The Construction of E-Cadherin Lentivirus Vectors and the Experimental Study of E-Cadherin on Neural Stem Cell Biological Behavior

Using neural stem cell (NSC) transplantation to reconstruct neural circuit is one of the important approaches to treat spinal cord injury. E-cadherin is considered as one the most important factors that can regulate microenvironment of NSCs, and further to improve the treatment. In this study, based on the sequence of E-cadherin reported by Genbank, we designed the relevant RNAi sequence. The designed siRNA fragments were inserted into the packaging plasmid PHR-SIN-CSGW-H1a (PHR) to construct E-cadherin RNAi lentivirus vector. By linking E-cadherin and pcDNA3.1 plasmid, E-cadherin over-expression lentivirus vector was constructed. The expression of E-cadherin and N-cadherin in NSCs was detected with Western Blot and Real Time PCR. Using these E-cadherin lentivirus vectors, the effect of E-cadherin on NSC proliferation and migration was examined. Our results indicated that after transfection, E-cadherin expression from NSCs was increased with over expression and decreased with RNAi. For N-cadherin, it is just opposite. Overexpression of E-cadherin can remarkably increase NSC proliferation rate. However, RNAi did not alter NSC proliferation rate. Meanwhile, over expression of E-cadherin lowered NSC migration ability. On the other hand, down regulation of E-cadherin expression with RNAi lentivirus vector enhanced NSC migration ability. These vitro experiment results have set up a good basis for the subsequent study of using NSCs transplantation to repair spinal cord injury.