Li Jin-zhong, Pan Hong-ya, Zheng Jia-wei, Zhou Xiao-jian, Zhang Ping, Chen Wan-tao, Zhang Zhi-yuan
Department of Oral and Maxillofacial Surgery, College of Stomatology, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Chin Med J (Engl). 2008 Oct 5;121(19):1882-90.
The present study was designed to examine and analyze the global gene expression changes during the tumorigenesis of a human immortalized oral epithelial cell line, and search for the possible genes that may play a role in the carcinogenesis of oral cancer associated with benzo (a) pyrene.
The human immortalized oral epithelial cells, which have been established through transfection of E6/E7 genes of human papillomavirus type 16 and proved to be non-tumorigenic in nude mice, were treated with benzo (a) pyrene. Tumorigenicity of the treated cells were examined through nude mice subcutaneous injection. The global gene expression profiles of immortalized cells and the tumorigenic cells were acquired through hybridization of a microarray of Affymetrix U133 plus 2.0. The data were analyzed using Spring 7.0 software and treated statistically using one-way analysis of variance (ANOVA). The differentially expressed genes were classified using a Venn diagram and annotated with gene ontology. Several highlighted genes were validated in cells using a real-time polymerase chain reaction.
There were 883 differentially expressed genes during the tumorigenesis and most of them changed expression in the early stage of tumorigenesis. These genes mainly involved in macromolecule metabolism and signal transduction, possessed the molecular function of transition metal ion binding, nucleotide binding and kinase activity; their protein products were mainly integral to membranes or localized in the nucleus and cytoskeleton. The expression patterns of IGFBP3, S100A8, MAP2K, KRT6B, GDF15, MET were validated in cells using a real-time polymerase chain reaction; the expression of IGFBP3 was further validated in clinical oral cancer specimens.
This study provides the global transcription profiling associated with the tumorigenesis of oral epithelial cells exposed to benzo (a) pyrene; IGFBP3 may play a potential role in the initiation of oral cancer related to benzo (a) pyrene exposure.
本研究旨在检测和分析人永生化口腔上皮细胞系肿瘤发生过程中的全基因组表达变化，并寻找可能在苯并（a）芘相关口腔癌致癌过程中发挥作用的基因。
将通过转染人乳头瘤病毒16型E6/E7基因建立且已证实在裸鼠中无致瘤性的人永生化口腔上皮细胞用苯并（a）芘处理。通过裸鼠皮下注射检测处理后细胞的致瘤性。通过Affymetrix U133 plus 2.0芯片杂交获取永生化细胞和致瘤细胞的全基因组表达谱。使用Spring 7.0软件分析数据，并采用单因素方差分析（ANOVA）进行统计学处理。使用维恩图对差异表达基因进行分类并用基因本体进行注释。在细胞中使用实时聚合酶链反应对几个突出的基因进行验证。
肿瘤发生过程中有883个差异表达基因，其中大部分在肿瘤发生早期表达发生变化。这些基因主要参与大分子代谢和信号转导，具有过渡金属离子结合、核苷酸结合和激酶活性的分子功能；其蛋白质产物主要整合到膜中或定位于细胞核和细胞骨架。使用实时聚合酶链反应在细胞中验证了IGFBP3、S100A8、MAP2K、KRT6B、GDF15、MET的表达模式；在临床口腔癌标本中进一步验证了IGFBP3的表达。
本研究提供了与暴露于苯并（a）芘的口腔上皮细胞肿瘤发生相关的全转录组图谱；IGFBP3可能在与苯并（a）芘暴露相关的口腔癌发生起始中发挥潜在作用。
