摘要
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 metalloproteinase2,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RHOCmRNA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。
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(P0.05).The expression level of TOB1 mRNA was positively correlated with the expression levels of PTEN,BRMS1 and NME1 mRNAs(P0.05).The abilities of migration and invasion of the A549/TOB1 cells were obviously decreased as compared with those of the A549 cells(P0.05).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(P0.05).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(P0.05).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.
出处
《肿瘤》
CAS
CSCD
北大核心
2012年第2期85-91,共7页
Tumor
基金
江苏省高校自然科学基础研究面上项目(编号:08KJB310012)