E-cadherin基因稳定转染小鼠胚胎干细胞及其对分化细胞黏附能力的影响

E-cadherin stable transfection into mouse ES cells and its effects on adhesion ability of differentiated cells in vitro

目的转染上皮型钙黏蛋白（E-cadherin）基因入小鼠胚胎干（Es）细胞，观察其对分化细胞间黏附能力变化的影响。方法构建E-cadherin的真核表达载体pEGFP-E-cadherin，转染小鼠ES细胞并进行体外分化。利用逆转录一聚合酶链反应（RT-PCR）等检测E-cadherin在分化系统中的动态表达，同时观察分化细胞问黏附能力的变化情况。结果基因转染的ES细胞分化后第1～17天稳定表达E-cadherin，而普通ES细胞分化后则表达逐渐降低。稳定表达E-cadherin的分化细胞问黏附能力明显增强并在19d时仍能保持致密的细胞连接和多层生长状态，普通ES细胞则由拟胚体（EB）结构逐渐分化为松散的细胞群落。结论稳定表达E—cadherin的ES细胞分化后，细胞间黏附能力明显增强并保持多层细胞生长状态，这对于将ES细胞分化研究提升到组织水平具有积极的意义。

Objective To stably transfect E-cadherin into embryonic stem ceils （ESCs） ,and observe the impact of E-eadherin on adhesion ability of differentiated cells. Methods BALB/c mouse ESCs were infected with recombinant plasmid EGFP-E-cadherin. E-cadherin dynamic expression in E-cadherin- ESCs differentiation system was detected by using RT-PCR and ICC. The appearances of cell-cell adhesion in the E-cadherin-ESCs differentiating system was observed dynamically under a microscope. Results The exogenous E-cadherin gene was successfully transferred into BALB/c mouse ESCs and expressed stably. In E-cadherin-ESCs differentiation system,E-cadherin-mRNA was expressed stably during the differentiating period from day 1 to day 17. But in un-infected ESCs differentiation system, E-cadherin-mRNA was not expressed till on day 5 and expressed weakly along with the differentiating time. At last,the E-cadherin-ESCs formed pronounced cell aggregates and cuboidal morphology, whereas un-infected ESCs remained more spread out and corded in morphology. Conclusion Transferring exogenous E-cadherin gene into mouse ESCs could enhance the cell adhesion and maintain multi-laver form in ESCs differentiation system.