Kim Eunjin, Kang Jeongwoo, Cho Minchul, Lee Sojung, Seo Eunhee, Choi Heesook, Kim Yumi, Kim Junghee, Kang Kum Yong, Kim Kwang Pyo, Han Jaeyong, Sheen Yhunyhong, Yum Young Na, Park Sue-Nie, Yoon Do-Young
Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Korea.
Mol Med Rep. 2009 Jan-Feb;2(1):129-37. doi: 10.3892/mmr_00000073.
The E6 and E7 oncoproteins of human papilloma virus (HPV) type 16 have been known to cooperatively induce the immortalization and transformation of primary keratinocytes. We established an E7 transgenic mouse model to screen HPV-related biomakers using the omics approach. The methods used to identify HPV-modulated factors were genomics analysis by microarray using the Affymetrix 430 2.0 array to screen E7-modulated genes, and proteomics analysis using nano-LC-ESI-MS/MS to screen E7-modulated proteins with the lung tissue of E7 transgenic mice. According to omics data, cyclin B1, cyclin E2, topoisomerase IIα, calnexin, activated leukocyte cell adhesion molecule CD166, actinin α1, diaphorase 1, gelsolin, platelet glycoprotein, and annexin A2 and A4 were up-regulated in the E7-Tg mice, while proteoglycan 4, sarcolipin, titin, vimentin, drep 1, troponin and cofilin-1 were down-regulated. We further confirmed the significance of differences between the expression levels of the selected factors in E7-Tg and non-Tg mice by real-time PCR. Genes related to cancer cell adhesion, cell cycle and migration, proliferation and apoptosis, as well as to the intermediate filament network and to endoplasmic reticulum proteins, were selected. Taken together, the results suggest that the E7 oncogene modulates the expression levels of cell cycle-related (cyclin B1, cyclin E2) and cell adhesion- and migration-related (actinin α1, CD166) factors, which may play important roles in cellular transformation in cancer. In addition, the solubilization of the rigid intermediate filament network by specific proteolysis mediated via up-regulating gelsolin and down-regulating cofilin-1, as well as increased levels of endoplasmic reticulum protein calnexin with chaperone functions, might also be involved in E7-lung epithelial cells.
已知人乳头瘤病毒16型（HPV-16）的E6和E7癌蛋白可协同诱导原代角质形成细胞永生化和转化。我们建立了一个E7转基因小鼠模型，使用组学方法筛选HPV相关生物标志物。用于鉴定HPV调节因子的方法包括：使用Affymetrix 430 2.0芯片进行微阵列基因组分析，以筛选E7调节基因；使用纳米液相色谱-电喷雾串联质谱（nano-LC-ESI-MS/MS）进行蛋白质组学分析，以筛选E7转基因小鼠肺组织中的E7调节蛋白。根据组学数据，在E7转基因小鼠中，细胞周期蛋白B1、细胞周期蛋白E2、拓扑异构酶IIα、钙连接蛋白、活化白细胞细胞粘附分子CD166、辅肌动蛋白α1、黄递酶1、凝溶胶蛋白、血小板糖蛋白以及膜联蛋白A2和A4上调，而蛋白聚糖4、肌质蛋白、肌联蛋白、波形蛋白、drep 1、肌钙蛋白和丝切蛋白-1下调。我们通过实时PCR进一步证实了E7转基因小鼠和非转基因小鼠中所选因子表达水平差异的显著性。选择了与癌细胞粘附、细胞周期和迁移、增殖和凋亡以及中间丝网络和内质网蛋白相关的基因。综上所述，结果表明E7癌基因调节细胞周期相关（细胞周期蛋白B1、细胞周期蛋白E2）以及细胞粘附和迁移相关（辅肌动蛋白α1、CD166）因子的表达水平，这可能在癌症细胞转化中起重要作用。此外，通过上调凝溶胶蛋白和下调丝切蛋白-1介导的特异性蛋白水解作用使刚性中间丝网络溶解，以及具有伴侣功能的内质网蛋白钙连接蛋白水平升高，也可能与E7-肺上皮细胞有关。
