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目的 探讨转移抑制基因nm2 3 H1对人高转移大细胞肺癌细胞株L9981中Wnt信号传导激酶GSK 3 β的表达量和活性的影响 ,为全面阐明nm2 3 H1基因调控肺癌转移相关信号传导通路的分子机制提供实验依据。方法 将稳定转染nm 2 3 H1基因的人高转移大细胞肺癌细胞株L9981 nm2 3 H1、原代细胞株L9981和空载体转染细胞株L9981 pLXSN培养传代 ,应用Westernblot分别检测应用 2 0mmol/LLiCl处理前后三个肺癌细胞株胞浆、胞核中GSK 3 β的表达水平 ,应用免疫共沉淀同位素闪烁计数法测定上述肺癌细胞株胞浆和胞核中的GSK 3 β激酶活性。 结果 ( 1)L9981 nm2 3 H1胞浆和胞核中GSK 3 β蛋白表达量分别为( 63 41± 5 41)和 ( 4 3 5 6± 490 )IOD ,L9981 pLXSN分别为 ( 3 613± 3 83 )和 ( 70 5± 75 )IOD ,L9981分别为 ( 373 6± 2 98)和 ( 65 7± 5 7)IOD。三个细胞株间胞浆和胞核中GSK 3 β表达量均有非常显著差异 (P <0 .0 1) ;两两比较 :L9981 nm2 3 H1胞浆和胞核GSK 3 β表达水平均显著高于L9981 pLXSN和L9981(P <0 .0 1) ,而后两者之间比较均无显著性差异 (P >0 .0 5 )。 ( 2 )L9981 nm2 3 H1胞浆和胞核GSK 3 β激酶活性分别为 ( 2 895 5±2 5 0 9)和 ( 92 47± 92 4)CPM ,L9981 pLXSN分别为 ( 112 41± 1
Objective To investigate the effect of metastasis suppressor gene nm23 H1 on the expression and activity of Wnt signaling kinase GSK 3 β in human high metastatic large cell lung cancer cell line L9981. In order to fully elucidate the molecular mechanism of nm23 H1 gene regulating lung metastasis-associated signal transduction pathway Mechanism to provide experimental basis. Methods Human high metastatic large cell lung cancer cell line L9981 nm23 H1, primary cell line L9981 and empty vector transfected cell line L9981 pLXSN stably transfected with nm 2 3 H1 gene were passaged and passaged. Western blot was used to detect the expression of 20 mmol / L LiCl The levels of GSK 3 β in cytoplasm and nucleus of three lung cancer cell lines were measured before and after treatment. The activity of GSK 3 β-kinase in cytoplasm and nucleus of lung cancer cell lines was determined by co-immunoprecipitation isotope scintillation counting. Results (1) The expressions of GSK 3 β protein in cytoplasm and nucleus of L9981 nm2 3 H1 cells were (63 41 ± 5 41) and (4356 ± 490) IOD, respectively. The L9981 pLXSN were (3 613 ± 3 83 ) And (70 5 ± 75) IOD and L9981 were (373 6 ± 2 98) and (65 7 ± 5 7) IOD, respectively. The expression of GSK 3 β in the cytoplasm and the nucleus of the three cell lines was significantly different (P <0.01). The pairwise comparison showed that the expression of GSK 3 β in cytoplasm and nucleus of L9981 nm 2 3 H1 cells were significantly Higher than that of L9981 pLXSN and L9981 (P <0.01), but there was no significant difference between the two groups (P> 0.05). (2) The cytoplasmic and nuclear GSK 3 β-kinase activities of L9981 nm2 3 H1 cells were (2 895 5 ± 2 5 0 9) and (92 47 ± 92 4) CPM, respectively. L9981 pLXSN was (112 41 ± 1)