诱导性多能干细胞相关基因OCT4过表达对Hep2细胞增殖及迁移的影响

Overexpression of Induced Pluripotent Stem Cell Related Gene OCT4 on Hep2 Cell Proliferation and Migration Variability

目的应用基因转染技术将诱导性多能干细胞(iPS)相关基因OCT4过表达于喉癌Hep2细胞,观察其对Hep2细胞增殖力和侵袭迁移力的影响。方法构建重组质粒真核表达载体pcD-NA3.1-OCT4,将重组表达质粒用脂质体LipofactaminTM2000转染人喉癌Hep2细胞,通过G418筛选,RT-PCR及Western-blot印迹法鉴定,进而筛选出稳定表达pcDNA3.1-OCT4的Hep2细胞系;MTT实验检测细胞的活力,Transwell法检测细胞的侵袭和迁移。实验分为三组,Hep2-pcDNA3.1-OCT4组、Hep2-pcDNA3.1组及Hep2空白对照组。结果成功将OCT4基因转染入Hep2细胞中,Hep2-pcD-NA3.1-OCT4组可检测到目的基因在核酸、蛋白水平的高表达。转染后的细胞增殖、迁移加快,但细胞形态基本无变化。结论 iPS相关基因OCT4的转染能加快肿瘤细胞的生长和迁移,使Hep2细胞具有干细胞属性。

Objective To observe overexpression of induced pluripotent stem cell（iPS）related gene OCT4 in Hep2 cell and its influence on cell proliferation and migration with gene transfection method.Methods Recombinant plasmid eukaryotic expression vector pcDNA3.1-OCT4 was constructed and transfected into Hep2 cells,then,cells were screened by G418 and detected by qualitative PCR,real time PCR and Western blot.Cell proliferation inhibition was measured by MTT assay,cell migration was observed by Transwell assay.The cells were divided into 3 groups,group Hep2-pcDNA3.1-OCT4,group Hep2-pcDNA3.1 and Hep2 blank control group.Results pcDNA3.1-OCT4 was transfected into Hep2 cells,and the expression of OCT4 was detected in nucleic acids and protein.MTT results showed that Hep2-OCT4 cells grew faster than Hep2 cells.Cell migration results showed the number of cell migration in Hep2-OCT4 cells were higher than the number of Hep2 cells.Conclusion iPS related gene transfection of OCT4 could make Hep2 cells get stem cell properties,accelerate the growth of tumor cells and migration.