Effects Of Overexpression Of VCAM-1 On The Migration In Vitro Of Mesenchymal Stem Cells

OBJECTIVE:To investigate the effect of vascular cell adhesion molecule-1(VCAM-1) on the morphology.growth characteristic.migration in vitro of the muririe mesenchymal stem cells(MSC) and its mechanisms.METHODS:The morphology of murine MSC(C3H10T1/2), VCAM-1 transfected MSC (C3H10T1/2-MIGR1-VCAM-1) and empty vector-transfected MSC(C3H10T1/2-MIGR1) were observed by Giemsa staining. The growth characteristic of murine MSC, VCAM-1 transfected MSC and empty vector-transfected MSC were studied by CCK8. Extracting the protein from murine MSC, VCAM-1 transfected MSC and empty vector-transfected MSC, and using western blot to research their MAPK signal pathways. The migration ability of murine MSC,VCAM-1 transfected MSC and empty vector-transfected MSC were assessed by using the transwell culture system in vitro. The cells were seeded on the membrane with 8 ?m aperture and the fetal bovine serum was used as the chemotactic agent to induce MSC migration. The transmigrated cells were detected with crystal purple as well as DAPI staining for 8h and 12h respectively. The absolute transmigrated cell numbers and the migration rates of MSC were evaluated in each group. Furthermore, the specific chemical inhibitors of MAPK pathway (SB203580, PD98059 and JNK inhibitor ?) were added to the transwell system and the alteration of the MSC migration ability were evaluated at 12h.RESULTS:There were no significant different about morphology in murine MSC (C3H1 0T1/2) and VCAM-1 transfected MSC (C3H10T1/2-MIGR1-VCAM-1). Overexpression VCAM-1 in MSC can inhibit their growth. After incubation with fetal bovine serum for 8h and 12h, the migration ability of VCAM-1-transfected MSC was significantly increased. Both absolute cell number and migration rate of C3H10T1/2-MIGR1-VCAM-1 were obviously up-regulated, but there was no significantly difference between C3H10T1/2-MIGR1 and C3H10T1/2 group. Morever, the migration effect of C3H10T1/2-MIGR1-VCAM-1 was partially suppressed by addition of JNK inhibitor ?, which the transmigrated cell numbers and the migration rate were decreased. However, the VCAM-1 promoting migration of MSC was not suppressed by the inhibitors for ERK/MAPK and p38/MAPK pathway.CONCLUSION:Overexpression of VCAM-1 had no effect on cell morphology. Compared with C3H10T1/2-MIGR1 and C3H10T1/2,the proliferation rate of C3H10T1 /2-MIGR1-VCAM-1 was the lowest. VCAM-1 can increase mouse MSC migration in vitro by JNK/MAPK pathway.