Fu Junke, Zhou Qinghua, Zhu Wen, Wang Yanping, Liu Lunxu, Chen Xiaohe, Nie Qiang, Li Dingbiao, Li Yin
Key Laboratory of Lung Cancer Molecular Biology of Sichuan Province, and Department of Thoracocardiac Surgery, West China Hospital, Sichuan University , Chengdu, Sichuan 610041, P.R.China.
Zhongguo Fei Ai Za Zhi. 2004 Aug 20;7(4):294-7. doi: 10.3779/j.issn.1009-3419.2004.04.06.
To explore the possibility of targeting blockage of Wnt signal transduction pathway of nm23-H1 gene transfection in human high-metastatic large cell lung cancer cell line L9981, and to provide evidence to elucidate the signal conductive mechanism of nm23-H1 mediated tumor metastasis suppression.
The expression of GSK-3β and β-catenin of Wnt signal pathway was detected in cytoplasm and nucleus in L9981 cell line with nm23-H1 deletion, L9981-pLXSN cell line transfected with vector and L9981-nm23-H1 cell line transfected with nm23-H1 gene by Western blot.
(1)GSK-3β expression in L9981-nm23-H1 cytoplasm (6 341±541) was significantly higher than those in L9981 (3 736±298) and L9981-pLXSN (3 613±383) cell lines ( P < 0.001); (2)GSK-3β expression in L9981-nm23-H1 nucleus (4 356±490) was significantly higher than those in L9981 (657±57) and L9981-pLXSN (705±75) cell lines ( P < 0.001); (3)β-catenin expression in L9981-nm23-H1 cytoplasm (3 649±118) was significantly higher than those in L9981 (1 401±31) and L9981-pLXSN (1 350±55) cell lines ( P < 0.001); (4)No statistical difference of the β-catenin expression in nucleus was observed among L9981-nm23-H1 (2 945±68), L9981 (2 604±23) and L9981-pLXSN (2 652±53)( P > 0.05); (5)No significant difference of GSK-3β or β-catenin expression in cytoplasm and nucleus was observed between L9981 and L9981-pLXSN ( P > 0.05).
(1)nm23-H1 gene can remarkably upregulate the expression of GSK-3β in cytoplasm and nucleus, and β-catenin expression in cytoplasm in L9981-nm23-H1 cell, but can not induce the nucleus accumulation of β-catenin. (2)Regulation of GSK-3β and β-catenin expression, and targeting blockage of Wnt signaling pathway may be one of molecular mechanisms that nm23-H1 contributes to play a vital role in the "Lung Cancer Metastasis Suppressive Cascade".
探讨靶向阻断人高转移大细胞肺癌细胞系L9981中nm23-H1基因转染的Wnt信号转导通路的可能性，为阐明nm23-H1介导肿瘤转移抑制的信号传导机制提供依据。
采用蛋白质免疫印迹法检测nm23-H1基因缺失的L9981细胞系、转染空载体的L9981-pLXSN细胞系及转染nm23-H1基因的L9981-nm23-H1细胞系中Wnt信号通路中GSK-3β和β-连环蛋白在细胞质和细胞核中的表达。
（1）L9981-nm23-H1细胞质中GSK-3β表达量（6 341±541）明显高于L9981（3 736±298）和L9981-pLXSN（3 613±383）细胞系（P<0.001）；（2）L9981-nm23-H1细胞核中GSK-3β表达量（4 356±490）明显高于L9981（657±57）和L9981-pLXSN（705±75）细胞系（P<0.001）；（3）L9981-nm23-H1细胞质中β-连环蛋白表达量（3 649±118）明显高于L9981（1 401±31）和L9981-pLXSN（1 350±55）细胞系（P<0.001）；（4）L9981-nm23-H1（2 945±68）、L9981（2 604±23）和L9981-pLXSN（2 652±53）细胞核中β-连环蛋白表达量差异无统计学意义（P>0.05）；（5）L9981与L9981-pLXSN细胞质和细胞核中GSK-3β或β-连环蛋白表达量差异无统计学意义（P>0.05）。
（1）nm23-H1基因可显著上调L9981-nm23-H1细胞中细胞质和细胞核中GSK-3β的表达以及细胞质中β-连环蛋白的表达，但不能诱导β-连环蛋白在细胞核中积聚。（2）调节GSK-3β和β-连环蛋白表达以及靶向阻断Wnt信号通路可能是nm23-H1在“肺癌转移抑制级联反应”中发挥重要作用的分子机制之一。
