by radioimmunoassay Changes in PKA activity of three cell lines at time points. Results (1) The PKA activities of L9981, L9981 pLXSN and L9981 nm2 3 H1 pLXSN before forskolin treatment were significantly different (P <0.01) by F test. The pairwise comparison showed that the PKA activity of L9981 nm2 3 H1 pLXSN was significantly higher There was no significant difference between L9981 and L9981 pLXSN (P <0.01), but no significant difference between L9981 and L9981 pLXSN (P> 0.05). (2) Under the same action time (30min), the PKA activity of three cell lines were significantly increased after treatment with different concentration of forskolin (P <0.01), PKA activity at forskolin concentration of 10μmol / L Highest, in a dose-dependent manner. (3) Under the same concentration of forskolin (10 0μmol / L), the PKA activity of three cell lines increased with time, and the activity of PKA was the highest when the concentration of forskolin was 30min, in a time-dependent manner. CONCLUSION: (1) The transfection of nm23 H1 gene can significantly up-regulate the PKA activity in human high metastatic large cell lung cancer cell line L9981. The possible mechanism of nm23 H1 as a tumor metastasis suppressor gene may be related to the PKA signaling pathway. ) forskolin can significantly up-regulate L9981 cells?