Effect of different concentrations of Netrin-1 on proliferation and apoptosis of TEV-1 cells

Objective To explore the role of different dose of Netrin-1 on regulating the proliferation and apoptosis of human trophoblast TEV-1 cells. Methods TEV-1 cells were treated with 0,10,100 and 1 000 ng/ml Netrin-1 for 24 h.Cell viability was measured by MTT assay;cell apoptosis were assessed by flow cytometry;the expression of Netrin-1 in TEV-1cells was observed by indirect cellular immunofluorescence;the mRNA and protein of Netrin-1 expression in TEV-1 cells was performed by real-time PCR and Western blot. Results Immediately after 24 hour of treatment with Netrin-1,the proliferation of TEV-1 cells was independent of Netrin-1,meanwhile,cell apoptosis decreased with the accrescence of concentration.The apoptosis rate significantly decreased to (6.63±0.07)% at 1 000 ng/ml Netrin-1 than (14.44±0.85)% without Netrin-1(P<0.05).Immunoreactivity of Netrin-1 was observed in cytoplasm of trophoblast.The expression amount of Netrin-1 protein in TEV-1 cells was independent of the accrescence of concentration by using Western blot. Conclusion Netrin-1 can reduce cell apoptosis in TEV-1 cells,but the expression level of Netrin-1 may not involve in it.