TOB1高表达抑制肺癌A549细胞体外迁移和侵袭

目的:探讨TOB1 (transducer of ErbB 2,1)高表达对人肺癌A549细胞体外迁移和侵袭的影响.方法:应用RT-PCR法检测人支气管上皮细胞HBE和10种肺癌细胞中TOB1、PTEN (phosphatase and tensin homolog deleted on chromosome ten)、乳腺癌转移抑制基因1(breast cancer metastasis suppressor 1,BRMS1)、RHOC(ras homolog gene family,member C)和NME1 (non-metastatic cells 1)mRNA的表达水平；将重组表达载体质粒pcDNA3.0-TOB1和“空载体”质粒pcDNA3.0转染到A549细胞中,建立TOB1高表达的A549/TOB1细胞和对照组A549/PC3细胞,应用体外划痕和Transwell实验检测细胞的体外迁移和侵袭能力,RT-PCR法检测细胞中PTEN、BRMS1、RHOC和NME1 mRNA的表达,蛋白质印迹法检测细胞中α-N-catenin、β-catenin、γ-catenin、基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)和MMP9蛋白的表达.结果:10种肺癌细胞中TOB1 mRNA的表达水平明显低于HBE细胞(P＜0.05),不同细胞株中TOB1 mRNA与PTEN、BRMS1和NME1 mRNA的表达水平间呈正相关(P＜0.05).A549/TOB1细胞的体外迁移和侵袭能力明显低于A549细胞(P＜0.05).A549/TOB1细胞中PTEN、BRMS1和NME1 mRNA的表达水平明显高于A549细胞(P＜0.05),而RHOC mRNA的表达水平明显低于A549细胞(P＜0.05); A549/TOB1细胞中α-N-catenin、β-catenin、γ-catenin、MMP2和MMP9蛋白的表达水平明显低于A549细胞(P＜0.05).结论:外源性TOB1高表达可以明显抑制肺癌A549细胞的体外转移和侵袭,其机制可能与TOB1调控PTEN、BRMS1、RHOC和NME1的表达,进一步下调α-N-catenin、β-catenin、γ-catenin、MMP2和MMP9蛋白的表达水平有关.

Over-expression of transducer of ErbB2,1 inhibits migration and invasion of human lung cancer cell line A549 in vitro

Objective:To investigate the effect of over-expression of transducer of ErbB2,1(TOB1) on migration and invasion of human lung adenocarcinoma cell line A549.Methods:The expression levels of TOB1,phosphatase and tensin homolog deleted on chromosome ten(PTEN),breast cancer metastasis suppressor 1(BRMS1),ras homolog gene family,member C(RHOC) and non-metastatic cells 1(NME1) mRNAs in human bronchial epithelial(HBE) cells and ten types of human lung cancer cells were detected by RT-PCR.The recombinant plasmid pcDNA3.0-TOB1 and the empty vector pcDNA3.0 were transfected into A549 cells,respectively.The A549 cells over-expressing TOB1 protein(A549/TOB1 cells) and the A549/PC3 cells(as the control cells) were established.The abilities of migration and invasion of A549/TOB1 cells were determined by wound healing and Transwell assay.The expression levels of PTEN,BRMS1,RHOC and NME1 mRNAs were detected by RT-PCR,and the expression levels of α-N-catenin,β-catenin,γ-catenin,matrix metalloproteinase 2(MMP2) and MMP9 were detected by Western blotting.Results:The expression levels of TOB1 mRNA in ten types of lung cancer cells were lower than that in HBE cells(P<0.05).The expression level of TOB1 mRNA was positively correlated with the expression levels of PTEN,BRMS1 and NME1 mRNAs(P<0.05).The abilities of migration and invasion of the A549/TOB1 cells were obviously decreased as compared with those of the A549 cells(P<0.05).The expression levels of PTEN,BRMS1 and NME1 mRNAs were higher whereas the expression level of RHOC mRNA was lower in the A549/TOB1 cells as compared with those in the A549 cells(P<0.05).The expression levels of α-N-catenin,β-catenin,γ-catenin,MMP2 and MMP9 proteins in the A549/TOB1 cells were lower than those in the A549 cells(P<0.05).Conclusion:Exogenous TOB1 over-expression can inhibit the abilities of migration and invasion of A549 human lung cancer cells in vitro.This effect may be associated with the down-regulation of α-N-catenin,β-cateninm γ-catenin,MMP2 and MMP9 expressions,and which was induced through the regulation of PTEN,BRMS1,RHOC and NME1 expressions by TOB1.