nm23-H_1基因转染对人高转移大细胞肺癌细胞株L9981中β-catenin表达的影响

Effects of transfection of nm23-H1 gene on β-catenin expression in human high-metastatic large cell lung cancer cell line L9981

目的 探讨nm2 3 H1基因转染对人高转移大细胞肺癌细胞株L9981中 β catenin和磷酸化β catenin表达水平的影响 ,为阐明nm2 3 H1基因逆转肺癌转移的分子机制提供实验依据。方法 以原代L9981、转染nm 2 3 H1基因的L9981 nm2 3 H1和转染空载体的L9981 pLXSN三株肺癌细胞株为研究对象 ,应用WesternBlot检测比较各细胞株胞浆、胞核中 β catenin和磷酸化 β catenin表达水平的变化 ,以确定nm 2 3 H1基因转染是否能调控人高转移大细胞肺癌细胞株L9981中 β catenin和磷酸化 β catenin表达。 结果 ①β catenin在L9981 nm2 3 H1细胞株胞浆中表达量 (IOD) (3 64 9± 118)显著高于L9981(14 0 1± 3 1)和L9981 pLXSN(13 5 0± 5 5 )细胞株 (P <0 .0 0 1) ;②β catenin在L9981 nm 2 3 H1细胞株胞核中表达量 (2 945± 68)与L9981(2 60 4± 2 3 )和L9981 pLXSN(2 65 2± 5 3 )细胞株比较均无显著性差异 (P >0 .0 5 ) ;③磷酸化β catenin在L9981 nm 2 3 H1细胞株胞浆中表达量 (3 12 3± 10 2 )显著低于L9981(4 3 62± 13 1)和L9981 pLXSN (4 5 0 0±117)细胞株 (P <0 .0 0 1) ;④磷酸化 β catenin在L9981 nm2 3 H1细胞株胞核中表达量 (5 13 6± 112 )显著高于L9981(2 666± 116)和L9981 pLXSN(2

Objective To explore the effects of transfection of nm23-H1 gene on expression of β-catenin and phospho-β-catenin in human high-metastatic large cell lung cancer line L9981, and to provide evidence to elucidate the molecular mechanism of nm23-H1 mediated tumor metastatic suppression. Methods To determine whether nm23-H1 contributes to cytoplasm and nuclear β-catenin and phospho-β-catenin expression, the expression level of β-catenin and phospho-β-catenin in cytoplasm and nucleus was detected in human high-metastatic large cell lung carcinoma cell lines including primary cell line L9981 with nm23-H1 gene deletion, L9981-nm23-H1 transfected with nm23-H1 gene, and L9981-pLXSN transfected with vector by Western blot. Results ①β-catenin expression in L9981-nm23-H1 cytoplasm (IOD) (3 649±118) was significantly higher than that in L9981 (1 401±31) and L9981-pLXSN (1 350±55) cell lines (P<0.001);②There was no statistical diffe-rence of the β-catenin expression in nucleus among L9981-nm23-H1 (2 945±68), L9981 (2 604±23) and L9981-pLXSN (2 652±53) cell lines (P>0.05);③Phospho-β-cetenin expression of cytoplasm in L9981-nm23-H1 cell line (3 123±102) was significantly lower than that in L9981 (4 362±131) and L9981-pLXSN ( 4 500 ±117) cell lines (P<0.001);④Phospho-β-catenin expression of nucleus in L9981-nm23-H1 (5 136±112) was significantly higher than that in L9981 (2 666±116) and L9981-pLXSN (2 661±66) cell lines (P<0.001);⑤There was no statistical difference of β-catenin or phospho-β-catenin expression in cytoplasm and nucleus between L9981 and L9981-pLXSN cell lines (P>0.05). Conclusion ①nm23-H1 gene transfection can remarkably upregulates the expression of cytoplasm β-catenin in human high-metastatic large cell lung cancer cell line L9981, but do not induce the nucleus accumulation of β-catenin; ②Transfection of nm23-H1 gene can significantly upregulate the expression of phospho-β-catenin in nucleus and remarkably downregulate the expression of phospho-β-catenin in cytoplasm of L9981; ③Regulation of the expression of the key modecule, β-catenin, in Wnt signal pathway might be the important melecular mechanisms which nm23-H1 gene controls “Lung Cancer Metastatic Suppresive Cascade” and reverves cancer metastasis in L9981.