Study On The Adhesive Function And Mechanism Of E-Cadherin In Endothelial Progenitor Cells And Mesenchymal Stem Cells

Objective: To explore the adhesive function and mechanism of E-cadherin(E-cad) in endothelial progenitor cells(EPCs) and mesenchymal stem cells(MSCs).Methods 1. C57BL/6J(WT) mice as a cells source. EPCs and MSCs were isolated and cultured from the bone marrow cavity. 2. Mito Tracker Red and DAPI were used to label the EPCs and MSCs respectively. 3. Immunofluorescence was used to examine the E-cad expression of the two cell lines. 4. Cell adhesion assay was used to observe the adhesion of EPCs and MSCs in vitro. 5. E-cad Si-RNA was used to knock down the expression of E-cad and further to observe the adhesion phenomenon of MSCs and EPCs. 6. RT-PCR and Western-blot were used to identify the transfection efficiency of Si-RNA. 7. The adhesion of EPCs and MSCs on the angiogenic Matrigel was further detected after successful transfection. 8. ELISA analysis of E-cad expression in the culture medium. 9. RT-PCR and Western blot were used to determine the E-cad/β-catenin m RNA and protein expression after the mutual effects between MSCs and EPCs. 10. After the addition of β-catenin-specific inhibitor XAV939, the adhesion of EPCs and MSCs on the angiogenic Matrigel was further detected.Results 1. E-cad expression was significantly increased when co-cultured with MSCs and EPCs in vitro. 2. E-cad Si-RNA was successfully transfected with MSCs and EPCs, and the expression of E-cad m RNA and protein were significantly down regulated. 3. The adhesion of MSCs and EPCs was significantly inhibited after successful transfection of Si-RNA. 4. Adhesion and tube formation of EPCs and MSCs were significantly inhibited in the angiogenic Matrigel after E-cad expression of these two cell lines was silenced. 5. Both MSCs and EPCs can secrete E-cad. 6. The E-cad of EPCs may regulate the adhesion between EPCs and MSCs by activating and up-regulating β-catenin in MSCs. 7. β-Catenin-regulated signaling pathway of MSC and EPC adhesion.Conclusions: E-cadherin-mediated contact of endothelial progenitor cells with mesenchymal stem cells through β-catenin signaling.