Yin Xue-Jun, Xu Jian-Ning, Zou Chang-Qi, He Feng-Sheng, Fang Fu-De
Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Beijing 100005, China.
Biomed Environ Sci. 2004 Dec;17(4):432-41.
To define the differences in gene expression patterns between glycidyl methacrylate (GMA)-transformed human lung fibroblast cells (2BS cells) and controls.
The mRNA differential display polymerase chain reaction (DD-PCR) technique was used. cDNAs were synthesized by reverse transcription and amplified by PCR using 30 primer combinations. After being screened by dot blot analysis, differentially expressed cDNAs were cloned, sequenced and confirmed by Northern blot analysis.
Eighteen differentially expressed cDNAs were cloned and sequenced, of which 17 were highly homologous to known genes (homology = 89%-100%) and one was an unknown gene. Northern blot analysis confirmed that eight genes encoding human zinc finger protein 217 (ZNF217), mixed-lineage kinase 3 (MLK-3), ribosomal protein (RP) L15, RPL41, RPS 16, TBX3, stanniocalcin 2 (STC2) and mouse ubiquitin conjugating enzyme (UBC), respectively, were up-regulated, and three genes including human transforming growth factor beta inducible gene (Betaig-h3), alpha-1,2-mannosidase 1A2 (MAN 1A2) gene and an unknown gene were down-regulated in the GMA-transformed cells.
Analysis of the potential function of these genes suggest that they may be possibly linked to a variety of cellular processes such as transcription, signal transduction, protein synthesis and growth, and that their differential expression could contribute to the GMA-induced neoplastic transformation.
确定甲基丙烯酸缩水甘油酯（GMA）转化的人肺成纤维细胞（2BS细胞）与对照细胞之间基因表达模式的差异。
采用mRNA差异显示聚合酶链反应（DD-PCR）技术。通过逆转录合成cDNA，并使用30种引物组合通过PCR进行扩增。经斑点印迹分析筛选后，对差异表达的cDNA进行克隆、测序，并通过Northern印迹分析进行验证。
克隆并测序了18个差异表达的cDNA，其中17个与已知基因高度同源（同源性=89%-100%），1个为未知基因。Northern印迹分析证实，分别编码人锌指蛋白217（ZNF217）、混合谱系激酶3（MLK-3）、核糖体蛋白（RP）L15、RPL41、RPS 16、TBX3、鲽钙蛋白2（STC2）和小鼠泛素缀合酶（UBC）的8个基因上调，而包括人转化生长因子β诱导基因（Betaig-h3）、α-1,2-甘露糖苷酶1A2（MAN 1A2）基因和1个未知基因在内的3个基因在GMA转化细胞中下调。
对这些基因潜在功能的分析表明，它们可能与转录、信号转导、蛋白质合成和生长等多种细胞过程相关，且它们的差异表达可能促成了GMA诱导的肿瘤转化。
