Objective: To investigate the effects and mechanisms of zinc finger E-box binding homeobox transcription factor-2 ( ZEB2) on the proliferation, colony formation, migration, and invasion abilities and the epithelial-mesenchymal transition (EMT) of PANC-1 cells, a human pancreatic cancer cell line.
Methods: Data on the expression of ZEB2 in pancreatic cancer tissues and paracancerous tissues from The Cancer Genome Atlas (TCGA) database were analyzed. PANC-1 pancreatic cancer cells were divided into si-NC group, si- ZEB2 group, pcDNA3.1 group, and pcDNA3.1- ZEB2 group. qRT-PCR and Western blot were conducted to confirm the effectiveness of ZEB2 knockdown or overexpression. CCK-8, colony formation, wound healing, and Transwell assays were conducted to examine the effects of ZEB2 on the proliferation, colony formation, migration, and invasion of PANC-1 cells. qRT-PCR and immunofluorescence assays were performed to examine the expression of E-cadherin and vimentin, the EMT markers, in the cells. Prediction of proteins interacting with ZEB2 was made through the STRING database.
Results: TCGA database analysis showed that the expression level of ZEB2 in pancreatic cancer tissues was significantly higher than that in adjacent tissues ( P<0.05). Compared with those of cells in the control group, the proliferation, colony formation, migration, and invasion of cells in the si- ZEB2 group were decreased ( P<0.05). Compared with those of cells in the pcDNA3.1 group, the proliferation, colony formation, migration and invasion of cells in the pcDNA3.1- ZEB2 group were increased (all P<0.05). According to the results of qRT-PCR and immunofluorescence assays, compared with those of the si-NC group, the expression of E-cadherin mRNA, an epithelial marker, in the si- ZEB2 group increased, while the expression of vimentin mRNA, an mesenchymal marker, and the protein decreased. Compared with those of the pcDNA3.1 group, the expression of E-cadherin mRNA in the PANC-1 cells of the pcDNA3.1- ZEB2 group decreased, while the expression of vimentin mRNA and the protein increased (all P<0.05). Analysis with the STRING database predicted that 10 proteins had close interaction with ZEB2.
Conclusion: Overexpression of ZEB2 promotes the migration, invasion, and the EMT process of PANC-1 pancreatic cancer cells.
目的: 研究锌指E-box结合同源异型盒2基因（zinc finger E-box binding homeobox transcription factor-2, ZEB2）对胰腺癌PANC-1细胞增殖、集落形成、迁移和侵袭能力及上皮-间质转化（epithelial-mesenchymal transition, EMT）过程的影响。
方法: 分析癌症基因组图谱（The Cancer Genome Atlas, TCGA）数据库中胰腺癌组织和癌旁组织中ZEB2的表达。将胰腺癌PANC-1细胞分为si-NC组、si-ZEB2组、pcDNA3.1组和pcDNA3.1-ZEB2组，采用qRT-PCR技术和Western blot法验证ZEB2敲降或过表达的有效性。CCK-8实验、集落形成实验、划痕实验和Transwell实验分别检测ZEB2对PANC-1细胞的增殖、集落形成、迁移和侵袭的影响。利用qRT-PCR技术和免疫荧光实验检测细胞中EMT标志物E-cadherin、vimentin的表达水平。通过STRING网站预测与ZEB2具有相互作用的蛋白。
结果: TCGA数据库分析显示，与癌旁组织相比，胰腺癌组织中ZEB2表达水平明显升高（P<0.05）。与si-NC组相比，si-ZEB2组PANC-1细胞的增殖、集落形成、迁移、侵袭能力减弱；与pcDNA3.1组相比，pcDNA3.1-ZEB2组PANC-1细胞的增殖、集落形成、迁移、侵袭能力增强（均P<0.05）。qRT-PCR技术和免疫荧光实验结果提示，与si-NC组相比，si-ZEB2组PANC-1细胞的上皮标志物E-cadherin mRNA表达升高，而间质标志物vimentin mRNA和蛋白表达降低；与pcDNA3.1组相比，pcDNA3.1-ZEB2组PANC-1细胞中上皮标志物E-cadherin mRNA表达减少，间质标志物vimentin mRNA和蛋白表达增加（均P<0.05）。STRING网站预测出10个蛋白与ZEB2作用密切。
结论: 过表达ZEB2能够促进胰腺癌PANC-1细胞的迁移、侵袭和EMT进程。
Keywords: Invasion; Migration; PANC-1 cell; Pancreatic cancer; Proliferation; ZEB2.
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