Studies On The Mechanism Of Apoptosis In U251 Cells By Interfering BDNF Expression By SiRNA

ObjectiveTo discuss the effect of si RNA specificly for BDNF on U251 cells apoptosis and its related mechanism.MethodsHuman gliomas cell line U251 were cultured in vitro and divided into three groups, group A as control group, group B as non-silencing-si RNA group, group C as the BDNF-si RNA group. The logarithmic phase of glioma U251 cells used in this study. Expression of BDNF, AKt and p AKt in three groups was examined by Western blot and BDNF secretion level was evaluated by ELISA. Cell apoptosis in each group were examined by flow cytometry instrument method.ResultsThe U251 cells were transiently transfected by non-silencing-si RNA and BDNF-si RNA. Western blot results showed that compared with group A and group B, protein expression of BDNF, p AKt in group C was decreased, the difference was statistically significant(P<0.05); compared with A, B, there was no statistically significant difference. ELISA results showed that BDNF concentration in the supernatant of group C was(70.20±3.05)pg/ml, which lower than group A, the difference was statistically significant(P<0.05); No obvious difference was found between A and B. The cell apoptosis rate of group C was(19.62±2.50)%, which was higher than group A(1.63±0.61)% and group B(1.78±0.94)%, the difference was statistically significant(P<0.05); Compared with A and B, there was no statistically significant difference.ConclusionsThe human brain derived neurotrophic factor(BDNF) have the ability to facilitate the glioma cells survive and antiapoptotic. When U251 cells were cultured in vitro, specific BDNF-si RNA can significantly reduce the expression of BDNF, p AKt, meanwhile, when interference expression of BDNF, the apoptosis of U251 cells was Increased, we suspect that this is closely related to the Trk B-Akt pathway, Its specific mechanism needs follow-up of further research.